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

Biliary malignant tumors have an insidious onset and rapid development, and most patients have lost the opportunity for radical surgery at initial diagnosis and often have poor prognosis. Gemcitabine-based chemotherapy is the first-line treatment for biliary malignant tumors, but with a limited clinical effect. The improvement in next-generation sequencing technology provides the possibility for the precise treatment of biliary malignant tumors, but the application and development of the precise treatment of biliary malignant tumors are limited by the low positive rate of targets and the poor accessibility of therapeutic drugs. The advent of the era of immunotherapy represented by the immune checkpoint inhibitor PD1/PD-L1 monoclonal antibody brings a promising future for the treatment of malignant tumors, including biliary malignant tumors. Combined chemotherapy and/or targeted therapy based on immune checkpoint inhibitors has shown a good effect in the treatment of biliary malignant tumors, which is the direction of the treatment of advanced biliary malignant tumors in the future.

Objective:To correlate climate change with epistaxis in Yueqing of Zhejiang province and to provide evidence for the diagnosis and treatment of epistaxis.Methods:A total of 1 800 patients with epistaxis, who received treatment in Yueqing People's Hospital between October 2018 and October 2019, were included in this study. The data of these patients were input into the Microsoft Excel software. They were summarized and sorted as per admission time. Climate change was correlated with epistaxis.Results:Daily average temperatures were negatively correlated with epistaxis ( r = -0.65, P = 0.003). A non-linear, inverted-U-shaped relationship was observed between daily relative humidity and epistaxis. When the daily relative humidity was < 65%, daily relative humidity was positively correlated with epistaxis ( r = -0.54, P = 0.007).When the daily relative humidity was > 65%, daily relative humidity was negatively correlated with epistaxis ( r = -0.68, P = 0.002). There was a nearly linear positive correlation between diurnal temperature range and epistaxis ( r = 0.52, P = 0.009). There was a linear and positive correlation between daily average atmospheric pressure and epistaxis ( r = 0.60, P = 0.004). The risk of epistaxis increased by 1.48% (95% CI: -2.15 to -0.81) when the temperature decreased by 1 ℃. When daily relative humidity was < 65%, the effects of humidity change on the risk of epistaxis were not statistically significant ( P > 0.05). When the relative humidity of the day was > 65%, the risk of epistaxis decreased by 1.82% (95% CI: -2.71 to -0.93) for every unit of humidity rise. The risk of epistaxis increased by 2.86% (95% CI: 0.54 - 5.18) for every 1 ℃ increase in temperature. The risk of epistaxis increased by 1.18% (95% CI: 0.50 - 1.87) for every 1 Pa increase in air pressure. Conclusion:Temperature change is negatively correlated with epistaxis. Atmospheric pressure and diurnal temperature range are positively correlated with epistaxis. Temperature change, atmospheric pressure, and diurnal temperature have temporary effects on epistaxis. High humidity has an obvious long-term protective effect against epistaxis.

AIM: To study the inhibitory effect of Xihuang Pill on H

Objective:To investigate the predictive value of serum procalcitonin (PCT) for the localization of acute digestive tract perforation.Methods:This retrospective study included 88 patients from the Department of Gastrointestinal Surgery of the First Affiliated Hospital of Southwest Medical University who were diagnosed as acute digestive tract perforation between January 2015 and January 2018. According to the intraoperative diagnosis and postoperative pathological reports, the enrolled patients were divided into the upper digestive tract perforation group (45 cases) and the lower digestive tract perforation group (43 cases) (above or below Treitz ligament). Preoperative serum PCT, white blood cell, neutrophil rate, C-reactive protein (CRP), albumin (ALB), C-reactive protein to albumin (CRP/ALB) ratio were measured and compared between the two groups. Univariate analysis and multivariate logistic regression analysis were used to analyze the independent risk factors of the lower digestive tract perforation group, and the receiver operating characteristic curve was used to analyze the predictive value of the above mentioned markers in the localization of acute digestive tract perforation.Results:Univariate analysis showed that patients in the lower digestive tract perforation group exhibited significantly higher levels of serum PCT, CRP, ALB, and CRP/ALB ratio than patients in the upper digestive tract perforation group ( P<0.05). Multivariate logistic regression analysis showed that serum PCT, CRP and CRP/ALB ratio were independent risk factors for the diagnosis of lower digestive tract perforation [PCT: odds ratio ( OR)=1.241, 95% confidence interval (95% CI): 1.098~1.403, P = 0.001; CRP: OR= 0.95, 95% CI: 0.912~0.99, P = 0.014; and CRP/ALB ratio: OR= 35.104, 95% CI: 3.889-316.885, P = 0.002]. The area under curve of serum PCT, CRP, CRP/ALB ratio to distinguish upper or lower digestive tract perforation were 0.932 (95% CI: 0.879~0.985), 0.77 (95% CI: 0.667~0.872), and 0.898 (95% CI: 0.827~0.969), respectively. The optimal cutoff value of PCT in differential diagnosis of upper or lower digestive tract perforation was 16.595 ng/mL, with a sensitivity of 86.0% and a specificity of 91.1%. The optimal cutoff value of CRP was 55.4 mg/mL, with a sensitivity of 76.7% and a specificity of 80.0%. The optimal cutoff value of CRP/ALB ratio was 1.45 and its sensitivity and specificity were 83.7%, 88.9%, respectively. Conclusions:Serum PCT, CRP, and CRP/ALB ratio are helpful to predict the localization of acute digestive tract perforation and can improve the diagnostic accuracy. The diagnostic efficiency of PCT is better than CRP and CRP/ALB ratio, exerting excellent clinical value.

