1.Application of genome editing technologies in researches on herpes simplex virus 1
Chinese Journal of Microbiology and Immunology 2016;36(6):472-476
The viral genome editing technologies have been shown to largely contribute to the stud-ies of viral gene functions and to the treatment of infectious diseases. With the development of genetic engi-neering technologies, a variety of viral genome mutation technologies have been established. The homologous recombination is traditionally used to edit the viral genome, but the low frequency of homologous recombina-tion limits its application. Large genome DNA viruses including herpes simplex virus 1 can be edited by bac-terial artificial chromosome system. However, the cloning of viral genome to bacterial artificial chromosome plasmid is both laborious and time-consuming, and parts of the plasmid genes or drug selection markers may remain in the genome of virus and then affect the function of virus. Fortunately, the CRISPR-cas9 system which was discovered in 2013 can be used to edit the viral genome efficiently and accurately. Furthermore, the experiment is simple and will not leave any trail of the viral genome when the genetic mutation is per-formed. Here, we briefly review the application of homologous recombination, bacterial artificial chromosome system and CRISPR-cas9 system in researches on herpes simplex virus 1.
2.Effect of hydrochloric acid and azithromycin in the treatment of children with mycoplasma pneumonia and its effect on the level of serum inflammatory factors in children
Xingli LEI ; Zhijian LAN ; Jun XU
Chinese Journal of Biochemical Pharmaceutics 2017;37(1):248-250
Objective To investigate the effect of the treatment of children with mycoplasma pneumonia and the level of inflammatory factors in the serum of children with mycoplasma pneumonia. Methods 180 cases of mycoplasma pneumonia in our hospital from July 2015 to June 2016 were randomly divided into observation group and control group,90 cases in each group. The control group were treated with azithromycin,and the observation group were treated with combination of hydrochloric acid and bromine on the basis of control group. The course of the two groups were all 12 days.The efficacy of the treatment, fever subsided time, the disappearance time of cough and pulmonary rales, before and after treatment serum inflammatory factor interleukin-6 (IL-6),interleukin-10 (IL-10),interleukin-17 (IL-17) and procalcitonin (PCT) levels between two groups were compared,and the incidence of adverse reactions. Results The observation group treatment efficiency 93.33% was higher than the control group 82.23% (P<0.05); The observation group dissipated time of fever,cough disappeared time and pulmonary rales disappeared faster than the control group (P<0.05);After treatment,the levels of serum IL-6, IL-17 and PCT in two groups were decreased,the level of IL-10 increased,the difference was statistically significant (P<0.05); the level of serum IL-6, IL-17 and PCT in observation group were lower than the control group, while the level of IL-10 was higher than that of the control group, the difference was statistically significant (P<0.05);There were no significant differences in the adverse reactions between the two groups . Conclusion The curative effect of bromhexine hydrochloride combined with azithromycin in treatment of mycoplasma pneumonia in children, and can reduce the serum inflammatory cytokines IL-6, IL-17 and PCT level and increase the level of IL-10 and lessen inflammatory reaction, has important research significance.
3.Matrix-assisted Laser Desorption/Ionizaiton-Mass Spectrometry Imaging of Oligosaccharides in Soybean and Leaf Using Ionic Liquid as Matrix
Xingli PEI ; Yuyu HUANG ; Can GONG ; Xu XU
Chinese Journal of Analytical Chemistry 2017;45(8):1155-1164
The ionic liquid butylamine 2,5-dihydroxybenzoate (DHB-BuN) was chosen as the matrix for improving repeatability of oligosaccharides quantitation, and imaging oligosaccharides in soybean and leaf by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS).Using the ionic liquid matrix methanol solution (20%, V/V) to cover sample slice, and setting MS acquisition parameters including polarity as positive and laser power as 70%, the MS imaging experiment was performed to analyze oligosaccharides and their distribution in soyben and soybean leaf.As a result, the MALDI-MS analysis of 3 kinds of oligosaccharides including sucrose, raffinose and stachyose was performed using the ionic liquid matrix and the reproducibility RSDs were less than 3% for in-spot and less than 4% for spot-to-spot.The standard curves linear correlation coefficients (R2) were all no less than 0.996 between MS peak intensity and oligosaccharides concentraions in the concentration rage of 0.062-1.00 mg/mL, which showed quantitative power in MALDI-MS analysis.The matrix was used for MALDI-MS imaging analysis of soybean chips and soybean leaf surface disaccharide, trisaccharide and tetrasaccharide to obtain the soybean mass spectrum and spatial resolution of 150 μm of the three oligosaccharides imaging.Three kinds of oligosaccharides in the soybean roughly balanced distribution, and the oligosaccharides distribution in the leave was mostly at its tip part.The contents of the 3 kinds of oligosaccharides could be estimated by their MS peak intensity and above standard curve.The result suggested that the ionic liquid DHB-BuN matrix could be well applied in MALDI-MS imaging analysis of oligosaccharides in different sample.
