Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Humans ; Apolipoprotein E4/genetics* ; Cytosine ; Mutation ; Blastocyst ; Heterozygote ; Gene Editing ; CRISPR-Cas Systems

CRISPR-Associated Protein 9 ; Gene Editing ; CRISPR-Cas Systems/genetics*

Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.

Objective:To detect the characteristics of bacteria in the feces of patients with rheumatoid arthritis (RA) and to further discover the relationship between intestinal flora and the status of peripheral cytokine, which might be able to provide new ideas for clinical treatment.Methods:The bacterial diversity and abundance of 111 RA patients and 100 age-and gender-matched healthy controls (HC) were detected by 16S high-throughput sequencing platform and compared. Based on the 16S rDNA high-throughput sequencing platform, the 16S rDNA V3 region in the participants' fecal specimens were analyzed and compared to screen for different bacterial groups. Alpha diversity was analyzed by the mothur software and the screening for different flora was tested by using Mann-Whitney, and the relationship between intestinal flora and peripheral cytokines were analyzed, too.Results:There was no significant difference in gender ( χ2=0.005, P=0.947) and age ( t=0.728, P=0.467) between the two groups. Patients with RA had a lower chao1 index ( Z=-2.188, P=0.029) and ACE index ( Z=-2.078, P=0.038) of species richness, and the Shannon index ( Z=-2.064, P=0.039) and Simpion index ( Z=-2.064, P=0.039) of diversity index in the feces compared with those of HC. At the genus level, the relative abundance of Bifidobacterium ( Z=-2.388, P=0.017), Lactobacillus ( Z=-2.543, P=0.011), Clostridium sensu stricto ( Z=-3.842, P<0.01), Blautia ( Z=-2.064, P=0.039) , Clostridium Ⅺ ( Z=-2.682, P<0.01), Turicibacter ( Z=-2.437, P=0.015), Phascolarctobacterium ( Z=-3.524, P<0.01), Megasphaera ( Z=-2.87, P<0.01), Veillonella ( Z=-2.472, P=0.013), Citrobacter ( Z=-3.263, P<0.01) and Escherichia/Shigella ( Z=-4.265, P<0.01) in RA were significantly higher than those of HC ( P<0.05), Butyricimonas ( Z=-3.071, P=0.002), Odorbacter ( Z=-2.257, P=0.024), Blautia ( Z=-2.064, P=0.039), Clostridium_ⅩⅣb ( Z=-2.901, P<0.01), Lachnospiracea_incertae sedis ( Z=-2.159, P=0.031), Acetivibrio ( Z=-2.995, P<0.01), Butyricicoccus ( Z=-2.162, P=0.031) and Gemmiger ( Z=-2.949, P<0.01) relative abundance were significantly decreased in RA patients ( P<0.05). LEfSe analysis showed γ-proteobacteria and Lachnospiraceaehad the most significant difference between the two groups. Further, patients with high inflammatory cytokines such as IL-17 and TNF-α hada higher relative abundance of Prevotella. Conclusion:The diversity and abundance of intestinal flora in RA patients are significantly different from those of healthy population, which is closely related to the levels of inflammatory cytokines, suggesting imbalance of intestinal flora might be involved in the occurrence and development of RA.

Background and purpose:As the development of the completion of the human genome project (HGP), the research focus is turning to the gene function research. At present, the domestic experimental research on the apoptosis of K562 cells induced by antisense olignonucleotides is rare. This study was aimed to investigate the effect of human chronic myelogenou leukemia (CML) bcr-abl fusion gene antisense oligonucletides on autophagy and apoptosis of CMLK562 cells in vitro. Methods:By liposome as the carrier, K562 cells were transfected with the bcr-abl gene antisense olignonucleotides. Hoechst staining method was used to observe the apoptosis inducing effect of different concentrations of oligonucleotides, the expressions of LC3-Ⅱ, autophagy-related protein, were determined by the Western blot method, the cell cycles were determined by lfow cytometry (FCM), and JEM-4000EX electron microscope technology was used to detect the apoptosis morphological changes. The apoptosis was detected by DNA agarose gel electrophoresis. Results:Hoechst staining results showed that the bcr-abl gene antisense oligonucletides signiifcantly promoted the apoptosis of K562 cells in a certain concentration dependent manner. Western blot showed that the expression level of LC3-Ⅱwas obviously higher in bcr-abl gene antisense oligonucletides transfected group than the control group, showing a promoting effect on cell autophagy. FCM test results showed that bcr-abl gene antisense oligonucleotides transfected K562 cells showed obvious cell cycle arrest, visible obvious apoptosis morphology under the electron microscope, and DNA Ladder showed obvious apoptosis fragments. Conclusion:The bcr-abl gene antisense olignonucleotides can signiifcantly induce the cell apoptosis of K562. This study provides a new method for CML therapy.

Objective:To screen and characterize aptamers against BCR-ABL fusion protein.Methods:A 90bp single stranded DNA( ssDNA) random library was subjected to 13 rounds of selection against BCR-ABL fusion protein by systematic evolution of ligands by expotential enrichment ( SELEX ) method, the selected aptamers were cloned and sequenced.The primary sequences and structure of aptamers were analyzed by Clustal W and DNA Folding Sever and the percentage of the ssDNA pool bound to BCR-ABL core protein were determinated.Results: after 13 rounds selection, the percentage of ssDNA pool bound to BCR-ABL fusion protein increased from 0.3%to 47.1%,the results showed that affinities of the Aptamers were different,the second structure analysis revealed possible stem-loops for binding to BCR-ABL fusion protein,the affinity of aptamer A2 to BCR-ABL fusion protein was highest with Kd values as low as 72 nmol/L.Conclusion:Aptamers against BCR-ABL fusion protein has been identified by SELEX methods from a 90 bp single stranded DNA library.And provide certain reference for the clinical treatment of chronic myelogenous.

