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

ObjectiveTo evaluate the predictive value of the sperm quality to fertilization outcomes after short-time insemination.MethodsA total of 558 cycles of short-time insemination in the Reproductive Medical Center of Henan Provincial People's Hospital during January 2009 to June 2010 excluding patients aged ＞ 38 years and M Ⅱ oocyte number ＜ 3 were analyzed retrospectively.According to whether undergo rescue intracytoplasmic sperm injection( Re-ICSI),all cycles were divided into in vitro fertilization (IVF)group (472 cycles) and rescue intracytoplasmic sperm injection (Re-ICSI) group (86 cycles).Both IVFgroup and Re-ICSI group were subdivided into primary infertility and secondary infertility according to previous history of pregnancy.269 primary infertility cycles and 203 secondary infertility cycles were characterized in IVF group; and 64 primary infertility cycles and 22 secondary infertility cycles were characterized in Re-ICSI group.x2 test was applied for comparison of embryo plant rate,clinical pregnancy rate,early miscarriage rate between IVF and Re-ICSI groups,while Fisher test was used for comparison of live birth rate.and Mann-Whitney U test was utilized for comparison of duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity between IVF and Re-ICSI groups.ResultsThe embryo plant rate,clinical pregnancy rate,early miscarriage rate,live birth rate of IVF group were:29.4％,44.9％,13.4％,37.0％ respectively; the above indicators in Re-ICSI group were:25.7％,34.6％,10.7％,29.6％ respectively,the differences of the indicators between the two groups had no statisticalsigmficance (x2 =0.869,2.963,0.010,P =0.351,0.085,0.922,0.098).Median of duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity of primary infertility cycles in IVF group were:4.00(3.00 -6.00) years,58.37(33.64 - 102.27) × 106,81.09％ (79.41％ -88.69％ ),76.30 (48.50 - 92.46 ) μIU/106 sperm respectively ; in Re-ICSI group were:5.00 ( 3.25 -8.00) years,36.33 (20.59 -64.43 ) × 106,85.5％ (81.28％ - 89.02％ ),47.14( 31.61 -90.24) μIU/106 sperm respectively,the differences of them between the two groups had statistical significance (Z =-2.617, -3.505, -3.553, -3.530,P =0.009,0.000,0.000,0.000).Median of duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity of secondary infertility cycles in IVF group were:5.00 (3.00 -7.00) years,63.00 (34.20 - 107.73 ) × 106,81.29％ (79.90 -86.09) ％,78.34 ( 53.87 - 98.00) μIU/106 sperm respectively,in Re-ICSI group were:5.00 ( 3.75 -7.00) years,28.80 ( 18.57 - 48.56 ) × 106,88.79％ ( 84.04 - 95.64 ) ％,54.70 ( 39.73 - 76.77 ) μIU/106 sperm respectively,the differences of them between the two groups showed statistical significance except duration of infertility (Z =- 0.338,- 3.505,- 3.553,- 3.530,P =0.735,0.000,0.000,0.006).ConclusionThe duration of infertility,forward moving sperm counts,abnormal sperm rate,sperm acrosin activity have predictive value of fertilization outcomes after short-time insemination.

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.

Objective To investigate the status of cognitive impairment and its associated factors among the elderly in community in Beijing. Methods 2055 elderly people aged 60 or more from communities in Beijing were investigated and assessed with Mini-Mental State Examination (MMSE). Results 1985 from 2055 met the requirements of the questionnaire. Univariate analysis showed that age (F=37.068,P<0.001), gender (β=-0.187), educational level (F=21.875, P<0.001), marriage (β=0.328), social communication (β=0.124), and frequency of physical exercise (F=21.58, P=0.01, β=0.126) were related to the scores of MMSE (P<0.001). While multivariate analysis showed that educational level (β=0.421), age (β= -0.226), gender (β=-0.226), and frequency of physical exercise (β=0.128) were possibly the influential factors for the scores of MMSE (P<0.001). Conclusion Age, educational level, and frequency of physical exercise are important factors affecting cognitive functions of the elderly, in which age is negative, and the others are positive.

