CRISPR/Cas-mediated DNA base editing technology and its application in biomedicine and agriculture / 生物工程学报

In recent years, the genome editing technologies based on the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) have developed rapidly. The system can use homologous directed recombination (HDR) to achieve precise editing that it medicated, but the efficiency is extremely low, which limits its application in agriculture and biomedical fields. As an emerging genome editing technology, the CRISPR/Cas-mediated DNA base editing technologies can achieve targeted mutations of bases without generating double-strand breaks, and has higher editing efficiency and specificity compared with CRISPR/Cas-mediated HDR editing. At present, cytidine base editors (CBEs) that can mutate C to T, adenine base editors (ABEs) that can mutate A to G, and prime editors (PEs) that enable arbitrary base conversion and precise insertion and deletion of small fragments, have been developed. In addition, glycosylase base editors (GBEs) capable of transitioning from C to G and double base editors capable of editing both A and C simultaneously, have been developed. This review summarizes the development, advances, advantages and limitations of several DNA base editors. The successful applications of DNA base editing technology in biomedicine and agriculture, together with the prospects for further optimization and selection of DNA base editors, are discussed.

Knockdown the expression of ku70 and lig4 in HEK293T cells by CRISPR/Cas13 system / 生物工程学报

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.

Clinical Application of Etomidate Combined with Remifentanil by Target Controlled Infusion Anesthesia During Painless Bronchofiberscopy / 医药导报

Objective To discuss the safety of using etomidate combined with remifentanil by target controlled infusion ( TCI) for painless bronchofibroscopy. Methods Sixty patients were divided into two groups: painless bronchoscopy group (treatment group, 24 patients) and the routine bronchoscopy group (control group, 36 patients). Treatment group received TCI of remifentanil and intravenous injection of etomidate fat emulsion. Control group was subjected to surface anesthesia with 2%lidocaine. SpO2 , blood pressure, heart rate and breath changes during examination and complete awakening were continuously monitored. Bronchofiberscopy time, body movement during examination, bucking and satisfaction degree after examination were also recorded. Results Treatment group patients felt senseless and painless during bronchoscopy, without memory of bronchoscopy and pain. Patients in control group had discomfort, body movement and acute bucking, and most of them had painful memory. There were significant differences between the two groups (P0. 05). Conclusion TCI etomidate combined with remifentanil during bronchoscopy achieved satisfying anesthetic effect.

Modified purse-string suture in closure of skin defects: scar minimization and tension adjustment / 中华医学美学美容杂志

Objective To explore the effect of intraoperative tension adjustment and postoperative scar formation in the treatment of circular skin defects using modified purse-string suture.Methods Twenty-eight cases of circular lesions in the face region were selected and resected along the edge.We first used purse-string suture technique to reduce the circular defect area,and then closed the wound with linear suture in random direction.After 7 days,the stitches were removed and longterm follow-up effects were observed.Results All patients had no facial organ deformation and displacement,the length of the linear scar lesions was shorter than the preoperative length.In 28 cases,one patient's surgical wound was dehisced after removal of stitches in 24 h,and the rest of the skin defects healed after operation in stage Ⅰ.All cases were followed up for 3 months to 1 year,and had a satisfactory therapeutic effect.Conclusions Modified purse-string suture can linearly close the dicision in stage Ⅰ,reduce the scar area and maintain the normal morphology and relative position of the nearby organs.

Analysis of pedestrian road traffic crashes and injuries in Chongqing / 中华创伤杂志

Objeetive To analyze traffic accidents involving pedestrians in Chongqing from 2000 to 2006 so as to understand injury characteristics of the pedestrians. MethodsThe data of pedestrian road traffic crashes and injuries in three districts including one district in downtown,one in suburb and one in county of Chongqing from 2000 to 2006 were collected from road traffic crash and road traffic injury database to analyze severity of crashes,casualties,action and location of pedestrians in crashes. Results The road crashes led to 7 934 pedestrian injuries (24.74％) and 867 pedestrian deaths (49.46％).Intensity of the pedestrian road crashes in rural areas was higher than that in downtowns and suburbs.Of all pedestrian casualties,57.91％ were due to illegally crossing the driveway,and 15.43％ of the casualties were on the crosswalk.Most of the casualties occurred on the main roads,the third class highways and second main roads in Chongqing.But serious crashes occurred on the first and second class highways and substandard highways,which led to average 3.17,4.13 and 5.42 deaths,respectively.Pedestrians accounting for 52.46％ of the total severe injuries were more than the proportion among the minor to moderate injuries.Head injuries were the most common for the pedestrians and chest injuries were also usual among the severe injuries. Conclusions The intensity of pedestrian road crashes is higher than that of other crashes,especially in rural areas.It should be noticed that the main injuries of the pedestrians are head injuries,which is mainly resulted from illegally crossing driveway.It is worth noticing that a rather high proportion of pedestrians are impaired on the crosswalks,indicating that we should pay more attention to protect right of the pedestrians on the crosswalks during transportation law enforcement and safety education in the future.

Establishment of road traffic crash & traffic injury database / 中华创伤杂志

Objective To establish a road traffic crash (RTC) & traffic injury (RTI) database database with scientific and reasonable structure. Methods ( 1 ) A set of scientific and standard RTC&RTI survey form was made and a software for collection of data on RTC&RTI developed. (2) The general data on RTI&RTI of China was obtained from traffic management departments of China and the data of some districts in Chongqing from 2000 to 2006 were obtained. All the data were input into the database for analysis. Results A TRC&RTI database software V1. 0 was developed based on the survey form. The input items included data about the areas where the accidents occurred, the general situation of the specific accident, the road conditions, the accident causes and responsibilities, the vehicle and the injury to persons. We obtained the RTC and RTI data of China during past 13 years and collected data on 33 987 cases of RTC and 52369 RTI of four districts in Chongqing. The RTC&RTI database was established. Conclusions The database has involved detailed data of general road crashes and injury as well as individual crash, which provides a good support for further study of RTC and RTI.

