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

There are many kinds o f Taraxacum plants，which are widely distributed . The dried whole plants of many kinds of plants can be used as medicinal materials ，and the origin is confused . This study finds that Taraxaci Herba first appears as “Pugoncao”in the Liu Juanzi Guiyifang of Jin Dynasty after consulting ancient herbal books ，medical books ，prescription books and modern literature . According to characteristics ，morphology，color and sound of the plant ，it has “Huanghualang”， “Gounoucao”，“Fugongying”，“Pugongying”，“Huanghuadiding”and other aliases . Taraxaci Herba originates from the dried whole plant of Compositae family Taraxacum mongolicum Hand.-Mazz.，T. borealisinense Kitam. or several plants of the same genus . Similar to it ，Sonchus oleraceus Linn.（also known as “Kujucai”）cannot replace Taraxaci Herba as medicine . Because of its characteristics of rapid reproduction and vigorous growth ，Taraxaci Herba is widely distributed in most areas of China .“Absolutely no in Lingnan ”stated in Bencao Gangmu is inconsistent with the actual situation after consulting the relevant works . Ancient physicians take growth height of Taraxaci Herba as their quality standard . Now，those with many leaves ，gray green color and long roots are preferred . Ancient doctors pointed out that Taraxaci Herba were mostly harvested in April and May of the lunar calendar ， and modern research believes that it should be harvested in the near flowering period . In ancient times ，Taraxaci Herba is processed by many methods ，such as purification ，charcoal-frying，honey-frying and wine -frying. In modern times ，purification is the main method. This paper provides theoretical support for the value development and rational utilization of Taraxaci Herba by systematically researching its name ，origin，place of production ，harvesting and processing .

Cell pyroptosis is a programmed death of inflammatory cells. Many members of the gasdermin family （the effector protein family that forms pores） participate in the pathological process of a variety of diseases， such as cancer， myocardial ischemia， renal injury， and osteoarthritis， mainly by activating cysteine aspartate-specific protease （Caspase） for polymerization and shear. Cell pyroptosis has bidirectional regulation. Induction of pyroptosis can promote cell clearance under pathological conditions （such as cancer and tumor cells）， but long-term induction of pyroptosis can lead to abnormal lipid and related vitamin metabolism in vivo. Regulating the balance between cell pyroptosis and proliferation is an important target for traditional Chinese medicine （TCM） treatment of diseases. The Yin-Yang theory runs through the whole process of TCM diagnosis and treatment， which is used to explain the physiological and pathological changes of human body and guide the theory， diagnosis and treatment of diseases and health care. The balance between cell proliferation and pyroptosis is essentially the embodiment of Yin-Yang balance at the cellular level， and the theory of Yin-Yang spontaneous harmonization dominates the balance. TCM intervention on cell pyroptosis is mainly reflected in promoting and inhibiting cell pyroptosis， which has the same significance as Yin-Yang regulation. Based on this theory， this paper revealed the relationship between TCM inhibiting and promoting cell pyroptosis through the Yin-Yang theory， to provide theoretical support for the modernization of the Yin-Yang theory and new goals for the diagnosis and treatment of diseases.

Objective:To explore the clinical efficacy of microsurgical technique combined with cosmetic suture technique in situ replantation of total scalp avulsion.Methods:The clinical data of patients with total scalp avulsion admitted to the Department of Burn and Plastic Surgery of General Hospital of Northern Theatre Command from January 2015 to December 2020 were collected. All patients were treated by microsurgical technique combined with cosmetic suture technique. Thorough debridement was performed during the operation. Active anti-shock treatment was given to the patient with hemorrhagic shock. The vascular was explored and labeled. The vascular anastomosis was conducted by microsurgical technique. The venous anastomosis was performed in the first place. The arterial anastomosis was performed according to the patient's condition. Symptomatic treatment, such as routine anti-coagulation, was given to patients after the operation. An infection prevention strategy was performed in patients. The recovery of patients was observed.Results:A total of six patients were involved in this study. All patients were females, aged from 36 to 55 years old. Various degrees of wound contamination can be seen, and one patient with hemorrhagic shock. The superficial temporal vein, supraorbital vein and sentinel vein were anastomosed in two patients, and the superficial temporal vein and sentinel vein were anastomosed in four patients. The superficial temporal artery and supratrochlear artery were anastomosed in two patients, and the superficial temporal artery was anastomosed in four patients. All patients were followed up for 3 months to 2 years, averaging 13.71 months. More than 90% of the scalp survived in three patients, 80% in two patients, and 50% in one patient. The necrotic area survived after replantation of a split-thickness skin graft. The sensory function of surviving scalp after replantation was slightly regressed. The hair growth was normal. Only superficial linear scar left. All patients were satisfied with the efficacy.Conclusions:The application of the microsurgical technique for vascular anastomosis combined with cosmetic suture technique for repair can achieve good clinical efficacy in situ replantation of total scalp avulsion.

