Objective:To explore the clinical phenotypic features and genetic variation characteristics of children with epilepsy-aphasia spectrum due to GRIN2A gene variants confirmed by second-generation sequencing. Methods:The clinical data of 5 children with epilepsy-aphasia spectrum with epileptic onset diagnosed in the Department of Neurology, Children′s Hospital Affiliated to Zhengzhou University, from February 2019 to November 2022 were retrospectively analyzed. Whole-exome genome sequencing of the probands using a second-generation sequencing method confirmed that all 5 cases were children with the GRIN2A gene variant. The characteristics of the GRIN2A gene variants were analyzed. Results:Among the 5 children diagnosed with epileptic aphasia spectrum due to GRIN2A gene variants, the male-to-female ratio was 4∶1, and the age range of onset was 1.5-4.4 years. The clinical phenotype included seizures in all cases, language and intellectual developmental deficits in 4 cases, and attention deficit hyperactivity disorder in 3 cases. The seizures were manifested as focal seizures or secondary generalized seizures, and were effectively controlled with antiepileptic drugs. Among the 5 children, gene variant of case 1 was originated from a paternal heterozygous variant, and cases 2-5 had de novo variants, which were c.2107C>T (p.Gln703 *) nonsense variant, c.2284G>A (p.Gly762Arg) missense variant, c.2197del (p.Ala733Glnfs *3) shifted coding variant, c.2511G>A (p.Trp837 *) nonsense variant, and c.1651+1G>C shear site variant, respectively. None of the 5 loci were reported in the literature. Conclusions:Epilepsy-aphasia spectrum is an epilepsy syndrome with a complex onset, and may have different phenotypes at different genetic variant loci, with focal seizures or secondary generalized seizures, which can be effectively controlled with anti-seizure medication. The GRIN2A gene variant is the genetic etiology of the epileptic aphasia spectrum.

Objective:To explore the clinical phenotype and genetic etiology for seven children with CHARGE syndrome (CS).Methods:Clinical data of 7 children with CS diagnosed between March 2020 and December 2022 at the Children′s Hospital Affiliated to Zhengzhou University were analyzed. Genomic DNA was extracted from peripheral blood samples from the children and their parents, and subjected to whole exome sequencing. Candidate variants were verified by Sanger sequencing and pathogenicity analysis. This study was approved by the Medical Ethics Committee of the Children′s Hospital Affiliatedto Zhengzhou University (Ethics No. 2024-K-023).Results:The ages of the children had ranged from 1 day after birth to 12 years old, and all of them had shown growth retardation. The reasons for their admission had included postnatal breathing, swallowing and feeding difficulties in five cases. One child was found to have abnormal external genitalia in conjunct with hearing impairment, whilst another child had shown no secondary sexual characteristics during puberty. All of the children were found to harbor CHD7 gene variants, which included 3 nonsense variants, 2 frameshifting variants and 2 missense variants, i. e., c. 6292C>T (p.R2098*), c.2754G>A (p.W918*), c. 469C>T (p.R157*), c. 3308T>A (p.V1103D), c. 7111delC (p.Q2371Kfs), c. 6023delA (p.D2008Vfs) and c. 3565C>T (p.R1189C). All of the variants were de novo in origin. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 3308T>A (p.V1103D) and c. 3565C>T (p.R1189C) variants were rated as likely pathogenic (PS2+ PM2_Supporting+ PP3), whilst the remainders were rated as pathogenic (PVS1+ PS2+ PM2_Supporting). Conclusion:There is strong clinical and genetic heterogeneity in CS. Early genetic testing may facilitate accurate diagnosis. The detection of novel variants has expanded the phenotypic spectrum of CS and the mutational spectrum of the CHD7 gene.

