1.Establishment and optimization of sequence-related amplified polymorphism amplification system for Carthamus tinctorius L.
Academic Journal of Second Military Medical University 1985;0(05):-
Objective:To study the factors influencing the amplification of sequence-related amplified polymorphism(SRAP) system in Carthamus tinctorius L.and to establish a stable SRAP reaction system,laying a foundation for molecular marker assistant breeding of Carthamus tinctorius L.Methods: Cetyltrimethylammonium bromide method was used to extract the genomic DNA of Carthamus tinctorius L..Twenty-seven tests with 3 factors at 3 levels were designed,the conditions including Taq polymerase concentrations(0.02,0.04,0.06 U/?l),dNTP concentrations(0.15,0.25,0.30 mmol/L)and Primer concentrations(0.15,0.30,0.45 ?mol/L);another 8 tests were designed based on the single factor of Mg~(2+) concentrations(0.5,1.0,1.5,2.0,2.5,3.0,3.5 and 4.0 mmol/L).Twenty ng DNA template was added into 25 ?l SRAP reaction system;the system was optimized and agarose electrophoresis was used for determination.Results: A SRAP reaction system for Carthamus tinctorius L.was established.In a 25 ?l reaction system,Taq polymerase was 0.02 U/?l,dNTP was 0.25 mmol/L,Primer was 0.30 ?mol/L,and Mg~(2+) was 3.0 mmol/L.Target bands increased after optimization,with good reproducibility,and the amplification results were satisfactory.Conclusion: The SRAP reaction system in this experiment is suitable for analysis of Carthamus tinctorius L.
2. Establishment and optimization of sequence-related amplified polymorphism amplification system for Carthamus tinctorius L
Academic Journal of Second Military Medical University 2006;27(5):544-547
Objective: To study the factors influencing the amplification of sequence-related amplified polymorphism (SRAP) system in Carthamus tinctorius L. and to establish a stable SRAP reaction system, laying a foundation for molecular marker assistant breeding of Carthamus tinctorius L. Methods: Cetyltrimethylammonium bromide method was used to extract the genomic DNA of Carthamus tinctorius L. Twenty-seven tests with 3 factors at 3 levels were designed, the conditions including Taq polymerase concentrations (0.02, 0.04, 0.06 U/μl). dNTP concentrations (0.15, 0.25, 0.30 mmol/L) and Primer concentrations (0.15, 0.30, 0.45 μmol/L); another 8 tests were designed based on the single factor of Mg2+ concentrations (0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5 and 4.0 mmol/ L). Twenty ng DNA template was added into 25 μl SRAP reaction system; the system was optimized and agarose electrophoresis was used for determination. Results: A SRAP reaction system for Carthamus tinctorius L. was established. In a 25 μl reaction system, Taq polymerase was 0.02 U /μl, dNTP was 0.25 mmol/L, Primer was 0.30 μmol/L, and Mg2+ was 3.0 mmol/ L. Target bands increased after optimization, with good reproducibility, and the amplification results were satisfactory. Conclusion: The SRAP reaction system in this experiment is suitable for analysis of Carthamus tinctorius L.