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 investigate the diagnostic value of Astograph methacholine provocation test in patients with chest tightness variant asthma ( CTVA)??Methods From January 2011 to February 2017,156 patients with CTVA in outpatient or inpatient department of respiratory medicine of Kailuan General Hospital affiliated to North China University of Science and Technology were selected as case group ( chest tightness variant asthma group )??The control group were 361 non?asthmatic patients including interstitial lung disease ( 23 cases), coronary disease ( 157 cases), hypertensive cardiopathy ( 22 cases), myocardiosis (16 cases),congenital heart disease ( 3 cases),rheumatic valvular heart disease (6 cases), central airway disease (3 cases),thyromegaly (10 cases),mediastinal tumor (5 cases),thoracic or spinal deformity (8 cases),phrenoparalysis (2 cases) and vegetative nerve functional disturbance (106 cases)??All participants received pulmonay ventilation test, average daily and nightly variation rate of PEF ( Peak expiratory flow) or PEF weekly variability, Astograph methacholine provocation test ( forced expirataory volume in one second≥70% expectation),and other relevant examinations??The diagnostic value of Astograph methacholine provocation test on CTVA was assessed by analyzing the sensitivity, specificity, positive predictive value,negative predictive value,and Yunden index of Astograph methacholine airway??Results Compared with the control group (( 1??18 ± 0??44)%), theforced expiratory flow from 75% of Forced vital capcacity ( FEF75 ) index of CTVA group (( 1??29 ± 0??50 )%) had significant difference (, t＝ 2??96, P＝0??006)??The sensitivity,specificity,positive predictive value,negative predictive value,Yunden index,and diagnostic accuracy of Astograph methacholine provocation test on CTVA were 0??814,0??695,0??536,0??305, 0??509 and 0??731, respectively??Conclusion The sensitivity, negative predictive value, Yunden index and diagnostic accuracy of Astograph methacholine provocation test on CTVA were higher,whereas the specificity and positive predictive value were relatively lower,suggesting that Astograph methacholine provocation test had a reliable diagnostic value on CTVA,with lower false negative and higher false positive??

The contents recorded in the rotation registration manual is not only the quantitative indicators for evaluating the quality of residency training, but also the important basis for training assessment and issuance of training certificates. In order to solve the problems of data authenticity, information delay, and repeated entry in the rotation registration manual for residency training, Shanghai East Hospital, Tongji University, launched a project to dock the electronic rotation registration manual with the hospital information system. Through the establishment of the project team, the development of working mechanisms, and the implementation of the project, data analysis was used for process reformation and system optimization, so as to continuously improve management efficiency and medical safety while solving problems and form a set of implementation system with reference significance in practice.

Objective:To explore the application effect of parent-child emotional regulation and resilience group training in adolescent depression patients.Methods:From August 2020 to September 2021, a total of 118 adolescent depression patients were enrolled and randomly divided into the intervention group(66 cases) and the control group(66 cases) by a random number table method.The intervention group received medication therapy and parent-child group emotional regulation and psychological resilience training, while the control group received medication therapy and commonly used individual, family or group therapy in clinical practice.The Hamilton Depression rating scale(HAMD-24 version), Herth hope scale(HHS), Connor-Davidson resilience scale(CD-RISC), and family adaptability and cohesion evaluation scale, second edition Chinese version(FACES-Ⅱ-CV) were adopted to investigate participants at baseline, 12-week, 24-week, and 36-week after intervention.SPSS 26.0 statistical software was used to perform repeated measurement analysis of variance on the data.Results:(1)The interaction effect between two groups of HAMD scores( F=54.0, P<0.001), group main effect( F=401.4, P<0.001), and time main effect( F=116.6, P<0.001) were all significant.Further simple effect analysis showed that there were statistically significant differences in HAMD scores at various time points after intervention between the intervention group(26.2±6.5, 19.3±5.9, 11.3±5.6) and the control group(33.1±9.1, 30.3±7.9, 25.0±8.4)(all P<0.05). Intragroup comparison showed the HAMD scores of the intervention group and control group at each time point after intervention were lower than those before intervention (all P<0.05). (2)The interaction effect of CD-RISC scores between two groups of patients( F=72.1, P<0.001), group main effect( F=48.9, P<0.001), and time main effect( F=174.9, P<0.001) were significant.Further simple effect analysis showed that the CD-RISC score of the intervention group at each time point after intervention were higher than those of the control group(all P<0.05). Intragroup comparison showed the scores of CD-RISC at each time point after intervention in the intervention group and the control group were higher than those before intervention(all P<0.05). (3)The interaction effect of HHS scores( F=121.6, P<0.001), group main effect( F=57.4, P<0.001), and time main effect( F=208.1, P<0.001) of the two groups of patients were significant.Further simple effect analysis showed that the HHS scores of the intervention group were higher than those of the control group at all time points after intervention(all P<0.05). Intragroup comparison showed the HHS scores of the intervention group and the control group at each time point after intervention were higher than those before intervention(all P<0.05). (4)The interaction effect of FACES-Ⅱ-CV scores( F=45.0, P<0.001), group main effect( F=20.3, P<0.001), and time main effect( F=154.5, P<0.001) of the two groups of patients were significant.Further simple effect analysis showed that the FACES-Ⅱ-CV scores of the intervention group were higher than those of the control group at all time points after intervention(all P<0.05). Intragroup comparison showed the FACES-Ⅱ-CV scores of the intervention group and the control group at each time point after intervention were higher than those before intervention(all P<0.05). (5)The total effective rate of the intervention group was higher than that of the control group(95.1%, 87.7%)( P<0.001). Conclusion:In adolescents with depression, parent-child group emotional regulation and resilience training can effectively reduce depression emotion, increase the level of hope and resilience of patients and enhance family intimacy and adaptability.