Objective:To explore the clinical efficacy of microsurgical technique combined with cosmetic suture technique in situ replantation of total scalp avulsion.Methods:The clinical data of patients with total scalp avulsion admitted to the Department of Burn and Plastic Surgery of General Hospital of Northern Theatre Command from January 2015 to December 2020 were collected. All patients were treated by microsurgical technique combined with cosmetic suture technique. Thorough debridement was performed during the operation. Active anti-shock treatment was given to the patient with hemorrhagic shock. The vascular was explored and labeled. The vascular anastomosis was conducted by microsurgical technique. The venous anastomosis was performed in the first place. The arterial anastomosis was performed according to the patient's condition. Symptomatic treatment, such as routine anti-coagulation, was given to patients after the operation. An infection prevention strategy was performed in patients. The recovery of patients was observed.Results:A total of six patients were involved in this study. All patients were females, aged from 36 to 55 years old. Various degrees of wound contamination can be seen, and one patient with hemorrhagic shock. The superficial temporal vein, supraorbital vein and sentinel vein were anastomosed in two patients, and the superficial temporal vein and sentinel vein were anastomosed in four patients. The superficial temporal artery and supratrochlear artery were anastomosed in two patients, and the superficial temporal artery was anastomosed in four patients. All patients were followed up for 3 months to 2 years, averaging 13.71 months. More than 90% of the scalp survived in three patients, 80% in two patients, and 50% in one patient. The necrotic area survived after replantation of a split-thickness skin graft. The sensory function of surviving scalp after replantation was slightly regressed. The hair growth was normal. Only superficial linear scar left. All patients were satisfied with the efficacy.Conclusions:The application of the microsurgical technique for vascular anastomosis combined with cosmetic suture technique for repair can achieve good clinical efficacy in situ replantation of total scalp avulsion.

Approximately 15% of couples suffer from infertility worldwide, and male factors contribute to about 30% of total sterility cases. However, there is little progress in treatments due to the obscured understanding of underlying mechanisms. Recently microRNAs have emerged as a key player in the process of spermatogenesis. Expression profiling of miR-181a was carried out in murine testes and spermatocyte culture system. In vitro cellular and biochemical assays were used to examine the effect of miR-181a and identify its target S6K1, as well as elucidate the function with chemical inhibitor of S6K1. Human testis samples analysis was employed to validate the findings. miR-181a level was upregulated during mouse spermatogenesis and knockdown of miR-181a attenuated the cell proliferation and G1/S arrest and increased the level of S6K1, which was identified as a downstream target of miR-181a. Overexpression of S6K1 also led to growth arrest of spermatocytes while inhibitor of S6K1 rescued the miR-181a knockdown-mediated cell proliferation defect. In human testis samples of azoospermia patients, low level of miR-181a was correlated with defects in the spermatogenic process. miR-181a is identified as a new regulator and high level of miR-181a contributes to spermatogenesis via targeting S6K1.

Objective:To investigate the prognostic factors for long-term outcomes in elderly patients after colorectal cancer(CRC)resection.Methods:Patients(aged ≥60 years)undergone CRC resection in 2004 with more than 10-year survival records from the SEER database were retrospective analyzed.Potential prognostic risk factors for survival, such as age, sex, race, marital status, tumor site and size, clinical staging, TNM staging and lymph node clearance(lymph nodes dissection)were assessed by using the Cox proportional hazards model.Results:A total of 15 775 CRC patients were enrolled in this study, including 7 828(49.6%)males and 7 947(50.4%)females.The 10-year overall survival(OS)was shorter in patients aged over 80 years than in patients aged 60 years( P=0.000). Multivariate analysis showed that females had a better 10-year OS than males( P=0.000). Univariate analysis showed no difference in 10-year OS between females and males( P=0.837). Compared with married patients, single, unmarried or widowed patients displayed a poorer 10-year OS( P=0.000)and that patients with cecum and appendix cancer had the worst 10-year OS( P=0.000), but it did not find any difference between primary tumors in different sites.Patients with a tumor size over 5 cm or at advanced stage(T3-T4, N1-N2 and M1)had a low 10-year OS( P=0.000). Conclusions:Old age, black race, single, unmarried or widowed status, tumor size over 5 cm and advanced stage(T3-T4, N1-N2 and M1)are negative predictors for 10-year OS in CRC patients and these findings can be used for education and treatment improvement strategies for CRC in future.