Objective:To analyze the clinical and genetic characteristics of a child featuring Dias-Logan syndrome.Methods:A child with speech disorders and delayed psychomotor development from childhood who was admitted to the Rehabilitation Medicine Department of Children′s Hospital Affiliated to Zhengzhou University in July 2022 was selected as the research subject. Clinical data of the child was collected. Genomic DNA was extracted from peripheral blood samples of the child and his parents. Potential variant was screened by whole exome sequencing, and candidate variant was verified by Sanger sequencing. This study was approved by the Children′s Hospital Affiliated to Zhengzhou University (Ethics No. 2023-K-011).Results:The child has presented with global developmental delay, microcephaly, special facial features and behavioral problems. Genetic testing revealed a de novo variant of the BCL11A gene, namely c. 561_567delACACGCA(p.Q187fs*7), which was classified as pathogenic (PVS1+ PS2+ PM2_Supporting). Conclusion:The heterozygous variant of BCL11A gene probably underlay the Dias-Logan syndrome in this child. Above finding has enriched the phenotypic and mutational spectrum of the BCL11A gene and provides a basis for genetic counseling and clinical decision-making.

Objective:To clarify the genetic diagnosis of two children with ring chromosome 18 and explore their mechanisms and clinical phenotypes.Methods:Two patients treated at the Children′s Hospital of Henan Province respectively in June 2022 and March 2023 were selected as the study subjects. Genetic testing and diagnosis were carried out through copy number variation sequencing (CNV-seq), G-banded chromosomal karyotyping, and whole exome sequencing (WES). This study was approved by the Children′s Hospital of Henan Province (Ethics No. 2023-K-075).Results:Child 1 had mainly manifested developmental delay, white matter hypoplasia, type 1 diabetes mellitus, and micropenis. He was found to have a chromosomal karyotype of 46, XY, r(18)(p11.21q22.1)[40]/46, XY[7], and CNV-seq results showed that he has a 14.86 Mb deletion at 18p11.21p11.32 and a 14.02 Mb deletion at 18q22.1q23. Child 2 had peculiar facial features, delayed white matter myelination, developmental delay, atrial septal defect, severe sensorineural deafness, and congenital laryngeal stridor. He was found to have a chromosomal karyotype of 46, XY, r(18)(p11.2q23). CNV-seq result proved that he had a 14.86 Mb deletion at 18p11.21p11.32 and a 20.74 Mb deletion at 18q21.32q23. WES has failed to detect single nucleotide variants (SNVs) in either child, but revealed a large segmental deletion at chromosome 18 in both of them.Conclusion:Both children were diagnosed with ring chromosome 18 syndrome. The different size of the deletional fragments in the 18q region and mosaicism of ring chromosome 18 in child 1 may underlay the variation in their clinical phenotypes. The type 1 diabetes mellitus and micropenis noted in both children are novel features for ring chromosome 18 syndrome.

Objective:To evaluate the morphological and functional changes of the heart during second trimester fetuses with ventricular septal defect (VSD) using fetal heart quantification (fetal HQ) technology.Methods:A prospective study was conducted from July 2022 to January 2024 at Chengdu Women′s and Children′s Central Hospital, collecting 91 singleton fetuses diagnosed with isolated VSD (VSD group) and 91 normal fetuses matched for gestational age (control group). Fetal HQ technology was used to measure the length and width of the four-chamber view of the fetal heart, obtaining the global sphericity index (GSI). Speckle tracking technology was used to track the endocardial motion trajectories of the left and right ventricles during diastole and systole, obtaining parameters such as left and right ventricular global longitudinal strain (LV-GLS and RV-GLS), end-diastolic diameter (EDD), 24-segment sphericity index (SI), 24-segment fraction of shortening (FS), left ventricular ejection fraction (LV-EF), fraction of area change (FAC), left ventricular stroke volume (LV-SV), and left ventricular cardiac output (LV-CO).The differences between groups were compared, and the correlations between the values of VSD and GSI, GLS, and FAC were evaluated.Results:The EDD of the left ventricular segments 20-23 in the VSD group was lower, while the SI value of the right ventricular segments 1-4 in the VSD group was higher than that in the control group (all P<0.05). There was no statistically significant difference in GSI between the two groups ( P>0.05). LV-GLS in the VSD group was lower than that in the control group ( P<0.05). There was no statistically significant difference in RV-GLS ( P>0.05). Values of LV-FAC, LV-EF, LV-SV, and LV-CO in the VSD group were significantly lower than those in the control group (all P<0.05). The FS value of left ventricular segments 1-10 in the VSD group presented lower, but the FS value of right ventricular segments 7-21 higher compared to controls(all P<0.05). LV-GLS and LV-FAC absolute values were negatively correlated with the size of VSD ( r=-0.309, P=0.004; r=-0.264, P=0.015), while GSI, RV-GLS, and RV-FAC showed no significant correlation with the size of VSD (all P>0.05). Conclusions:The overall sphericity index of second trimester VSD fetuses is normal, but there are changes in the shape of the left ventricular apical segments and the right ventricular basal segments, with the left heart chamber tending to be flatter and the right heart chamber more fusiform. The left ventricular systolic function of VSD fetuses is significantly reduced, the local systolic function of right ventricular increases while the global systolic function shows no significant change. The absolute values of LV-GLS and LV-FAC in VSD fetuses are negatively correlated with the size of VSD.