OBJECTIVETo assess the levels of polycyclic aromatic hydrocarbons monohydroxy metabolites in urine of general population in China among 8 provinces, provide the baseline of the metabolites in the general population.

METHODSFrom 2009 to 2010, 18 120 subjects of general population aged 6-60 years old were recruited from 24 areas among 8 provinces in east, west and central areas of China mainland by cluster random sampling. The information of the living environment and health condition were collected by questionnaire and spot urine samples were collected, 4 680 urine samples were analysed by high performance liquid chromatography with tandem mass spectrometry, and monohydroxy metabolites distribution in urine among groups of gender and ages were analysed.

RESULTSGeometric means (GM) of 2-naphthol, 1-naphthol, 3-phenanthrol and 1-hydroxypyrene concentration in urine (95%CI) were 1.85 (1.75-1.95), 1.55 (1.50-1.61), 0.57 (0.54-0.59) and 0.82 (0.78-0.85) µg/L, respectively;and median are 2.44, <0.50, 0.72 and 0.90 µg/L, respectively. The concentration between male and female were significantly different (P < 0.01), and the concentration among the groups of population were significantly different (P < 0.01), the GM of 2-naphthol among the groups of population aged 6-12, 13-16, 17-20, 21-30, 31-45 and 46-60 years old were 1.60, 1.56, 1.69, 2.23, 1.91 and 1.86 µg/L (χ(2) = 17.90, P < 0.01), the GM of 1-naphthol in the groups were 1.30, 1.16, 1.53, 1.68, 1.80 and 1.52 µg/L (χ(2) = 76.22, P < 0.01), the GM of 3-phenanthrol in the groups were 0.78, 0.76, 0.55, 0.42, 0.50 and 0.99 µg/L (χ(2) = 66.48, P < 0.01), the GM of 1-hydroxypyrene in the groups were 0.77,0.64, 1.00, 0.84, 0.84 and 0.57 µg/L (χ(2) = 51.48, P < 0.01), respectively.

CONCLUSIONThe distribution of monohydroxy metabolites levels in urine of general population were different, it provided a basic data for the further study of polycyclic aromatic hydrocarbons biomonitoring in the population.

Adolescent ; Adult ; Child ; China ; epidemiology ; Cross-Sectional Studies ; Environmental Exposure ; Female ; Humans ; Male ; Middle Aged ; Naphthols ; urine ; Polycyclic Aromatic Hydrocarbons ; urine ; Pyrenes ; urine ; Sentinel Surveillance ; Young Adult

COVID-19 ; Humans ; Inflammation/genetics* ; Lung ; Pneumonia ; SARS-CoV-2 ; Serologic Tests