3.Investigation on psychological status and its relationship with Helicobacter pylori infection in servicemen from a Chinese army unit
Shujun LI ; Sa FANG ; Peng JIN ; Hui SU ; Hejuan AN ; Xiaowei WANG ; Xin WANG ; Jianqiu SHENG
Medical Journal of Chinese People's Liberation Army 2017;42(2):172-176
Objective To understand the mental health status of officers and soldiers (OSs) in a Chinese army unit,and analyze its relationship with Helicobacter pylori (H.pylori) infection.Methods Cross sectional extraction of 1293 male soldiers from a certain unit of Chinese army in September 2015,aged 17-37 years old,average age 20.5 ± 3.2.Using the symptom self rating scale (SCL-90) to conduct a psychological evaluation,comparison between the evaluation results and the Chinese military norm was conducted to understand the mental health status of OSs in the unit.13C-urea breath test was used to detect H.pylori infection,and the relationship between the positivity rate of various factors,the SCL-90 score and H.pylori infection were analyzed.Ninety OSs who had desire to eradicate H.pylori serves as the experimental group,and 90 H.pylori-infected OSs as control group.The experimental group received quadruple-drug eradication therapy,control group received no drug treatment.SCL-90 scores before and after the treatment were compared,and SCL-90 scores after treatment in the experimental group and those in the control group were also compared for analyzing the relationship between the OSs psychological status and H.pylori infection.Results The SCL-90 score of the OSs was lower than the norm of Chinese army,and the difference was statistically significant (P<0.05).The psychological screening positive rate was higher in H.pylori-positive OSs than H.pylori-negative those and the positive rates of psychological screening factors somatization,interpersonal sensitivity,hostility and paranoid were higher in H.pylori-positive OSs than in H.Pylorinegative those,and the difference was statistically significant (P<0.05).After the eradication of H.pylori in the experimental group these factor scores were lower than those before eradication.Compared with control group SCL-90 scores,soldiers somatization,interpersonal sensitivity,anxiety,hostility,paranoid factor score in experimental group after treatment were lower,the differences were statistically significant (P<0.05).Conclusion The mental health is better in OSs in this unit,the negative mental status is obvious in H.pylori-infected OSs than in H.pylori-negative those,and to eradicate H.pylori can improve their psychological state.
4.Amide proton transfer magnetic resonance imaging of Alzheimer's disease at 3.0 Tesla: a preliminary study.
Rui WANG ; Sa-Ying LI ; Min CHEN ; Jin-Yuan ZHOU ; Dan-Tao PENG ; Chen ZHANG ; Yong-Ming DAI
Chinese Medical Journal 2015;128(5):615-619
BACKGROUNDAmide proton transfer (APT) imaging has recently emerged as an important contrast mechanism for magnetic resonance imaging (MRI) in the field of molecular and cellular imaging. The aim of this study was to evaluate the feasibility of APT imaging to detect cerebral abnormality in patients with Alzheimer's disease (AD) at 3.0 Tesla.
METHODSTwenty AD patients (9 men and 11 women; age range, 67-83 years) and 20 age-matched normal controls (11 men and 9 women; age range, 63-82 years) underwent APT and traditional MRI examination on a 3.0 Tesla MRI system. The magnetic resonance ratio asymmetry (MTR asym ) values at 3.5 ppm of bilateral hippocampi (Hc), temporal white matter regions, occipital white matter regions, and cerebral peduncles were measured on oblique axial APT images. MTR asym (3.5 ppm) values of the cerebral structures between AD patients and control subjects were compared with independent samples t-test. Controlling for age, partial correlation analysis was used to investigate the associations between mini-mental state examination (MMSE) and the various MRI measures among AD patients.
RESULTSCompared with normal controls, MTR asym (3.5 ppm) values of bilateral Hc were significantly increased in AD patients (right 1.24% ± 0.21% vs. 0.83% ± 0.19%, left 1.18% ± 0.18% vs. 0.80%± 0.17%, t = 3.039, 3.328, P = 0.004, 0.002, respectively). MTR asym (3.5 ppm) values of bilateral Hc were significantly negatively correlated with MMSE (right r = -0.559, P = 0.013; left r = -0.461, P = 0.047).
CONCLUSIONSIncreased MTR asym (3.5 ppm) values of bilateral Hc in AD patients and its strong correlations with MMSE suggest that APT imaging could potentially provide imaging biomarkers for the noninvasive molecular diagnosis of AD.
Aged ; Alzheimer Disease ; diagnosis ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male
5.The electroporation effects of high power pulse microwave and electromagnetic pulse irradiation on the membranes of cardiomyocyte cells and the mechanism therein involved.