Objective To compare the lipid-lowering efficacy and safety of fixed-dose combination and free combination of rosuvastatin and ezetimibe in hypercholesterolemia patients who fail to achieve low-density lipoprotein cholesterol(LDL-C)goal with statin monotherapy.Methods A total of 45 hypercholesterolemia patients who switched from statin monotherapy to fixed-dose combination of rosuvastatin and ezetimibe after failing to achieve target LDL-C goal admitted at cardiological departments of First Affiliated Hospital of Sun Yat-sen University,Nanhai Fourth People's Hospital,Foshan First People's Hospital,and Guangdong Provincial People's Hospital between March and June 2024 were enrolled and served as the study group.Another 120 hyper-cholesterolemia patients who treated with free combination of rosuvastatin and ezetimibe were se-lected from Xiamen Regional Health Medical Big Data Platform with propensity score matching and served as control group.The LDL-C level,LDL-C reduction,and changes in TC,HDL-C and TG levels in 4-6 weeks after the medication switch,as well as the safety indicators(AST,ALT,CK,Cre and eGFR)were compared between the two groups.Results In 4-6 weeks after the medication switch,the patients in the study group exhibited a significant decrease in LDL-C level(1.70±0.44 mmol/L vs 2.12±0.87 mmol/L,P＜0.01),obvious LDL-C reduction[(43.17±16.11)％vs(29.14±29.13)％,P＜0.01]when compared to those of the control group.The LDL-C goal attainment rate was significantly higher in the study group than the control group(71.11％vs 45.00％,P=0.003).In addition,there were no statistical differences in the levels of HDL-C and TG and the reductions of HDL-C and TG between the two groups in 4-6 weeks after treatment(P＞0.05).The study group obtained notably lower TC level and TC reduction than the control group in the time(P＜/0.05,P＜0.01).After treatment,no statistical differences were observed between the two groups in terms of AST,ALT,CK,Cre and eGFR(P＞0.05).Conclusion Com-pared to free combination of rosuvastatin and ezetimibe,fixed-dose combination can further reduce LDL-C level in hypercholesterolemia patients who have not achieved LDL-C goal with statin monotherapy,with higher LDL-C goal attainment rate and good safety.

Tongue diagnosis is an important method of TCM diagnosis and treatment.Tongue is the key link of auxiliary diagnosis of tongue feature extraction and processing,and also is the bottleneck of intelligent tongue diagnosis in traditional Chinese medicine.Using image processing,artificial intelligence technology to the tongue as a quantitative and identify characteristics of traditional Chinese medicine,looking for both conforms to the original thinking of TCM,and TCM tongue diagnosis method of accurately,has become a common concern of traditional Chinese medicine and computer field.From the mobile terminal tongue as auxiliary diagnostic system of traditional Chinese medicine tongue acquisition basic attribute,tongue diagnosis and image information building,tongue like features are required for accurate extraction and so on related key technology is analyzed,and build overall architecture,so as to provide technical reference for the tongue like intelligent diagnosis,promote the development of technology of tongue diagnosis in traditional Chinese medicine modernization.

