Objective:To diagnose and explore the genetic etiology of a neonate with Hereditary epidermolysis bullosa.Methods:A neonate who was admitted to Suqian Hospital Affiliated to Xuzhou Medical University on July 10, 2021 was selected as the study subject. Peripheral blood samples were collected from the child and his parents for the extraction of genomic DNA. And target gene capture and next-generation sequencing were carried out. Candidate variants were verified by Sanger sequencing and pathogenicity analysis.Results:The child was found to harbor compound heterozygous variants of the COL17A1 gene, namely c. 997C>T (p.Q333X) and c. 3481dupT (p.Y1161fs*2), which were respectively inherited from his father and mother. Both variants were predicted to be pathogenic. Conclusion:The child was diagnosed with Generalized atrophic benign epidermolysis bullosa due to the compound heterozygous variants of the COL17A1 gene.

Blood brain barrier (BBB) injury is the main pathological manifestation of many neurological diseases. Glycocalyx is the gel layer covering the lumen side of vascular endothelial cells, which plays an important role in regulating BBB function. However, glycocalyx is very fragile and easily damaged in various neurological diseases, leading to BBB destruction. This article focuses on the potential role of glycocalyx in cerebrovascular disease, the possible mechanisms related to glycocalyx and BBB injury, and explores the potential therapeutic strategies for protecting and restoring endothelial glycocalyx.

Objective:To investigate T cell epitope spectrum of alpha fetoprotein (AFP) restricted by 13 dominant human leukocyte antigen A (HLA-A) molecules in Chinese population.Methods:AFP T cell epitope candidates presented by 13 HLA-A molecules were in silico predicted using multiple epitope prediction algorithms. Then, the candidate epitope peptides were co-cultured in vitro with fresh peripheral blood mononuclear cells (PBMCs) from hepatocellular carcinoma (HCC) patients, followed by intracellular cytokine staining (ICS) and flow cytometry to verify the immunogenicity of the candidate epitope peptides. Peptide competition binding assay of HLA-A molecules were performed using 12 HMy2.CIR cell lines expressing the indicated HLA-A molecules to analyze the affinity and cross-restriction of candidate epitope peptides with HLA-A molecules. Results:PBMCs from 67 AFP + HCC patients were co-cultured in vitro with 42 candidate T cell epitope peptides, and the result showed that 20 epitope peptides stimulated CD8 + T cell responses (named as positive epitope peptides). Peptide competitive binding assay revealed 22 candidate epitope peptides with high affinity, 13 with inter affinity, six with low affinity, and 10 without affinity with indicated HLA-A molecules, respectively. The 20 positive epitope peptides presented high or inter affinity with corresponding HLA-A molecules and most of them displayed cross-binding properties with several HLA-A molecules. Conclusions:Twenty AFP-specific CD8 + T cell epitope peptides restricted by the predominant HLA-A molecules in Chinese population are obtained by the HCC patients-derived T-cell functional experiments, and the cross-binding ability of these epitope peptides to the corresponding HLA-A molecules is preliminary identified. These results provide systematic and fundamental data for the design of AFP-specific T cell detection systems and T cell epitope-based vaccines universal for the Chinese population.

OBJECTIVE: To explore the genetic basis for a neonate affected with Glutaric aciduria type I (GA-I). METHODS: Targeted capture and high-throughput sequencing was carried out for the proband and her parents. Candidate variants were verified by Sanger sequencing. RESULTS: The proband was found to harbor compound heterozygous variants of the GCDH gene, namely c.523G>A and c.1190T>C, which was derived from her father and mother, respectively. CONCLUSION The compound heterozygous variants of the GCDH gene probably underlay the GA-I in the patient.
Amino Acid Metabolism, Inborn Errors/genetics* ; Brain Diseases, Metabolic/genetics* ; Child ; Female ; Glutaryl-CoA Dehydrogenase/genetics* ; High-Throughput Nucleotide Sequencing ; Humans ; Infant, Newborn ; Mutation

O'Sullivan-Mcleod syndrome is a very rare variant of MND with a good prognosis. Its clinical feature is distal lower motor neuron syndrome of both upper limbs, and there is no effective treatment at present. We reported a case of O'Sullivan-Mcleod syndrome in this paper.The patient exhibited with middle-aged progressive distal muscle weakness and atrophy of both upper limbs, without sensory, cognitive or behavioral impairment and without pyramidal tract sign. Laboratory examination, imaging and genetic tests showed no obvious abnormalities. EMG revealed neurogenic damage to the small muscles of both hands. Now we retrospectively analyzed the clinical features of a patient with O'Sullivan-McLeod syndrome, and data from 18 cases for comparative analysis, in order to improve its understanding by clinicians.