Hua DENG ; Dewen WANG ; Ruiyun PENG ; Shuiming WANG ; Jiankui CHEN ; Sa ZHANG ; Bo DONG ; Xiaomin WANG
Journal of Biomedical Engineering 2005;22(4):672-694
Though there is ongoing public concern on potential hazards and risk of electromagnetic radiation, the bioeffects mechanism of electromagnetic fields remains obscure. Heart is one of the organs susceptive to electromagnetic fields (EMF). This study was designed to assess the influence of high power pulse microwave and electromagnetic pulse irradiation on cardiomyocytes, to explore the critical mechanism of electromagnetic fields, and to explain the regular course of injury caused by exposure to pulse EMF. Cultured cardiomyocytes were irradiated by high power pulse microwave and electromagnetic pulse first, then a series of apparatus including atom force microscope, laser scanning confocal microscope and flow cytometer were used to examine the changes of cell membrane conformation, structure and function. After irradiation, the cardiomyocytes pulsated slower or stop, the cells conformation was abnormal, the cells viability declined, and the percentage of apoptosis and necrosis increased significantly (P< 0.01). The cell membrane had pores unequal in size, and lost its penetration character. The concentration of Na+, K+, Ca2+, Cl-, Mg2+, Ca2+ and P3+ in cell culture medium increased significantly (P< 0.01). and the concentration of Ca2+ in cells ([Ca2+]i) decreased significantly (P<0.01). The results indicated that cardiomyocytes are susceptible to non-ionizing radiation. Pulse electromagnetic field can induce cardiomyocytes electroporation, and can do great damage to cells conformation, structure and function. Electroporation is one of the most critical mechanisms to explain the athermal effects of electromagnetic radiation.
Animals
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Animals, Newborn
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Cell Membrane
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ultrastructure
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Cell Membrane Permeability
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radiation effects
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Cells, Cultured
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Electromagnetic Fields
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adverse effects
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Electroporation
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Mice
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Microwaves
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adverse effects
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Myocytes, Cardiac
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cytology
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radiation effects
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ultrastructure
6.Optimized strategy of anesthesia for thoracoscopic pulmonary surgery: erector spinae plane block with high volume of ropivacaine combined with general anesthesia
Guihuan CHEN ; Zixi GONG ; Qiuguo WANG ; Xin WANG ; Sa PENG ; Hui LI
Chinese Journal of Anesthesiology 2021;41(6):699-702
Objective:To evaluate the optimized efficacy of erector spinae plane block (ESPB) with high volume of ropivacaine combined with general anesthesia for patients undergoing thoracoscopic pulmonary surgery.Methods:Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of both sexes, aged 18-64 yr, with body mass index of 18-24 kg/m 2, scheduled for elective thoracoscopic pulmonary surgery, were divided into 2 groups ( n=30 each) using a random number table method: ESPB with high volume (40 ml) of 0.4% ropivacaine combined with general anesthesia group (E40 group) and ESPB with conventional volume (20 ml) of 0.4% ropivacaine combined with general anesthesia group (E20 group). Ultrasound-guided ESPB was performed on the operated side before general anesthesia induction in both groups.In E40 group, 0.4% ropivacaine 