Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals ; Base Sequence ; Cloning, Organism ; Electrophoresis ; Endonucleases ; genetics ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; Gene Targeting ; methods ; Goats ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Zinc Fingers

Objeoctive To compare the effects of etomidate and propofol on cerebral oxygen metabolism in patients undergoing abdominal surgery.Methods Thirty-six ASA Ⅰ or Ⅱ patients aged 30-64 yr weighing 42-73 kg undergoing abdominal surgery under general anesthesia were randomly divided into 2 groups (n=18 each):group E etomidate and group P propofol.Left radial artery was cannulated for continuous direct BP monitoring.A catheter was inserted into right internal jugular vein(LJV) and advanced cephalad until jngatar bulb for blood sampling.Both groups received midazolam 0.08 ms/ks,fentanyl 3μg/kg and vecurunium 0.1 mg/kg and in addition group P received propofol 1.5 mg/ks and group E etomidate 0.3 mg/kg respectively for induction of anesthesia.Anesthesia was maintained with propofol infusion at 4-6 mg·kg-1·h-1 in group P and etomidate infusion at 0.4-0.7 mg·kg-1·h-1 in group E and intermittent iv boluses of fentanyl and vecuronium.ECG,MAP,HR,SpO2 and PET CO2 were continuously monitored.Blood samples were taken from radial artery and IJV for blood gas analysis and lactic acid measurement before induction of anesthesia(T1),immediately after intubation (T2),30 min after skin incision (T3) and at the end of operation(T4).The rate of cerebral O2 extraction (CERO2) was calculated.Results The hemodynamic variables were within the normal range throughout the anesthesia and operation.The oxygen saturation and oxygen partial pressure of both arterial and venous blood(SaO2,SjvO2,PaO2,PjvO2) rose significantly after induction of anesthesia in both groups.There was no significant difference in arterial and venous blood lactic acid level and SaO2,SjvO2,CaO2,CjvO2,Da-jvO2 or CERO2 at all time points between the two groups.Conclusion Both etomidate and propofol combined with midazolam and fentanyl can decrease cerebral O2 metabolic rate and there is no significant difference between the two groups.

Objective:To illustrate a relatively complete knowledge system (e.g., research outputs, current hotspots, and future trends) in the sepsis field and to help scholars grasp the scientific research direction or clinical focus of treatment.Methods:The relevant literatures of sepsis during the time from 1985 to 2019 in Web of Science database were collected. Sepsis-related research contents were generated using softwares (CiteSpace 5.6.R2 and VOSviewer 1.6.13), which using data mining, information processing and knowledge map methods, to analyze the historical evolution and predict the development trend.Results:A total of 8 189 papers on sepsis were published. The volume of publications were increasing yearly from 1985 to 2019, and reached the top list of 1 276 in 2019. For research contents of sepsis, it has formed the basic characteristics of sepsis which focusing on epidemiological studies and animal experiments. Through cluster analysis, the researches mainly focused on six aspects: septic rat, necrotizingenterocolitis, sepsis-associated encephalopathy, acute kidney injury (AKI), gut-derived sepsis, and inflammatory mediator. And it presented the literature characteristics that related to the injury or dysfunction of intestines, brain, liver, kidney or other organs, but the heart and lung researches were more marginal. Additionally, based on the top key words with the strongest citation bursts, it reflected that the development trend of the continuous attention hotspots with "endotoxin" or "endotoxin shock", the significant attention hotspots with "inflammation", "immunity" and "multiple organ dysfunction syndrome" (MODS), and the novel burst attention hotspots with sepsis management including "diagnosis" and "chemotherapy".Conclusions:Through the hotspots and trends visualization of sepsis, the current researches are prefer to animal experiments, epidemiology, or other basic scientific aspects. Meanwhile, the researches are mostly focusing on inflammatory reaction, immune function or organ dysfunctions. Integrating the knowledge maps of hotspots and trends, based on researches of epidemiology, diagnosis, risk factors, pathogenesis, or treatment, we predict that the future scientific topics will concentrating on childhood sepsis, organ injury mechanism or intervention relating to MODS, and integrated management of sepsis by combining traditional Chinese medicine and Western medicine.