Objective To investigate the expression of S100A16 in pancreatic cancer and its clinical significance. Methods Immunohistochemical experiment was used to detect the expression of S100A16 protein in pancreatic cancer tissues and adjacent tissues, and we analyzed the relation between S100A16 positive expression and clinicopathological parameters, prognosis of pancreatic cancer patients. PPI was used to predict a protein relationship network that directly interacted with S100A16. Results The positive rate of S100A16 expression in cancer tissues was significantly higher than that in adjacent tissues (P=0.001). S100A16 expression was higher in patients with vascular infiltration and lymph node metastasis. The overall survival time of patients with pancreatic cancer was related to tumor size, TNM stage and S100A16 expression level. Multivariate analysis showed that the expression level of S100A16 was an independent risk factor for the prognosis of pancreatic cancer patients, and the risk of death in patients with high S100A16 expression increased by 1.5 times. PPI predicted that S100A16 and S100A14, IL36G, PITX1, PERP played an important role in the interaction network. Conclusion Pancreatic cancer patients with high S100A16 expression have poor prognosis. The positive expression of S100A16 is an independent prognostic indicator.

Objective:To investigate the effect of virtual-simulation laparoscopic training system in the training of basic skills of endoscopic surgery in trainees with different clinical experience.Method:s Eight refresher physicians, eight residents who received standardized residency training, and eight undergraduate interns in five-year clinical medicine were selected. All of them received the training of endoscopic operations with the laparoscopic training box and the virtual-simulation laparoscopic training system for 30 minutes every day for 4 weeks. Data analyses were performed using SPSS 19.0 and t-test was adopted to compare the scores before and after training among the three groups.Result:s Before training, there were significant differences in endoscopic operations between the undergraduate intern group and the other two groups ( P<0.05); after 4 weeks of training, all three groups had significant increases in the scores and spent less time on training items ( P<0.05). There was no significant difference in simple operations among the three groups ( P>0.05), and the undergraduate intern group had a significantly higher score of complex operations than did the standardized residency training group and the refresher physician group ( P<0.05). Conclusion:The virtual-simulation laparoscopic training system can improve the laparoscopic skills of clinical trainees at different levels, therefore, it is worth being promoted in the teaching of clinical skills.

OBJECTIVE: To investigate the effects of etomidate on electrophysiological properties and nicotinic acetylcholine receptors (nAChRs) of ventral horn neurons in the spinal cord. METHODS: The spinal cord containing lumbosacral enlargement was isolated from 19 neonatal SD rats aged 7-12 days. The spinal cord were sliced and digested with papain (0.18 g/30 mL artificial cerebrospinal fluid) and incubated for 40 min. At the ventral horn, acute mechanical separation of neurons was performed with fire-polished Pasteur pipettes, and perforated patch-clamp recordings combined with pharmacological methods were employed on the adherent healthy neurons. In current-clamp mode, the spontaneous action potential (AP) of the ventral horn neurons in the spinal cord was recorded. The effects of pretreatment with different concentrations of etomidate on AP recorded in the ventral horn neurons were examined. In the voltage-clamp mode, nicotine was applied to induce inward currents in the ventral horn neurons, and the effect of pretreatment with etomidate on the inward currents induced by nicotine were examined with different etomidate concentrations, different holding potentials and different use time. RESULTS: The isolated ventral horn neurons were in good condition with large diverse somata and intact processes. The isolated spinal ventral horn neurons (=21) had spontaneous action potentials, and were continuously perfused for 2 min with 0.3, 3.0 and 30.0 μmol/L etomidate. Compared with those before administration, the AP amplitude, spike potential amplitude and overshoot were concentration-dependently suppressed ( < 0.01), and spontaneous discharge frequency was obviously reduced ( < 0.01, =12). The APs of the other 9 neurons were completely abolished by etomidate at 3.0 or 30 μmol/L. At the same holding potential (VH=-70 mV), pretreatment with 0.3, 3.0 or 30.0 μmol/L etomidate for 2 min concentration-dependently suppressed the current amplitude induced by 0.4 mmol/L nicotine ( < 0.01, =7). At the holding potentials of - 30, - 50, and - 70 mV, pretreatment with 30.0 μmol/L etomidate for 2 min voltage-dependently suppressed the current amplitude induced by 0.4 mmol/L nicotine ( < 0.01, =6 for each holding potential). During the 6 min of 30.0 μmol/L etomidate pretreatment, the clamped cells were exposed to 0.4 mmol/L nicotine for 4 times at 0, 2, 4, and 6 min (each exposure time was 2 s), and the nicotinic current amplitude decreased gradually as the number of exposures increased. But at the same concentration, two nicotine exposures (one at the beginning and the other at the end of the 6 min pretreatment) resulted in a significantly lower inhibition rate compared with 4 nicotine exposures ( < 0.01, =6). CONCLUSIONS etomidate reduces the excitability of the spinal ventral neurons in a concentration-dependent manner and suppresses the function of nAChR in a concentration-, voltage-, and use-dependent manner.
Animals ; Animals, Newborn ; Etomidate ; Neurons ; Patch-Clamp Techniques ; Rats ; Spinal Cord