Objective To discuss the controversial role of breast milk in late-onset group B Streptococcus (GBS) infections.Methods This study reported a case of recurrent late-onset GBS sepsis with the suspicion of breast milk transmission in an extremely preterm infant born at 22+6 weeks who was treated at the University of Hong Kong-Shenzhen Hospital in September 2016.Literatures about late-onset GBS cases associated with contaminated breast milk were reviewed to investigate whether GBS could be transmitted through breast milk.Results (1) Case report:A breast-fed extremely preterm infant born at 22+6 gestational weeks suffered from GBS sepsis along with meningitis for the first time on 100 d.The mother was negative for rectovaginal GBS screening.Breast milk wasn't tested as no signs of mastitis were found.The neonate recovered from the first GBS sepsis after 14 days of antibiotic treatment,then returned to breastfeeding.On 126 d,GBS sepsis reoccurred in this baby.Fresh breast milk culture yielded GBS which was identical with the GBS strains isolated from the neonatal blood in antimicrobial susceptibility.After recovery from the second episode,the baby was partially breastfed again without further relapses of late-onset GBS sepsis.(2) Literature review:64 cases of late-onset GBS infections that transmitted via breast milk were retrieved from PubMed,while no Chinese cases had been reported.Clinical data of the 65 cases (including this case) were reviewed and the results revealed that contaminated breast milk was associated with late-onset GBS infections.The reported relapse rate of GBS infections transmitted via breast milk was 25％ for two episodes and 7％ for three episodes.Conclusions GBS contaminated breast milk could potentially cause late-onset GBS sepsis in infants and further studies are required to identify the underlying mechanisms.

Objective: To detect the expression of miR-133a-3p in gastric cancer (GC) tissues and plasma of GC patients, and to investigate its effect on the proliferation of GC cells as well as its correlation toprognosis of GC patients. Methods: 52 cases of cancertissues (non-necrosis part) and corresponding adjacent tissues as well as the pre-operative peripheral blood samples from GC patients, who underwent surgery at Department of General Surgery, the Forth Hospital of Hebei Medical University(Shijiazhuang, China) between May 2012 and May 2013, were collected for this study. The plasma sample (n=35) from healthy donors were obtained during their physical examination. RT-qPCR was adopted to detect the expression of miR-133a-3p in gastric cancer tissues, adjacent tissuesand plasma samples of GC patients and healthy volunteers. The relationships between miR-133a-3p expression and the median DFS as well as clinicopathological parameters were also analyzed. CCK-8 assay was adopted to detect the effect of miR-133a-3p silence or over-expression on proliferation of gastric cancer SGC7901 cells. Results: miR-133a-3p was dramatically decreased in gastric cancer tissues (P<0.01), and its expression was associated with TNM stage, tumor infiltration (T), lynphonode metastasis (N), and vascular tumor thrombus (all P<0.01); miR-133a-3p was significantly increased in the plasma of GC patients (P<0.01), and its expression was associated with TNM stage, lynphonode metastasis (N), and vascular tumor thrombus (all P<0.05). miR-133a-3p expression was positively correlated with serum CA199 level of GC patients (P<0.01). The median DFS of patients with high miR-133a-3pexpression in cancer tissues was significantly longer than that of the patients with low expression(20.8 vs 14.8 months, P<0.05); The median DFS of patients with high plasma miR-133a-3p expression was significantly shorter than that of the patients with low expression (14.4 vs 20.3 months, P<0.05). Over-expression of miR-133a-3p could significantly inhibit the proliferation of gastric cancer SGC7901 cells, while miR-133a-3p silence could significantly promote the proliferation (all P<0.05). Conclusion: miR-133a-3p could significantlyinhibit the proliferation of SGC7901 cells; miR-133a-3p aberrantlyexpressed in gastric cancer tissues and plasma, and obviously correlated with prognosis of gastric cancer patients, which may be used as a potential clinical bio-maker for early diagnosis and treatment as well as the prognosis prediction of gastric cancer.