Abstract: Objective To analyze the distribution characteristics of new occupational disease cases in Guangxi Zhuang “ ” Methods Autonomous Region (hereinafter referred to as Guangxi ) from 2016 to 2021. Through the Occupational Disease Report Card of Occupational Disease and Occupational Health Monitoring Information System, a subsystem of China Disease Prevention and Control Information System, the data of occupational disease reported in Guangxi from 2016 to 2021 were Results collected and analyzed by routine data analytic method. A total of 633 new cases of occupational diseases were diagnosed in Guangxi from 2016 to 2021. Most of the cases occurred in males that account for 96.5% (611/633). Among them, 85.8% of cases were occupational pneumoconiosis, 6.3% occupational otoaryngological and stomaological diseases, 3.0% chemical poisoning and 4.9% other five types of occupational diseases. The geographical distribution was dominated in Hechi Citythataccountsfor51.7%.Theindustrialdistributionwasconcentratedinbituminouscoalminingandwashing,tinminingand dressing, lead and zinc mining and dressing (43.1% of the total). Private enterprises account for 47.3%. The enterprise was - mainlysmall sizedenterprises,accountingfor50.0%.Themaintypesofworkwererockdrillsandmaincoalminer,accountingConclusion for18.8%and17.5%,respectively. Occupationalpneumoconiosiswasthemostimportantoccupationaldiseasein Guangxi. It is necessary to strengthen the occupational hazard exposure control and protection of bituminous coal mining and - - washing,tinmininganddressing,leadandzincmininganddressingindustries,privateenterprises,andsmall andmedium sized enterprises,rockdrillsandmaincoalminerinHechiCity.

We compared the efficiency of cloning goat using human lactoferrin (hLF) with genetically modified donor cells marked by single (Neo(r)) or double (Neo(r)/GFP) markers. Single marker expression vector (pBLC14) or dual markers expression vector (pAPLM) was delivered to goat fetal fibroblasts (GFF), and then the transgenic GFF was used as donor cells to produce transgenic goats. Respectively, 58.8% (20/34) and 86.7% (26/30) resistant cell lines confirmed the transgenic integration by PCR. Moreover, pAPLM cells lines were subcultured with several passages, only 20% (6/30) cell lines was observed fluorescence from each cell during the cell passage. Somatic cell nuclear transfer using the donor cells harbouring pBLC14 or pAPLM construct, resulting in a total of 806 reconstructed embryos, a pregnancy rate at 35 d (53.8%, 39.1%) and 60 d (26.9%, 21.7%), and an offspring birth rate (1.9%, 1.4%) with 5 and 7 newborn cloned goats, respectively. Transgene was confirmed by PCR and southern-blot in all cloned offspring. There were no significant differences at the reconstructed embryo fusion rates, pregnancy rates and the birth rate (P > 0.05) between single and double markers groups. The Neo(r)/GFP double markers could improve the reliability for accurately and efficiently selecting the genetically modified donor cells. No adverse effect was observed on the efficiency of transgenic goat production by SCNT using somatic cells transfected with double (Neo(r)/GFP) markers vector.
Animals ; Animals, Genetically Modified ; genetics ; Cloning, Molecular ; Cloning, Organism ; methods ; veterinary ; Fetus ; Fibroblasts ; cytology ; Genetic Markers ; Goats ; embryology ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Lactoferrin ; biosynthesis ; genetics ; Neomycin ; Nuclear Transfer Techniques ; veterinary ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; veterinary

Adult ; Air Pollutants, Occupational ; analysis ; Dust ; analysis ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Occupational Health ; Welding ; Young Adult

Objective To examine the influence of systemic rehabilitation exercise on social functioning in chronic schizophrenic patients.Methods 60 qualified cases selected from chronic psychiatric inpatient department were randomly divided into two groups according to admission number (AD).30 cases in treatment group accepted hospitalized systemic rehabilitation exercise and 30 controls accepted ordinary treatment for 6 months.Social functio-ning evaluated with SSPI and PSP.Results The total score of SSPI(t=1.322,P=0.256) and PSP(t=1.563,P=0.362) were not significantly higher(P>0.05),The total score of SSPI [(37.44 ±4.33)points,t=2.719,P=0.001]and PSP[(72.14 ±6.86)points,t=3.985,P=0.000]total score in the treatment group were significantly higher than that in the control group(P<0.01).It were significantly higher in the treatment group than before the intervention.The total score of SSPI (t=0.385,P=0.763)and PSP(t=0.682,P=0.827) total score were not significantly higher in the control group(P>0.05).Conclusion Systemic rehabilitation training does influence on social functioning in chronic schizophrenic patients.