Objective:To retrospectively analyze the survival outcomes of the surgical management of primary small cell carcino-ma of the esophagus. Methods:The medical records were reviewed for patients diagnosed with esophageal carcinoma and underwent esophagectomy from January 2000 to December 2009 at the Department of Thoracic Surgery of the Henan Cancer Hospital. We fo-cused on the clinical data of patients with small cell carcinoma of the esophagus. The Kaplan-Meier approach with log-rank test was used for survival analysis. Results:A total of 5,062 patients underwent esophagectomy with curative intent at the Department of Thorac-ic Surgery of the Henan Cancer Hospital;among which, 57 (1.1%) were diagnosed with small cell carcinoma of esophagus. The most common surgical approach was trans-left thoracic incision esophagectomy. Cervical esophagogastrostomy was performed for all pa-tients. The most common chemotherapy regimen was EP. The overall 5-year survival rate was 12.5%, and the median survival time was 45 months. Among the various stages, the 5-year survival rate and survival time were 25% and 50 months for Stage I, 5.9% and 43 months for Stage II, and 4.3%and 43 months for StageⅢ. Subgroup analysis showed that cases treated with surgery alone had poorer overall median survival time compared with those cases that underwent surgery plus chemotherapy (23.2 months vs. 60.7 months, re-spectively;P<0.01). Even for Stage I patients, thesurgery plus chemotherapysubgroup was associated with a significantly longer me-dian survival time than the surgery alone subgroup (81.9 months vs. 22.3 months, P<0.01). Conclusion:For patients with primary small cell carcinoma of the esophagus, surgery alone cannot provide the optimal prognosis. Surgery combined with systemic chemother-apy can improve the survival time.

Hexanes ; adverse effects ; Humans ; Occupational Injuries ; chemically induced ; Pyrroles ; adverse effects ; Trauma, Nervous System ; chemically induced

BACKGROUND:The monocyte chemoattactant protein-1 gene polymorphism is associated with spinal tuberculosis susceptibility. OBJECTIVE:To investigate the association between serum monocyte chemoattactant protein-1 expression level and spinal tuberculosis susceptibility in Han population of Hunan province. METHODS:The patients with spinal tuberculosis and the healthy volunteers were recruited in Xiangya Hospital of Central South University from December 2004 to December 2010. The empty peripheral venous blood 2 mL were col ected from the subjects in early morning, then the monocyte chemoattactant protein-1-362 genotypes were detected by polymerase chain reaction and DNA sequencing technology. And the serum monocyte chemoattactant protein-1 level was detected by enzyme linked immunosorbent assay technology. The ROC curve was used for diagnostic tests to calculate diagnostic threshold value of serum monocyte chemoattactant protein-1 level to spinal tuberculosis susceptibility, and to analyze the diagnostic titer. RESULTS AND CONCLUSION:208 patients with spinal tuberculosis and 210 healthy volunteers were included. The serum monocyte chemoattactant protein-1 level of the spinal tuberculosis patients was significantly higher than that of the healthy volunteers [(134.58±51.63) ng/L vs. (39.18±17.45) ng/L, P<0.01]. The serum monocyte chemoattactant protein-1 level could not be affected by gender, but over-expressed in patients with monocyte chemoattactant protein-1-362-CC genotypes. The serum monocyte chemoattactant protein-1 level higher than 101.65 ng/L indicated that the patients might suffered from spinal tuberculosis (sensitivity:85.5%, specificity:94.3%, Youden index:0.799, area under curve of ROC:0.946, 95%confidence interval:0.916-0.975, P<0.01). The serum monocyte chemoattactant protein-1 level may be associated with spinal tuberculosis susceptibility in Han population of Hunan province, highly expressed serum monocyte chemoattactant protein-1 can be used as one of the indicators for the diagnosis of spinal tuberculosis.

This study examined the wave intensity (WI) of the carotid artery in patients with hyperthyroid in order to assess the hemodynamic changes of hyperthyroid patients. A total of 86 hyperthyroid patients without cardiac morphological changes and arrhythmia, and 80 healthy control subjects were enrolled in the study. Right common carotid artery (RCCA) was selected for ultrasonic imaging to obtain WI indices, including amplitude of the peak during early systole (W1), amplitude of the peak during late systole (W2), area of the negative wave during mid-systole (NA), interval between R wave of electrocardiogram and W1 (R-1st), interval between W1 and W2 (1st-2nd). The levels of serum thyroid hormones, consisting of free triiodothyronine (FT3), free thyroxin (FT4) and thyroid stimulating hormone (TSH), were measured in hyperthyroid patients. Echocardiographic indices including left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were determined in each subject. The results showed that the W1, W2, NA, and (1st-2nd×HR) in hyperthyroid patients were significantly higher than those in healthy controls. There was no significant difference in LVEF and LVFS between the two groups. FT3 was correlated with W1, W2, NA, (1st-2nd×HR), pulse pressure (PP) and heart rate (HR) in hyperthyroid patients. Several abnormal waves on WI curves were present in 19 hyperthyroid patients during mid-systole. It was concluded that WI technique may prove a real-time, noninvasive, sensitive and convenient tool for assessing the cardiac function and hemodynamic alterations in hyperthyroid patients.