Objective To explore the expression of methyl CpG binding protein 2 (MeCP2) in gastric cancer tissues and its role in multi-drug resistance (MDR) in gastric cancer cells.Methods The expression of MeCP2 in 90 gastric cancer tissues and the adjacent normal tissues was detected by immunohistochemistry,and the relationship between MeCP2 expression level and clinicopathological parameters was analyzed.The expression level of MeCP2 in gastric cancer cell line SGC7901,MDR cell variants SGC7901/doxorubicin hydrochloride(ADR) and SGC7901/vincristine(VCR)was determined by Western blot.After the expression of MeCP2 was silenced by short interference RNA (siRNA),half inhibitory concentration (IC50) of 5-fluorouraeil and cisplatin in gastric cancer cell was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT) assay.The t test or chi square test was performed for statistical analysis.Results The positive rate of MeCP2 expression in gastric cancer tissues was 72.2％ (65/90),which was significantly lower than that in adjacent normal tissues (93.3％,84/ 90),and the difference was statistically significant (x2 =14.068,P＜0.01).MeCP2 expression was not correlated with age,tumor maximum diameter and lymph node metastasis (all P＞0.05),however which was related to gender,clinical TNM stages and distant metastasis (x2=4.680,4.186 and 4.327;aH P＜ 0.05).The gray scale ratios of MeCP2/β-actin in SGC7901/ADR and SGC7901/VCR were 0.593 7 ± 0.030 5 and 0.651 2 ± 0.018 6,which were lower than that of parental SGC7901 cells (1),and the differences were statistically significant (t =23.080,17.360;both P ＜ 0.01).After the expression of MeCP2 was silenced by siRNA,the IC50 of 5-fluorouracil and cisplatin in SGC7901 transfected with MeCP2 specific siRNA were (11.540 0±0.469 3) μg/mL and (2.273 0±0.265 4) μg/mL,which were higher than the IC50 of 5-fluorouracil and cisplatin in SGC7901 transfected with non-related oligonucleotide sequence ((8.663 0±0.160 1) μg/mL and (0.884 0 ±0.038 6) μg/mL),respectively.The differences were statistically significant (t =15.380 and 8.153;both P ＜ 0.01).Conclusions The expression of MeCP2 in gastric cancer tissues significantly decreased,which was correlated with clinical stages,distant metastasis.MeCP2 can inhibit the MDR of gastric cancer cell,which indicated the dysregulated expression of MeCP2 might participate in the genesis and development of gastric cancer.

OBJECTIVETo investigate the relationship of the mutational status of the ND4 gene and the clinical features of acute myelogenous leukemia (AML) patients with ND4 mutations.

METHODSUsing PCR combined with directly sequencing, we identified somatic mutations of ND4 in 121 primary AML patients to couple with their clinical features.

RESULTSThere were 58 male patients and 63 female patients (median age 49 years, 10-86 years). Eight of 121 patients (6.6%) with de novo AML were found harboring missense mutation of ND4 gene, including 3 patients with A131V (3/8, 37.5%), 2 patients with A404T (2/8, 25%), 1 patient with F149L (1/8, 12.5%), 1 patient with G242D (1/8, 12.5%) and 1 patient with Y409H (1/8, 12.5%), respectively. Patients with ND4 mutations were associated with good karyotype (P=0.049), regardless of gender, age, white blood cell, hemoglobin, platelet, blast cells of bone marrow or immunophenotype (P>0.05). There were no statistical significance in mutations of FLT3-ITD, NPM1, CEBPA, c-KIT and DNMT3A between patients with ND4 mutation and wild-type (wt) ND4 (P>0.05). The median overall survival of patients with ND4 mutations and wt ND4 were all not reached. The median relapse-free survival were not reached and 29(2-53) months, respectively (P>0.05). There was no significance in the ratio of CR and RR patients between wt ND4 and ND4 mutated groups (P>0.05).

CONCLUSIONIt was concluded that novel ND4 mutations could be found in de novo AML patients, especially in patients with good karyotype. Thus, ND4 mutations might play an important role in AML prognosis. However, whether the mitochondria dysfunction contribute to leukemogenesis needs to be further investigated.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; Male ; Middle Aged ; Mutation ; NADH Dehydrogenase ; genetics ; Prognosis ; Young Adult

Objective To evaluate the effect of bronchoscopic argon plasma coagulation thera-py on bronchial carcinoma. Methods Thirty-one bronchial carcinoma patients were diagnosed by bronchoscope and pathological tests, with or without atelectasis or obstructive pneumonia on chest X-ray or chest CT. Argon plasma coagulation therapy was performed through bronchoscope. The location of the airway lesions, the degree of obstruction, dyspnea index, and complications were evaluated. Results The patients with bronchial carcinoma were treated 1～4 times by bronchoscopic argon plas-ma coagulation therapy. Full effectiveness was achieved in 15 patients (48.4 %), partial in 12 (38.7%), and mild in the other 4 ( 12.9 % ). The overall effective rate was 100 %. Conclu-sion Bronchoscopic argon plasma coagulation therapy for bronchial carcinoma can remarkably reduce the tumor size, relieve clinical symptoms, and alleviate the obstruction caused by bronchial neoplasm. Brouchoscopic argon plasma coagulation therapy is an effective and safe method for patients with bron-chial carcinoma.