Identification of pathogenic mutant genes in seven families with primary biliary cholangitis patients by whole exome sequencing / 中华消化杂志

Objective:To screen the common low-frequency mutation sites in primary biliary cholangitis (PBC) by whole exome sequencing (WES), in order to find PBC-related new susceptibility genes.Methods:From January 2000 to December 2017, the clinical data of seven patients with PBC of three PBC families diagnosed at General Hospital of Tianjin Medical University and two healthy controls were collected. The DNA blood samples were extracted and analyzed by WES. SAMtools 1.3 software was used to detect gene single nucleotide polymorphism (SNP) and indel sites, and gene mutation sites were screened from known databases of 1000 Genome, ExAC, ESP6500 and Novo-Zhonghua gene database. Pymol V2.3.2 software was performed to simulate the three-dimensional structure of major histocompatibility complex-Ⅱ (MHC-Ⅱ), and the amino acid position corresponding to the common mutation sites among families were observed.Results:The age of first diagnosis of seven PBC patients was (61.2±10.2) years. The results of serum test of seven patients indicated that alkaline phosphatase (ALP) level was (306.9±242.5) U/L, γ-glutamyltranspeptidase (GGT) level was (121.7±85.9) U/L, alanine aminotransferase (ALT) level was (47.6±33.1) U/L, aspartate aminotransferase (AST) level was (55.7±34.1) U/L and immunoglobulin G level was (14.9±3.1) g/L. The antinuclear antibody were all cytoplasmic granule types and anti-mitochondrial antibody were all positive. Five PBC patients developed intra-abdominal lymphadenopathy; two patients had extrahepatic autoimmune diseases and the pathological results of liver biopsy of two patients both showed interface hepatitis and small bile duct lesions. Eighteen SNPs were common in three PBC families, which were located in the gene of OTOA, OBSCN and human leucocyte antigen- DRB1( HLA- DRB1). rs200988634 located in OTOA gene was a common polymorphic locus among the three families. rs746424683, rs545316651, rs553144914, rs533059830 and rs56087721 located in OBSCN caused the changes of nine amino acids of different location. There were 12 SNP variations located in HLA- DRB1 gene, which leaded to the changes of 12 amino acids of different location, among them rs16822698, rs112796209 and rs11554463 mutation induced G154A, Y152C and Y107X amino acid variation of MHC-Ⅱ beta chain, and Y107X amino acid was located in the groove region of MHC-Ⅱ binding with peptide. Conclusions:WES in PBC families is a good strategy to elucidate the candidate deleterious mutation genes OBSCN and OTOA. HLA- DRB1 which is a susceptible gene of PBC may affect MHC-Ⅱ mediated antigen presentation process by the changing amino acid sequence.

Clinical characteristics of patients with autoimmune liver disease complicated with gallbladder stone / 中华消化杂志

