Objective To investigate the effect of normal saline(NS)of different temperature on morphological changes and nitric oxide synthase(NOS)expression of spinal cord.Methods The spinal canal of 96 SD adult rats was opened at T9.which of 24 rats was flushed with 37 ℃ NS,24 with 20 ℃ NS,24 with 4 ℃ NS respectively,and which of 24 rats in control group received no flushing.The spinal canal was closed one hour later,and the spinal cord was taken out 24 hours later.Then the water content in spinal cord was determined by dry-wet method.The morphological changes of spinal cord were observed under light microscope and the electronic microscope.The amount of NOS-positive neuron was measured by ?-NADPH histochemical methods.Results The water content in spinal cord was(66.53?0.61)% in control group,(66.75?1.00)% in 37 ℃ group,(70.55?0.77)% in 20 ℃ group,(71.92?2.50)% in 4 ℃ group.The spinal cord of control group and 37 ℃ group contained less water than that of 20 ℃ group and 4 ℃ group.There were no obvious morphological changes in the control group and 37 ℃ group.In 20 ℃ and 4 ℃ groups,the demyelination of axon,swelling of cell body and the disappearance of tigroid body were observed under light microscope,the partial disaggregation of medullary sheath,swelling of mitochondria and disappearance of mitochondria crista could be observed under electron microscope.The amount of NOS-positive neuron in spinal cord was(18.75?2.12),(18.63?1.41),(14.75?1.67),(8.13?1.25)in control,37 ℃,20 ℃ and 4 ℃ groups,respectively.The control group and 37 ℃ group showed more NOS-positive neuron than those of 20 ℃ group and 4 ℃ group.Conclusion NS below 20 ℃ can injury spinal cord.It is suitable to choose 37 ℃ NS to flush brain and spinal card during operation.

OBJECTIVE: To explore the genetic basis for a Chinese patient with retinitis pigmentosa (RP). METHODS: Whole exome sequencing (WES) was carried out to screen potential variant in the proband. Candidate variants were determined by taking consideration of clinical phenotype. Sanger sequencing was used to verify the variant in the proband and his parents. RESULTS: The proband was found to harbor compound heterozygous variants of c.8G>A (p.Cys3Tyr) and c.958_959insA (p.Arg320Glnfs*29) in the C2ORF71 gene, which has derived from his father and mother, respectively. Both variants were unreported previously. Based on the ACMG guidelines, they were predicted to be likely pathogenic and pathogenic, respectively. CONCLUSION The novel compound heterozygous variants of the C2ORF71 gene probably underlay the pathogenesis of RP in the proband. Above finding has enriched the spectrum of C2ORF71 gene mutations and facilitated genetic counseling for the family.
Asians/genetics* ; China ; Humans ; Mutation ; Pedigree ; Retinitis Pigmentosa/genetics* ; Whole Exome Sequencing

OBJECTIVE: To analyze the clinical characteristics and genetic features of a family affected with isolated proteinuria. METHODS: Clinical data of the family was collected. Mutations of 191 renal disease-related genes in the proband were screened with next generation sequencing (NGS). Sanger sequencing was used to verify suspected mutations in his family members and 100 healthy controls. The impact of the mutation was predicted with online software SIFT. Frequency of the mutation was searched in databases including 1000 Genomic Project, ESP and ExAC. RESULTS: NGS and Sanger sequencing showed that the proband harbored compound heterozygous mutations of ADCK4 gene including c.748C>G (p.Asp250His) and c.1041G>T (p.Cys347*), which were respectively inherited from his mother and father whom were both non-symptomatic. CONCLUSION The proband may have ADCK4-associated glomerulopathy due to the compound heterozygous mutations of the ADCK4 gene.
DNA Mutational Analysis ; Family ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Proteinuria ; genetics

OBJECTIVETo identify protein-protein interaction partners of PER1 (period circadian protein homolog 1), key component of the molecular oscillation system of the circadian rhythm in tumors using bacterial two-hybrid system technique.

METHODSHuman cervical carcinoma cell Hela library was adopted. Recombinant bait plasmid pBT-PER1 and pTRG cDNA plasmid library were cotransformed into the two-hybrid system reporter strain cultured in a special selective medium. Target clones were screened. After isolating the positive clones, the target clones were sequenced and analyzed.

RESULTSFourteen protein coding genes were identified, 4 of which were found to contain whole coding regions of genes, which included optic atrophy 3 protein (OPA3) associated with mitochondrial dynamics and homo sapiens cutA divalent cation tolerance homolog of E. coli (CUTA) associated with copper metabolism. There were also cellular events related proteins and proteins which are involved in biochemical reaction and signal transduction-related proteins.

CONCLUSIONIdentification of potential interacting proteins with PER1 in tumors may provide us new insights into the functions of the circadian clock protein PER1 during tumorigenesis.