40 ml was injected.In E20 group, 0.4% ropivacaine 20 ml was injected.Total intravenous anesthesia was applied in both groups.Sufentanil was used for patient-controlled intravenous anesthesia (PCIA) after the end of operation.The PCIA pump was set up with a 2 ml bolus dose (0.04 μg/kg), a 10 min lockout interval and background infusion at a rate of 0.05 μg·kg -1·h -1.The analgesia lasted for 48 h after operation, and the numeric rating scale (NRS) score was maintained ≤ 3.When NRS score ≥ 4, morphine 5 mg was intravenously injected for rescue analgesia.The onset time of block, intraoperative consumption of propofol and remifentanil, time to first pressing the analgesia pump, the first requirement for morphine and the ratio of patients using morphine within 48 h after surgery were recorded.The cumulative consumption of requirement for opioids (effective pressing dose of analgesic pump+ consumption of requirement for morphine) within 12, 24, 36 and 48 h after surgery and satisfaction with analgesia were recorded.The extubation time, length of ICU stay, the first postoperative off-bed time, the chest tube removal time and duration of stay in hospital were recorded.The incidence of intraoperative adverse events (hypertension and hypotension) and postoperative adverse events (dizziness, nausea and vomiting) were recorded. Results:Compared with E20 group, the onset time of block was significantly shortened, the cumulative consumption of requirement for opioids within 24, 36 and 48 h after surgery was decreased, the ratio of paitents using morphine within 48 h after surgery was decreased, time to first pressing the analgesia pump and the first requirement for morphine were prolonged, satisfaction with analgesia score was increased, and the first postoperative off-bed time and the chest tube removal time were shortened in group E40 ( P<0.05). There was no significant difference in the incidence of adverse events between the 2 groups ( P>0.05). Conclusion:Compared with ESPB with the conventional volume of ropivacaine combined with general anesthesia, ESPB with the high volume of ropivacaine combined with general anesthesia can reduce the postoperative consumption of opioids, and prolong the duration of effective analgesia, which are helpful for postoperative rapid recovery without increasing the development of adverse events in patients undergoing thoracoscopic pulmonary surgery.
7.Comparison of diagnostic value between DR and MSCT in fracture and dislocation of foot and ankle.
Yong-ge LÜ ; Yong-liang TAN ; Jin-chao MO ; Rui-biao ZHENG ; Ding-kai YE ; Dong WU ; Di-lin LUO ; Sa PENG
China Journal of Orthopaedics and Traumatology 2013;26(7):553-556
OBJECTIVETo compare the diagnostic value between digital photography (DR) and multi-slice spiral CT (MSCT) in fracture and dislocation of foot and ankle.
METHODSFrom August 2010 to August 2012, the DR and MSCT data of 52 patients with fracture and dislocation of foot and ankle were compared according to results of surgery or discharge diagnosis. There were 37 males and 15 females, aged from 15 to 49 years old. Wilcoxon signed rank test was used for statistical analysis.
RESULTSThe results of 52 cases of MSCT were matched with the postoperative or discharge diagnosis. A total of 172 fractures were found on MSCT and 98 fractures were found on DR, the results had significant difference in detecting fracture (V=1 081, P<0.05); A total of 24 dislocations were found on MSCT and 16 dislocations were found on DR,the results also had significant difference in detecting dislocation (V=21, P<0.05). Fractures of 6 cases with DR diagnosis were corrected and located by MSCT.
CONCLUSIONMSCT is significantly better than DR in diagnosis of fracture and dislocation of foot and ankle. The examination of two parts should be performed in DR. MSCT and multi-planar reconstruction (MPR) examination should be further performed if DR results are unclear or do not match with clinical symptoms, missed diagnosis and misdiagnosis can be avoided.