Objective:To explore the clinical characteristics of liver function of patients with autoimmune liver disease (AILD) complicated with gallbladder stone (GS), so as to guide clinical practice.Methods:From November 2009 to October 2018, at General Hospital of Tianjin Medical University, the clinical data of 386 patients with AILD were retrospectively analyzed. According to the relevant diagnostic criteria, 208 cases of autoimmune hepatitis (AIH), 129 cases of primary biliary cholangitis (PBC) and 49 cases of PBC-AIH overlap syndrome were screened out. The incidence, clinical characteristics and the changes of laboratory indicators including albumin, alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT) of AILD patients complicated with GS were analyzed. Chi-square test, t test and rank sum test were performed for statistical analysis. Results:There was no significant difference in the incidence between AILD, AIH, PBC and PBC-AIH overlap syndrome patients complicated with GS (32.9%, 127/386; 28.8%, 60/208; 36.4%, 47/129 and 40.8%, 20/49; respectively; P>0.05). Gallstones of AILD patients complicated with GS mostly were multiple and small stones with maximum diameter <1 cm (45.7%, 58/127 and 57.7%, 60/104, respectively). The age of initial diagnosis, the proportion of liver cirrhosis at inital diagnosis and the levels of ALP and GGT were higher in AILD patients complicated with GS than those of AILD patients without GS ((60.5±11.5) years vs. (57.6±11.5) years; 53.5%, 68/127 vs. 42.1%, 109/259; 154.00 U/L (89.00 U/L, 257.00 U/L) vs. 125.00 U/L (86.00 U/L, 212.00 U/L); 169.00 U/L (79.00 U/L, 343.00 U/L) vs. 128.60 U/L (48.00 U/L, 284.00 U/L); respectively); however the albumin level was lower than that of AILD patients without GS ((36.46±7.30) g/L vs. (38.34±7.58) g/L), and the differences were statistically significant ( t=-2.361, χ2=4.506, Z=-2.192, -2.443, t=2.322; all P<0.05). The incidence of GS in AILD patients≥60 years old was higher than that AILD patients<60 years old (37.6%, 73/194 vs. 28.1%, 54/192), and the difference was statistically significant ( χ2=3.948, P=0.047). The incidence of GS in AILD patients and AIH patients complicated with liver cirrhosis was higher than that in patients without liver cirrhosis (38.4%, 68/177 vs. 28.2%, 59/209; 35.7%, 35/98 vs. 22.7%, 25/110; respectively), and the differences were statistically significant ( χ2=4.506 and 4.259, P=0.034 and 0.039). Conclusions:AILD patients complicated with GS are common, most are multiple and small stones. When complicated with GS, the initial diagnosis may be delayed and the rate of liver cirrhosis at initial diagnosis may increase. The incidence of GS is high in AILD patients with older age and liver cirrhosis.

Effect of Motor Imagery on Motor Function in Hemiplegic Patients after Stroke / 中国康复理论与实践

Objective To explore the effect of motor imagery on motor recovery in hemiplegic patients after stroke. Methods From May, 2015 to October, 2016, 40 hemiplegic patients after stroke were randomly divided into control group (accepted routine rehabilitation, n=20) and motor imagery group (accepted motor imagery and routine rehabilitation, n=20). They were assessed with Fugl-Meyer Assess-ment (FMA), modified Barthel Index (MBI) before and six weeks after treatment. Fractional anisotropy (FA) of the focus was measured with diffusion tensor imaging (DTI). Results The scores of FMA and MBI improved in both groups after treatment (t>5.088, P2.124, P3.892, P0.05). FA increased in more patients of the motor imagery group (5/5) than in the control group (2/4). Conclusion Motor imag-ery can promote the recovery of motor function and activities of daily living in stroke patients, and may help the recovery of fibers in white matter.

Level of Serum Vascular Endothelial Growth Factor in Patients with Cerebral Infarction before and after Rehabilitation / 中国康复理论与实践

@#Objective To investigate the change of serum vascular endothelial growth factor (VEGF) in cerebral infarction before and af-ter rehabilitation. Methods Forty-eight patients with first cerebral infarction were enrolled from June, 2014 to August, 2016 in Beijing Bo'ai Hospital. Level of serum VEGF was measured by enzyme-linked immunosorbent assay (ELISA) before and six weeks after rehabilitation. The level of serum VEGF of 33 normal subjects was compared with the patients'. Results The level of serum VEGF was higher in the pa-tients group than in the control group before and six weeks after rehabilitation (t>2.540, P<0.05). The level of VEGF was higher in the large area infarction group (>4 cm) than in the small area infarction group (≤4 cm) (t=4.436, P<0.05), and was higher in the short course (less than one month) infarction group than in the long course (more than one month) group (t=2.316, P<0.05). Conclusion VEGF can be main-tained in high level after rehabilitation.

Rehabilitation Medicine Teaching for International Medical Students / 中国康复理论与实践

With the continuous expansion of the enrollment of international medical students and the improvement of teaching quality, as well as the rapid development of rehabilitation medicine in China, rehabilitation medicine has become one of the required courses for in-ternational medical students. In view of the main problems suffered by the international medical undergraduate students in the study of reha-bilitation medicine, we mainly focused on the education concepts, teaching management and quality evaluation system, curriculum setting, construction of teaching materials, teacher training, teaching mode, teaching research and so on. The aim is to provide reference for improv-ing education quality in the course of rehabilitation medicine for the international medical students.