Objective To explore the predictive value of blood lipid dynamics of septic shock for the short-term prognosis.Methods A total of 107 patients with septic shock treated in Dongzhimen Hospital of Beijing University of Traditional Chinese Medicine from March 2019 to July 2023 were selected as the study subjects.They were divided into survival group(n=76)and death group(n=31)based on their 28-day outcomes.Total cholesterol(TC),triacylglycerol(TG),high density lipoprotein cholesterol(HDL-C)and LDL cholesterol(LDL-C)concentrations were tested at disease onset(day 0),day 4±1 and day 7±1.The receiver operating characteristic(ROC)curve was used to analyze the predictive value of lipid dynamics on outcomes in patients with septic shock.Multivariate COX regression analysis was used to analyze factors affecting the prognosis of patients with septic shock.Result The levels of TC and HDL-C at disease onset,as well as TC,HDL-C and LDL-C on day(4±1)and day(7±1)in the survival group were higher than those in death group,and the differences were significant(Z=2.241～5.744,all P＜0.05).The areas under the curve[AUC(95％)]for the prediction of changes in HDL-C and LDL-C in the first week of admission were 0.775(95％CI:0.685～0.850)and 0.646(95％CI:0.547～0.736),with the best cutoff values of 0.16mmol/L and 0.28mmol/L,sensitivity of 70.97％and 64.52％,and specificity of 82.89％and 69.74％,respectively.Multivariate regression analysis showed that HDL-C dynamics was independently associated with poor prognosis(OR=0.141,95％CI:0.044～0.454,P=0.001).Conclusion Monitoring the dynamic changes of blood lipid at the first week of admission could help in predicting short-term outcomes in patients with septic shock.

Objective To study the expression of long non-coding RNA(LncRNA TTN-AS1)TTN-AS1 and squalene epoxidase(SQLE)in rectal cancer,and their correlation with clinicopathological characteristics and prognosis.Methods A total of 90 rectal cancer patients diagnosed and treated in Longquan Hospital of Chengdu University of Traditional Chinese Medicine from January 2018 to January 2020 were selected as the research subjects.Fluorescence quantitative PCR was used to detect the expression of LncRNA TTN-AS1 in tissues.Immunohistochemistry was used to detect the expression of SQLE in tissues.The relationship between LncRNA TTN-AS1,SQLE and clinicopathological characteristics of rectal cancer were compared.K-M curve analysis was used to analyze the impact of LncRNA TTN-AS1 and SQLE on the prognosis of rectal cancer.COX regression analysis was used to analyze factors affecting the prognosis of rectal cancer.Results The relative expression level of Lnc RNA TTN-AS1(3.12±0.45)and the positivity rate of SQLE(71.11％)protein in rectal cancer tissues were higher than those in adjacent tissues(0.91±0.12,8.89％),and the differences were significant(t/x2=45.156,72.593,all P＜0.001).The expression of Lnc RNA TTN-AS1 was positively correlated with SQLE in rectal cancer tissue(r=0.589,P＜0.001).The relative expressions of Lnc RNA TTN-AS1(4.26±0.52,4.10±0.49),SQLE(88.57％,91.43％)in tumor TNM stage Ⅲ and lymph node metastasis were higher than those in tumor TNM stage Ⅰ～Ⅱ(2.39±0.40,60.00％)and tissues without lymph node metastasis(2.50±0.42,58.18％),and the differences were significant(t/x2=8.409～19.211,all P＜0.05).The 3-year survival rates of the LncTTN-AS1 high expression group and low expression group were 50.00％(22/44)and 86.96％(40/46),respectively,and the difference between the curves was significant(Log-rank x=14.205,P=0.001).The 3-year survival rates of the SQLE positive and negative groups were 64.06％(41/64)and 88.46％(23/26),respectively,and the difference between the curves was significant(Log-rank x2=6.291,P=0.012).Lnc RNA TTN-AS1 high expression(HR=2.552,P=0.001),SQLE positive(HR=1.754,P=0.004),tumor TNM stage Ⅲ(HR=1.625,P=0.030),and lymph node metastasis were(HR=2.797,P=0.011)independent risk factors for the prognosis of rectal cancer.Conclusion The increased expression of LncRNA TTN-AS1 and SQLE in rectal cancer tissue are associated with TNM staging and lymph node metastasis,and both are tumor marker for evaluating the prognosis of rectal cancer.