Base Sequence ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Neoplasms ; genetics ; metabolism ; Period Circadian Proteins ; genetics ; metabolism ; Protein Binding ; Proteins ; genetics ; metabolism ; Two-Hybrid System Techniques

Autoantibody-related congenital heart block (ACHB) is a passively acquired autoimmune disease developing in fetuses after exposuring to maternal anti-Ro/Sj?gren's syndrome type A (SSA) antibody and/or anti-La/SSB antibody transported across the placenta, which contributes to fetal heart conduction system damage and signal conduction block at the atrioventricular node. However, fetal atrioventricular block does not necessarily occur with the presence of maternal autoantibodies, indicating its complex pathogenesis. This review focuses on the theories of calcium channels and apoptosis, the influence of other maternal factors and environmental changes on ACHB and the roles of natural killer cells and human leukocyte antigen in ACHB, aiming to provide reference for further study on the pathogenesis.

Objective:To evaluate the cardiac function and systolic dyssynchrony of fetuses exposed to maternal autoimmune antibodies (anti-SSA/Ro60, anti-SSA/Ro52 and anti-SSB/La) by using two-dimensional speckle tracking imaging (2D-STI).Methods:A total of 52 pregnant women with singleton pregnancy in the Affiliated Hospital of Inner Mongolia Medical University from July 2018 to November 2020 were selected. Eighteen fetuses of mothers with autoimmune antibodies were enrolled as autoimmune disease (AD) group and 34 fetuses of healthy mothers without antibodies were included as control group. Maternal baseline characteristics, fetoplacental Doppler parameters, and conventional echocardiographic data of two groups were prospectively collected. The systolic global and regional longitudinal strain of left and right ventricles (LV and RV) and the time to peak strain of regional myocardium were measured using 2D-STI. The differences in time to peak strain between the LV free wall and RV free wall (two-chamber dyssynchrony, 2C-DYS) and between the septum and LV free wall (one-chamber dyssynchrony, 1C-DYS) were also calculated.Results:There were no significant differences between the two groups in conventional systolic and diastolic functional parameters for the LV and RV(all P>0.05). The myocardial deformation parameters and 2C-DYS obtained by 2D-STI showed no statistical differences between two groups(all P>0.05). However, 1C-DYS was significantly more prolonged in the AD group than control group[28.50(13.50, 39.25)ms vs 19.50(8.00, 29.25)ms, P=0.042]. Conclusions:LV systolic mechanical dyssynchrony in fetuses of mothers with autoimmune antibodies suggests in-utero subclinical damage of the cardiac conduction system.

Objective:To observe the feasibility of evaluating right ventricular (RV) function by the method based on left ventricle (LV) pressure-strain ring (PSL) technique in patients with systemic lupus erythematosus (SLE), and to evaluate the efficacy of non-invasive PSL in the diagnosis of dysfunction of both LV and RV.Methods:Thirty-nine patients were enrolled with SLE who were admitted to the Affiliated Hospital of Inner Mongolia Medical University from March 2020 to September 2021 and 57 sex- and age-matched healthy controls underwent assessment by conventional echocardiography, two-dimensional speckle tracking imaging (2D-STI) and myocardial work (MW). The echocardiographic parameters were compared between the groups. The correlations between RVMW parameters and conventional echocardiographic parameters, and between MW parameters and SLEDAI-2K score were analyzed by Spearman correlation analysis.Results:①The two groups did not show any difference at the conventional echocardiographic parameters of the LV systolic function ( P>0.05). In the SLE group, tricuspid regurgitation maximal velocity (TR V max) and right ventricular index of myocardial performance (RIMP) increased, tricuspid annular plane systolic excursion (TAPSE) and tricuspid annular S′ (S′ tv) decreased (all P<0.05), and others did not show statistical difference (all P>0.05). ②The 2D-STI parameters: There were no significant differences in left ventricular global longitudinal strain (LV GLS) ( P>0.05), however RVGLS decreased ( P<0.05). ③The MW parameters of LV and RV: including global constructive work (GCW), global work index (GWI), global wasted work (GWW), and global work efficiency (GWE). In SLE group, LV GWE, RV GWE decreased and LV GWW, RV GWW increased ( P<0.05), others were no significant differences ( P>0.05). ④The correlation analysis: RV GWI positively correlated with TR V max ( rs=0.481). RV GCW positively correlated with TR V max ( rs=0.587). RV GWW positively correlated with right atrial area (RAA) and TR V max ( rs=0.429, 0.610), negatively correlated with S′ tv ( rs=-0.342). RV GWE positively correlated with S′ tv ( rs=0.326) and negatively correlated with RAA ( rs=-0.462) (all P<0.05). SLEDAI-2K score was negatively correlated with LV GLS ( rs=-0.333) and LV GWI ( rs=-0.326) ( P<0.05). Conclusions:The study has shown the impaired systolic function of both LV and RV in patients with SLE. PSL technique, especially RVMV based on LV PSL technique might emerged as a novel measure to evaluate systolic dysfunction in SLE patients.

The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)‍-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.
CRISPR-Associated Protein 9/metabolism* ; CRISPR-Cas Systems ; DNA ; Gene Editing/methods* ; Humans ; Streptococcus pyogenes/metabolism*