Effects of Myelotomy on Autophagy in Injured Spinal Cord in Rats / 中国康复理论与实践

@#Objective To observe the effects of myelotomy on autophagy activation after traumatic spinal cord injury (SCI) in rats. Methods 54 adult female Sprague-Dawley rats were randomly assigned to sham-operated group (SG, n=18), contusion group (CG, n=18) or myelotomy group (MTG, n=18). The T10 SCI model in rats was induced with a New York University (NYU) impactor and myelotomy was performed 24 hours after SCI. They were evaluated with the BBB score 1, 7, 14 days after injury. The expression of mRNA of Beclin-1 and Bcl-2 were detected with real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The formation of autophagosome was investigated under electronic microscope (EM) 3 days after injury. Results BBB score was more in the MTG than in the CG 7 and 14 days after injury (P<0.05), while the expression of Beclin-1 mRNA was less (P<0.05). The expression of Bcl-2 mRNA was more in the MTG than in the CG 3 and 7 days after injury (P<0.05). The expression of Beclin-1 mRNA was negatively correlated with BBB scores (P< 0.05). The formation of autophagosome was less in the MTG than in the CG. Conclusion Myelotomy can improve the recovery of motor function in rats after acute traumatic SCI, which may associate with neuroprotection mediated by inhibition of autophagy through the Bcl-2 signaling pathway.

Effects of Myelotomy on Autophagy in Injured Spinal Cord in Rats / 中国康复理论与实践

Objective To observe the effects of myelotomy on autophagy activation after traumatic spinal cord injury (SCI) in rats. Methods 54 adult female Sprague-Dawley rats were randomly assigned to sham-operated group (SG, n=18), contusion group (CG, n=18) or myelotomy group (MTG, n=18). The T10 SCI model in rats was induced with a New York University (NYU) impactor and myelotomy was performed 24 hours after SCI. They were evaluated with the BBB score 1, 7, 14 days after injury. The expression of mRNA of Beclin-1 and Bcl-2 were detected with real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The formation of autophagosome was investigated under electronic microscope (EM) 3 days after injury. Results BBB score was more in the MTG than in the CG 7 and 14 days after injury (P<0.05), while the expression of Beclin-1 mRNA was less (P<0.05). The expression of Bcl-2 mRNA was more in the MTG than in the CG 3 and 7 days after injury (P<0.05). The expression of Beclin-1 mRNA was negatively correlated with BBB scores (P<0.05). The formation of autophagosome was less in the MTG than in the CG. Conclusion Myelotomy can improve the recovery of motor function in rats after acute traumatic SCI, which may associate with neuroprotection mediated by inhibition of autophagy through the Bcl-2 signaling pathway.

Myelotomy suppresses autophagic marker LC3-II expression and elevates mTORC1 expression and improves neurological function recovery in rats with gical function recovery in rats with

Although previous studies have shown functional efficacy of myelotomy for the treatment of spinal cord injury (SCI), the underlying mechanism remained unknown. This study aimed to determine the relationship between myelotomy-mediated neuroprotection and autophagy following SCI by evaluating the expression of microtubule-associated protein light chain 3 (LC3-II) and mammalian target of rapamycin complex 1 (mTORC1). Ninety-nine adult female rats were randomly assigned to either sham-operated group (SG), model group (MG), or 24 h-myelotomy group (MTG). SCI at T10 was induced with a New York University impactor, and myelotomy was performed 24 h after SCI. Functional recovery was evaluated via the open-field test. The protein expression of LC3-II was analyzed by Western blot, and the mRNA expression of LC3-II and mTORC1 were detected by real-time quantitative reverse transcriptase polymerase chain reaction. Rats in the MTG exhibited significantly better performance in the hind limbs compared to those in the MG on day seven and fourteen post-injury. Myelotomy suppressed the protein and mRNA expression of LC3-II on day three, seven and fourteen post-injury and increased the mRNA expression of mTORC1 in the MTG on day three and seven post-injury. The LC3-II protein expression was significantly and negatively correlated with BBB scores at day seven and fourteen post-injury. These results showed that myelotomy-induced neuroprotection in a rat model of SCI was likely mediated by inhibition of autophagy by activation of the mTORC1 signaling pathway