Adolescent ; Adult ; Ankle Injuries ; diagnostic imaging ; Female ; Foot Injuries ; diagnostic imaging ; Fractures, Bone ; diagnostic imaging ; Humans ; Image Processing, Computer-Assisted ; methods ; Joint Dislocations ; diagnostic imaging ; Male ; Middle Aged ; Multidetector Computed Tomography ; methods ; Photography
8.Investigation on patient safety culture in standardized training for resident doctors
Qiong ZHAN ; Libin PENG ; La·dahesitan SA ; Yuan ZOU
Chinese Journal of Medical Education Research 2018;17(11):1171-1174
Objective To investigate the cognition on patient safety culture of resident doctors re-ceiving standardized training in two affiliated hospitals, analyze the effectiveness of publicity and education in patient safety culture and put forward suggestions for improvements. Methods A total of 913 resident doctors receiving standardized training in the two affiliated hospitals during 2014 to 2016 were enrolled. Their cognition on patient safety culture were investigated using questionnaire surveys from May to October in 2016, and the main factors influencing the cognition on patient safety culture in standardized training were put forward through expert interviews. All statistical analyses were performed with SPSS 17.0 software with Chi-square test. Results The response rate of the questionnaire was 87.62% (800/913). Three factors were involved in the patient safety system, including the hospital security objectives, security feedbackchannels, and adverse event warning mechanism. The cognitiive level of A hospital (68.96%, 62.52%, 62.81%) was higher than that of B hospital (52.99%, 46.16%, 47.01%), and the difference was statistically significant (P<0.05). The publicity of pre-service training, education in the department, and encouragement from teachers (82.13%, 84.48%, 78.33%) was better in A hospital than in B hospital (68.38%, 71.8%, 62.39%) (P<0.05). The reporting rate of adverse events in A hospital was higher than that in B hospital (P <0.05). Conclusion It is necessary for hospitals to further intensify the publicity and education on patient safety culture. We can improve students' cognition on patient safety culture to reduce medical errors by cre-ating a favorable atmosphere, establishing standardized admission education and training system, improving teachers' teaching ability, and strengthening the training for clinical skills.
9.Establishment of immune suppression-mediated infectious pneumonia model using resistant Acinetobacter baumannii strains
Zining TANG ; Xuewu LIU ; Qianqing LAN ; Hailiang TANG ; Sa XIAO ; Xiangzhong YUAN ; Dongdong PENG
Chinese Journal of Microbiology and Immunology 2023;43(12):908-917
Objective:To establish an animal model of pneumonia for research on clinical prevention and treatment of bacterial pneumonia by infecting immunocompromised rats with drug-resistant Acinetobacter baumannii ( Ab) strains. Methods:Drug-resistant Ab strains were selected. Forty-eight SD rats were randomly divided into four groups: normal group, cyclophosphamide control group (intraperitoneal injection of 45 mg/kg cyclophosphamide), bacterial infection group (intratracheal instillation of 1.5×10 8 CFU Ab suspension), and bacterial infection+ immunosuppression group (intraperitoneal injection of 45 mg/kg cyclophosphamide+ intratracheal instillation of 1.5×10 8 CFU Ab suspension). Flow cytometry analysis was used to detect the proportion of CD4 + , CD8 + and NK cells in rat peripheral blood before as well as 3 d and 7 d after infection. A lung function meter was used to detect peak inspiratory flow (PIF), peak expiratory flow (PEF), tidal volume (Vt ) and forced expiratory volume in the second second/forced vital capacity (FEV 200/FVC) at 3 d and 7 d after modeling. ELISA was used to detect the levels of IL-6, TNF-α and IL-10 in the alveolar lavage fluid. HE staining was used to observe the morphology of rat lung tissues in each group. Bacterial loads in rat lung tissues were counted by bacterial culturing. Results:A decrease in voluntary activity was observed in rats in the cyclophosphamide control group, bacterial infection group and bacterial infection+ immunosuppression group after modeling. Lung rales could be heard in the bacterial infection group and bacterial infection+ immunosuppression group. Compared with the normal group, the cyclophosphamide control group showed decreased proportion of CD4 + and CD11b + NK cells and increased CD8 + cells in peripheral blood; the bacterial infection group showed decreased PIF, PEF, Vt and FEV 200/FVC, increased IL-6 and TNF-α levels and decreased IL-10 level in the alveolar lavage fluid, and higher bacterial load in lung tissues with mild widening of alveolar walls and inflammatory cell infiltration ( P<0.05, P<0.01). Compared with the cyclophosphamide control group and the bacterial infection group, the bacterial infection+ immunosuppression group showed a lower proportion of CD4 + cells and a higher proportion of CD8 + cells in rat peripheral blood, decreased PIF, PEF, Vt and FEV 200/FVC, increased IL-6 and TNF-α levels and decreased IL-10 level in alveolar lavage fluid, higher bacterial load in lung tissues, and aggravated widening of alveolar walls and inflammatory cell infiltration ( P<0.05, P<0.01). The proportion of CD11b + NK cells in peripheral blood of rats in the bacterial infection+ immunosuppression group was significantly lower than that in the bacterial infection group ( P<0.05, P<0.01). Conclusions:A bacterial pneumonia model was successfully constructed by infecting rats with Ab alone or in combination with cyclophosphamide immunosuppression. In the model constructed with Ab and cyclophosphamide immunosuppression, the rats had more severe pneumonia, which might be related to the reduced cellular immune function and the aggravated bacterial infection in rat lung tissues by cyclophosphamide.