Objective To study the clinical curative effect of syndrome differentiation-based treatment with traditional Chinese medicine on patient with Chronic Kidney Disease (CKD3-4). Method According to prospective multicenter randomized controlled study which was applied to the patients with CKD3-4 , the total of 339 patients were divided into two groups. To one group as standard including 167 patients ,the other including 172 patients. Chinese Traditional Patent Medicine single or combined-prescription oral treatment according to syndrome differentiation-based treatment was employed. To the other group as reference , Chinese Traditional Patent Medicine oral treatment according to experienced rule was applied. The clinical curative effect results were obtained after a 24 weeks-treatment. Result The Scr ,eGFR and UTP with CKD3 in standard group illustrate statistics significance (P<0.05). On the opposite, those of the reference group remain insignificance (P > 0.05). The clinical curative effect result of patient in standard group was better than that of reference group. About the clinical curative effect of CKD4, no statistics significance difference (P > 0.05) could be found between groups or between patients in one group. The TCM score of patient with CKD3-4 in standard group have improved (P < 0.05) according to baseline. However, that of reference group did not show any statistics significance (P > 0.05). Conclusion Syndrome differentiation-based treatment with traditional Chinese medicine illustrated a better result in protection of renal function, reduction in urinary protein and decline in TCM score than the treatment according to experienced rule.

MicroRNA (miRNA or miR) is a class of highly conserved endogenous non-coding RNA of 21~25nt, which is widely existed in organisms. Currently, miRNA has been proven to be associated with cardiovascular diseases in clinical research, but it has not been reported in the field of forensic medicine. This paper highlights recent findings about miRNA and its application in cardiovascular diseases, and the application aspect of miRNA in sudden cardiac death in forensic science.

Objective:To investigate the effect of abnormal D-dimer level on the prognosis of patients with frontal lobe injury, and to provide a new idea for clinical diagnosis, treatment and prognosis evaluation of frontal lobe injury.Methods:A retrospective analysis was performed on 71 patients with traumatic brain injury (TBI) who were admitted to the Surgical Care Unit of the First Affiliated Hospital of Zhejiang University School of Medicine from November 1, 2020 to February 1, 2021. According to whether TBI involved in the frontal lobe, the patients were divided into the experimental group ( n = 44) and the control group ( n = 27). Clinical data including sex, age, body mass index (BMI), length of hospital stay, ISS trauma score, GCS coma score, qSOFA score, GOS prognostic score and coagulation index test results within 24 h of admission of the two groups were recorded, and the difference of each index between the two groups was compared. Results:① D-dimer levels were significantly higher in the experimental group than in the control group ( P<0.01); ② The incidence of complications in the experimental group was 40.9%, of which the incidence of lower limb vein thrombosis was 18.2%, and the incidence of complications in the control group was 18.5%, of which the incidence of lower limb vein thrombosis was 3.7%; there were statistical differences between the two groups ( P<0.05); ③ The average hospital stay of patients was 10.96 days in the control group, and 15.50 days in the experimental group with a statistically significant difference between the two groups ( P<0.05); ④ The 28-day mortality rate of patients in the experimental group was 18.2% and 7.4% in the control group. There was no significant difference in 28-day mortality between the two groups ( P>0.05); ⑤ The level of D-dimer was negatively correlated with GOS prognostic score in patients with frontal lobe injury ( r=-0.501, P<0.01). Conclusions:Patients with frontal lobe injury have increased D-dimer levels, increased incidence of complications, and longer hospital stay. There is a significant negative correlation between D-dimer level and prognosis in patients with frontal lobe injury. D-dimer can be used as a sensitive indicator to evaluate the prognosis of patients with frontal lobe injury. The higher the D-dimer level, the worse the prognosis of patients.