Objective:To investigate the effects and mechanism of metformin on the wound healing of full-thickness skin defects in diabetic rats.Methods:This study was an experimental study. Eighteen 8-week-old male Sprague Dawley rats were divided into control group, diabetes group, and diabetes+metformin group according to complete random grouping method, with 6 rats in each group. The latter two groups of rats were used to create diabetic models, and then four circular full-thickness skin defect wounds with a diameter of 5 mm were made on the back of 18 rats. Metformin F-127 hydrogel was applied only to the wounds of rats in diabetes+metformin group. The wound healing status on post injury day (POD) 7 and 13 was observed and the wound healing rate was calculated. The wound tissue on POD 7 and 13 was collected for hematoxylin-eosin staining to measure the length of re-epithelialized epidermis and calculate the change rates in diameters of epidermal and dermal wounds, for immunohistochemical staining to detect the relative expressions of keratin 10 and proliferating cell nuclear antigen (PCNA), and for Western blotting to detect the protein expressions of keratin 10 and PCNA. The sample size in all the above experiments was 8 except that in the last experiment was 3. The correlations between the relative expressions of keratin 10 and PCNA in wound tissue in three groups of rats and their wound healing rates, and the correlation between the relative expressions of keratin 10 and PCNA in wound tissue were analyzed.Results:On POD 7, the wound healing rates of rats in diabetes group and diabetes+metformin group were 81.48% (77.89%, 85.53%) and 93.04% (92.51%, 94.24%), which were significantly lower than 100% (97.17%, 100%) in control group (with Z values of 2.37 and -3.36, respectively, P<0.05); the wound healing rate of rats in diabetes+metformin group was significantly higher than that in diabetes group ( Z=3.45, P<0.05). On POD 13, the wound healing rates of rats in control group and diabetes+metformin group were both 100% (100%, 100%), which were significantly higher than 94.47% (90.68%, 99.82%) in diabetes group (with Z values of 2.90 and -2.90, respectively, P<0.05). On POD 7, the change rates in epidermal wound diameter of rats in control group and diabetes+metformin group were significantly higher than that in diabetes group (with Z values of 3.36 and -2.74, respectively, P<0.05). The change rates in dermal wound diameter of rats in the three groups were similar on POD 7 and 13 ( P>0.05). The lengths of re-epithelialized epidermis of rats in control group and diabetes+metformin group on POD 13 were significantly longer than that in diabetes group (with Z values of 3.34 and -2.64, respectively, P<0.05). The relative expressions of keratin 10 in wound tissue of rats in diabetes group on POD 7 and 13 were significantly higher than those in control group (with Z values of -3.36 and -3.26, respectively, P<0.05) and diabetes+metformin group (with Z values of 3.36 and 3.15, respectively, P<0.05), and the relative expression of keratin 10 in wound tissue of rats in diabetes+metformin group on POD 7 was significantly lower than that in control group ( Z=3.05, P<0.05); the relative expressions of PCNA in wound tissue of rats in diabetes group on POD 7 and 13 were significantly lower than those in control group (with both Z values of 3.36, P<0.05) and diabetes+metformin group (with both Z values of -3.36, P<0.05). The protein expressions of keratin 10 in wound tissue of rats in control group and diabetes+metformin group on POD 7 as well as that in diabetes+metformin group on POD 13 were significantly lower than those in diabetes group ( P<0.05), and the protein expressions of PCNA in wound tissue of rats in control group and diabetes+metformin group on POD 7 were significantly higher than that in diabetes group ( P<0.05). There was a significant positive correlation between the relative expression of keratin 10 in wound tissue and the wound healing rate in control group and diabetes+metformin group of rats (with r values of 0.78 and 0.71, respectively, P<0.05), there was a significant negative correlation between the relative expression of PCNA in wound tissue and the wound healing rate in diabetes+metformin group of rats ( r=-0.60, P<0.05), and there was a significant negative correlation between the relative expressions of PCNA and keratin 10 in wound tissue of rats in diabetes group and diabetes+metformin group (with r values of -0.41 and -0.49, respectively, P<0.05). Conclusions:The diabetic rats with full-thickness skin defect wound exhibit delayed healing, accompanied by up-regulation of keratin 10 and down-regulation of PCNA in keratinocytes in the wound tissue. Metformin can promote wound healing in diabetic rats with full-thickness skin defects by down-regulating keratin 10 expression and up-regulating PCNA expression in keratinocytes in the wound tissue, and the wound healing rate was positively correlated with the expression of keratin 10 and negatively correlated with the expression of PCNA.