Approximately 15% of couples suffer from infertility worldwide, and male factors contribute to about 30% of total sterility cases. However, there is little progress in treatments due to the obscured understanding of underlying mechanisms. Recently microRNAs have emerged as a key player in the process of spermatogenesis. Expression profiling of miR-181a was carried out in murine testes and spermatocyte culture system. In vitro cellular and biochemical assays were used to examine the effect of miR-181a and identify its target S6K1, as well as elucidate the function with chemical inhibitor of S6K1. Human testis samples analysis was employed to validate the findings. miR-181a level was upregulated during mouse spermatogenesis and knockdown of miR-181a attenuated the cell proliferation and G1/S arrest and increased the level of S6K1, which was identified as a downstream target of miR-181a. Overexpression of S6K1 also led to growth arrest of spermatocytes while inhibitor of S6K1 rescued the miR-181a knockdown-mediated cell proliferation defect. In human testis samples of azoospermia patients, low level of miR-181a was correlated with defects in the spermatogenic process. miR-181a is identified as a new regulator and high level of miR-181a contributes to spermatogenesis via targeting S6K1.

Objective:Tumor-treating fields (TTFields) is a kind of non-invasive anti-mitotic tumor therapy, which has been approved for patients with newly diagnosed and recurrent glioblastoma. This study aims to explore the efficacy and safety of TTFields in high-grade gliomas in clinical practice settings.Methods:The clinical data of 15 patients with recurrent glioma and 9 patients with newly diagnosed high-grade glioma admitted to our center from April 2019 to January 2021 were retrospectively analyzed. All patients accepted TTFields≥1 month. Follow-up was performed for 5.3 months (ranged from 2.3 to 10.7 months); Response Assessment in Neuro-Oncology Working Group (RANO) criteria was used to evaluate the glioma responses. The progression-free survival (PFS) and overall survival (OS) were calculated according to Kaplan-Meier method. Common Terminology Criteria for Adverse Events v5.0 (CTCAE v5.0) and TTFields related skin adverse reaction (dAE) criteria were used to evaluate the adverse events. Quality of life questionnaire-core 30 (QLQ-C30) and QLQ-brain cancer module (QLQ-BN20) questionnaires were used to evaluate the health-related quality of life (HRQoL). Treatment compliance was evaluated by data on the use of NovoTTF-200A devices, and calculated as a percentage of daily TTFields usage.Results:The median duration of TTFields was 4.2 months (ranged from 1.0 to 10.7 months), with a median compliance rate of 91.5% (67.0%-97.0%). TTFields was used alone in 2 patients and used with combination of chemotherapy in 22 patients. From follow-up to April 2021, 14 patients had stable symptoms and 10 had disease progression (8 died). The median PFS and OS of recurrent patients were 5.9 months ( 95%CI: 3.3-8.6 months) and 8.5 months ( 95%CI: 3.2-13.8 months), respectively; and the median PFS and OS of newly diagnosed patients were both 10.7 months (without 95%CI). The common adverse events included grading 1 dAE (58.3%) and grading 2 dAE (12.5%), without grading 3 or 4 dAE, manifested as contact or allergic dermatitis, erosion, folliculitis and ulcers. And 87.5% patients had stable HRQoL. Conclusions:The preliminary results showed that the survival of recurrent high-grade glioma patients treated by TTFields is similar to that reported in foreign literature; and the newly diagnosed patients need further survival follow-up. The patients' treatment compliance and safety are good. The dAE incidence (grading 1-2) is higher than that reported in the literature, and the toxicity was acceptable.

With the development of fine anatomy,the concept of mesopancreas has been proposed,and total mesopancreatic excision continues to be used in clinical practice.This method has become the best surgical method of pancreatic head cancer.But due to deep position of pancreas and mesopancreas,the surrounding adjacent organs and complexs tructure and other factors,experts are concerned about at the presence or absence of the mesopancreas.So the scope of clinical surgical resection structural reservations are also controversial.This article reviewed the development of embryonic development of the pancreas and the research progress in order to provide theoretical support for the study of the fine anatomy of the pancreas and the precise operation of pancreatic diseases.