Objective To investigate the protective effect of glibenclamide on neurovascular units (NVUs) and its possible mechanism in cerebral ischemia/reperfusion injury models.Methods One hundred and twenty healthy male SD rats were randomly divided into sham-operated group, model group, and glibenclamide (GBC) treatment group (n=40). Two h reperfusion models of acute focal middle cerebral artery occlusion were prepared by thread occlusion in rats of the latter two groups; rats in the model group were treated with 0.05％ DMSO saline solution two h after ischemia, and rats in the GBC treatment group were given intraperitoneal injection of 10μg/kg GBC with single dose. Immunofluorescence and Western blotting were used to detect the protein levels of sulfonylurea receptor 1 (SUR1) and transient receptor potential cation channel subfamily M member 4 (TRPM4) 8 h after reperfusion, and ELISA was used to detect the plasma level of matrix metalloproteinase 9 (MMP-9). At 24 h after reperfusion, Zea Longa scale was used to determine the neurological deficits; water content in the brain tissues was detected by dry and wet weight method, and blood-brain barrier (BBB) permeability was detected by Evans blue (EB) staining; Nissl staining was employed to detect the survival neurons; ionized calcium bindingadaptor molecule-1 (Iba-1) and cyclooxygenase-2 (COX-2) positive cells and IgG seepage quantity were detected by immumohistochemical staining to assess the neuro-vascular inflammation; the expressions of heat shock protein 70 (HSP70), phosphorylated protein kinase B (p-Akt), phosphorylated c-jun amino-terminal kinase (p-JNK), and phosphatidylinositol-3 kinase (PI3K) were detected by Western blotting.Results (1) At 8 h after reperfusion, the protein expressions of SUR1 and TRPM4 in the brain tissues of the model group were significantly increased as compared with those of the sham-operated group (P<0.05), and the two proteins were co-located; as compared with those in the model group, the protein expressions of SUR1 and TRPM4 in GBC treatment group was decreased, but the differences were not statistically significant (P>0.05). As compared with the sham-operated group, the model group had significantly higher MMP-9 level (P<0.05); as compared with the model group, the GBC treatment group had significantly lower MMP-9 level (P<0.05). (2) At 24 h after reperfusion, as compared with the sham-operated group, the model group had significantly increased Zea Longa scale scores, statistically increased brain water content, significantly increased EB permeability, significantly increased IgG seepage quantity, significantly smaller number of Nissl's staining-positive neurons, significantly larger number of Iba-1, COX-2 positive cells, and significantly decreased protein expressions of HSP70 and p-Akt (P<0.05); as compared with the model group, the GBC treatment group had significantly decreased Zea Longa scores, statistically decreased brain water content, significantly decreased EB permeability, significantly decreased IgG seepage quantity, significantly larger number of Nissl's staining-positive neurons, significantly smaller number of Iba-1, COX-2 positive cells, and significantly increased protein expressions of HSP70 and p-Akt (P<0.05).Conclusion SUR1-TRPM4 expression is increased after cerebral ischemia/reperfusion injury, and inhibition of SUR1-TRPM4 with GBC shows a protective role in NVUs after cerebral ischemia/reperfusion injury, possibly by regulating HSP70/p-Akt/MMP-9/COX-2 inflammatory signal pathway.

Objective To evaluate the changes ofcerebrovascular reserve (CVR) of patients with obstructive sleep apnea hypopnea syndrome (OSAHS) by transcranial cerebral doppler (TCD) and to study its relation with stroke.Methods One hundred and twenty-six patients with OSAHS,admitted to our hospitals from July 2012 to January 2013 and diagnosed as having OSAHS,were chosen in our study;they were divided into mild OSAHS group (n=49),moderate OSAHS group (n=44) and severe OSAHS group (n=33) according to test results of polysomnography (PSG).Another 40 healthy controls were collected.The CVR of all subjects were evaluated by TCD merging with CO2 experiment and compared among different groups.All the subjects were accepted continued two-year follow-up and recorded the accidents of stroke.The morbidities of stroke were compared between different groups.Results As compared with those in the control group and mild OSAHS group,all of the contractile reserve,dilatation reserve and overall reserve descended in moderate OSAHS group and severe OSAHS group with statistically significant differences (P＜0.05).As compared with those in the moderate OSAHS group,all of the contractile reserve,dilatation reserve and overall reserve descended in severe OSAHS group,and the differences were statistically significant (P＜0.05).In the two years of follow-up,the incidence of stroke in the severe OSAHS group (12.12％) was significantly higher than that in the control group (0％),mild OSAHS group (0％) and moderate OSAHS group (4.55％,P＜0.05).Conclusion The heavier the condition of OSAHS,the more obviously descended the CVR;the stroke morbidity of severe OSAHS patients is increased significantly.