Characteristics and risk factors of traffic injuries of adolescents (0-19 years old) in the urban and rural areas of Chongqing / 中华创伤杂志

ObjectiveTo study the characteristics and risk factors of traffic injuries among the adolescents (0-19 years old) in the urban and rural areas of Chongqing and discuss the preventive strategy. MethodsData of road traffic crashes and injuries involving 0-19 years old adolescents over a seven-year period (2000-2006) in three districts of Chongqing was picked up from Road _Traffic Crashes and Injuries Database. Epidemiological characteristics of the traffic crashes and injuries among 0-19 years old adolescents were analyzed and differences between rural and urban areas were studied. ResultsFrom 2000 to 2006, both incidence rate and mortality rate peaked in the urban fringe area were 95.43/100 000 and 3.44/100 000 respectively. Traffic accidents in the suburbs and counties had the highest injury extent, with mortality rate of 9.06％ and severe injury rate of 17.33％. Brain injuries were the primary cause of death. Among the victims, 62.7％ were the pedestrians, of which 20.72％ bore equal responsibility because of disobeying traffic signals, suddenly walking across roads and not walking along the sidewalk. Ninety percentage of the traffic accidents occurred in the sections with no traffic signals or traffic control system. ConclusionIt is important to enhance the traffic safety consciousness of teenagers and children, strengthen preventive measures in fringe areas and rural areas and improve the emergency medical system in rural areas to provide the teenagers and children with a safety traffic environment.

Establishment and efficacy evaluation of autoregression moving average model for prediction of general traffic accidents in one district of Chongqing / 中华创伤杂志

Objective To establish an autoregression moving average (ARMA) model for predicting general traffic accidents and analyzing distributional difference on time series and frequency of common traffic accident so as to provide certain basis for a prediction model with better stability and accuracy. Methods The data of road traffic accidents in one newly developed zone of Chongqing in 2000-2005 were collected. The monthly distribution regularity of road traffic accidents was analyzed with descriptive epidemiologic method. ARMA model was set up for retrospective and prospective prediction. The predicted data were compared. Results Based on the characteristics of monthly distribution, the frequency of general traffic accidents in this area showed a cyclic fashion. The frequency of general traffic accidents predicted by ARMA model had over 80% of coincidence with the actual value. Conclusion The ARMA model can be used to predict the frequency of general traffic accidents, with better accuracy of short-term prediction than the long-term prediction.

Development of Trauma Database System network version 3.0 / 中华创伤杂志

Objective To develop a trauma database and its network version which could adapt to the need of accumulation and analysis of trauma care data in hospitals.MethodsThe Trauma Data-base System was developed based on the basic data of trauma care course, with assistance of the Windows operation system and SQL SERVER 2000 database, with vb.net and asp.net as programming language and Browser/Server model as visiting model.Results The trauma database with normative 313 fields adaptive to the process and management of trauma in hospitals was established, and the network version of Trauma Database System V3.0 was programmed.Information stream of trauma care was established,which helped implementation of functions including relational selection between international classification of diseases of trauma and abbreviated injury scale, trauma scoring and trauma care quality evaluation, ar-bitrary query and report (including tri-crosstab).The system has been used in several hospitals and data collection of seismic patients in "5 ·12" Wenchuan earthquake.Conclusions A trauma database with a network version has been developed and the software suits to the trauma care data registration in hospital, which facilitates registration of the data on occurrence, diagnosis, treatment, outcome, quality evaluation of trauma and provides convenient query and analysis of trauma data.

Influence of katsutoxin and its extract on bone marrow colony-forming unit-granulocyte and monocyte following ~(60)Co gamma ray radiation / 中国组织工程研究

The experiment was performed at Basic Medical College and First Affiliated Hospital, Zhengzhou University from September 2004 to December 2005. Totally 60 Kunming mice were divided into 5 groups randomly: ①blank control group (n =15) and simple radiation group (n =15). The mice were given 0.2 mL sterile saline by intraperitoneal injection. ②antineoplastic polypeptide from Buthus Martensii Venom (APBMV) group (n =10) and APBMV plus radiation group (n =10) received 0.2 mL APBMV according to prepared concentration by intraperitoneal injection. ③Katsutoxin extract Ⅲ plus radiation group (n =10) received 0.2 mL katsutoxin extract Ⅲ by intraperitoneal injection every other 5.5 hours for 7 days. After 24 hours from the last injection, the mice were endured 60Co g ray radiation (80 cm, 7.5 Gy irradiation dose, 0.27 Gy/min dose rate). Then katsutoxin extract Ⅲ was given same as above for 7 days. Then bone marrow was extracted to be cultured to colony-forming unit-granulocyte and monocyte (CFU-GM). The findings showed that colony amount of APBMV plus radiation group and katsutoxin extract Ⅲ plus radiation group was obviously more than that of simple radiation group [(32?5),(27?3),(2?1)pieces/well,P

Development of Recognized Software on Multi-tumor Protein Chip Analysis System V1.0 / 医疗卫生装备

Objective To develop the system of tumor recognized tool software based on the examination results of multi- tumor protein chip. Methods On the basis of the diagnostic recognized equation, the examination results were comparison by HTA combining VB script and Javascript language program. Results The functions of the software include dates input of tumor protein chip,recognized results, dates interrogation and record printing of 10 kinds of common tumor. Conclusion The software can be diagnosed automatically with the sorts of tumor, which is a scientific, convenient and efficient tool for tumor clinician.