10.Construction and evaluation of an immunosuppression-mediated model of invasive Aspergillus niger lung disease in rats
Zining TANG ; Xiangchi CHEN ; Xuewu LIU ; Zhimin ZHOU ; Qiao LI ; Sa XIAO ; Dejian JIANG ; Dongdong PENG
Chinese Journal of Comparative Medicine 2024;34(6):63-72
Objective This study established a model of invasive Aspergillus niger lung disease in immunosuppressed rats to provide theoretical support for the pharmacodynamic evaluation of anti-invasive pulmonary aspergillosis drugs and mechanism studies.Methods Sixty SD rats were randomly divided into a normal control group;cyclophosphamide control group,and cyclophosphamide+fungal infection low,medium,and high dose groups,with 12 animals in each group.General clinical observations were performed daily,and the serum levels of immunoglobulin(Ig)G and IgM and galactomannan(GM)were detected by ELISA on the 3rd and 7th days of modeling.Simultaneously,the ratio of CD4+and CD8+cells,content of white blood cells(WBCs)and neutrophils(Neu)in peripheral blood,the Aspergillus niger load in alveolar lavage,and morphological changes to rat lung tissue were observed.Results Rats in the cyclophosphamide control and cyclophosphamide+fungal infection groups showed reduced voluntary activity and erect hair after modeling,and rats in the cyclophosphamide+fungal infection group also had shortness of breath and audible wet rhonchi in the lungs.Compared with the normal control group,rats in the cyclophosphamide control group showed significant reductions in the levels of CD4+,WBC,Neu,IgG,and IgM in the blood,and their proportion of CD8+cells was significantly higher(P<0.05,P<0.01).Compared with the cyclophosphamide control group,rats in the cyclophosphamide+fungal infection medium-and high-dose groups had significantly reduced blood levels of IgG,IgM,and CD4+cells(P<0.05,P<0.01);while the cyclophosphamide+fungal infection low-,medium-,and high-dose groups had significantly reduced blood levels of WBC and Neu(P<0.05,P<0.01).Additionally,rats in the cyclophosphamide+fungal infection medium-and high-dose groups had significantly increased blood CD8+cells(P<0.05,P<0.01),Blood GM levels and the alveolar lavage Aspergillus niger load were significantly increased in rats in the cyclophosphamide+fungal infection low-,medium-,and high-dose groups compared with the cyclophosphamide control group(P<0.05,P<0.01).The lung tissues of the cyclophosphamide+fungal infection low-,medium-,and high-dose groups showed mycelial distribution and destruction of alveolar epithelium,increase of bronchial epithelial cup cells in the alveoli,and infiltration of inflammatory cells,and the degree of lesions was positively correlated with the modeling dose.Conclusions In this study,we used Aspergillus niger combined with cyclophosphamide immunosuppressant to construct a model of invasive Aspergillus niger lung disease.The duration of the disease was positively correlated with the concentration of bacterial fluid and modeling time,confirming that cellular immunity plays an important role in the pathogenesis of the disease.At the same time,Ig can also affect the development of invasive pulmonary aspergillosis,and it is speculated that the pathogenesis may be related to the level of Ig produced by humoral immunity.