4.Surgical strategy for treatment of type A aortic dissection with small true lumen of the descending aorta
Yangfeng TANG ; Lin HAN ; Xingli FAN ; Boyao ZHANG ; Jiajun ZHANG ; Qing XUE ; Jie LU ; Zhiyun XU
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(4):208-211
Objective:To summarize the results and methods of surgical treatment for type A aortic dissection with small true lumen of the descending aorta.Methods:9 patients underwent surgical treatment for type A aortic dissection with small true lumen of the descending aorta between January 2017 and December 2019 were analyzed retrospectively. There were 7 males and 2 females, mean age of (41.6±9.2) years. Acute dissection were 2 cases, and chronic dissection were 7 cases. Preoerative computed tomography was used to diagnose the dissection and evaluate the true lumen of the descending aorta. This procedure was done in all patients via a median sternotomy under hypothermic CPB with SCP. 4-branched prosthetic graft was used to replace the ascending aorta and aortic arch. The procedures involving the descending aorta: Hybrid surgery using TEVAR. Distal intimal flap fenestration. Implanting the intraoperative stent-graft or prosthetic graft at false lumen for second-step operation.Results:There was no in-hospital mortality. Stroke, Spinal cord, visceral ischemia and lower limbs malfunction were not observed. Reintervention was not found in case with acute dissection during follow-up. One patient who reveived fenestration underwent TEVAR, others with chronic dissection underwent thoracoabdominal aortic replacement 3 months after surgery.Conclusion:Hybrid or staged procedures was a suitable alternative to patients with type A aortic dissection with small true lumen of the descending aorta.
5.Cloning of UL7 protein of HSV-1 and analysis of the expression of UL7 protein during the prolifera-tion of HSV-1
Jienan ZHOU ; Xingli XU ; You GAO ; Ying ZHANG ; Jingjing WANG ; Longding LIU ; Qihan LI
Chinese Journal of Microbiology and Immunology 2016;36(4):241-246
Objective To express and purify the recombinant UL7 protein of herpes simplex virus 1 (HSV-1), to prepare the corresponding UL7-specific polyclonal antibody and to preliminarily analyze the expression of UL7 protein during the proliferation of HSV-1. Methods The UL7 gene was amplified by PCR and then cloned into the pGEX-5X-1 vector for expression of UL7 protein in the prokaryotic expression system. The constructed expression plasmid, pGEX-5X-1-UL7, was transformed into E. coli BL21 (DE3) to induce the expression of UL7 protein by IPTG. The purified GST-UL7 fusion protein was used as antigen to inject the ICR mouse for the preparation of polyclonal antibody specific for UL7 protein. The titer and speci-ficity of the polyclonal antibody were analyzed by using indirect ELISA and Western blot assay, respectively. The UL7 protein-specific polyclonal antibody was used to detect the expression of UL7 protein at different time points after infecting Vero cells with HSV-1. Results The GST-UL7 fusion protein was efficiently ex-pressed in E. coli BL21 (DE3). The UL7 protein-specific polyclonal antibody was prepared with high titer (1 ∶ 105) and high specificity as indicated by the indirect ELISA and Western blot assay. The expression of UL7 protein was detected at different time points after infecting Vero cells with HSV-1. Conclusion The GST-UL7 fusion protein was obtained successfully and the UL7 protein-specific polyclonal antibody was pre-pared. Accompany with the proliferation of HSV-1, the expression of UL7 protein was detected at different time points by using the polyclonal antibody.
6.Serum level and clinical significance of macrophage inflammatory protein-1α and interferon ;gamma inducible protein-10 in acute myelogenous leukemia
Xingli ZHANG ; Pengfei WU ; Rong KONG ; Hongchun QIU ; Dehong WU ; Jing XU
Chinese Journal of Postgraduates of Medicine 2017;40(2):171-174
Objective To study the serum level of macrophage inflammatory protein-1α(MIP-1α) and interferon gamma inducible protein-10 (IP-10) in acute myelogenous leukemia (AML) and clarify their clinical significance. Methods Enzyme-linked immunosorbent assay was used to detect the level of MIP-1α and IP-10 in serum samples from 34 AML patients(observation group) and 20 volunteers (normal control group). Results The levels of MIP-1αand IP-10 in observation group before induction chemotherapy were significantly higher than those in normal control group (P<0.05). The levels of MIP-1αand IP-10 in observation group after induction chemotherapy were decreased, and significantly lower than those before induction chemotherapy (P<0.05). After treatment for one course, 21 patients reached complete remission (CR), and 13 patients did not reach CR. The levels of MIP-1αand IP-10 in CR group had no significant difference compared with those in normal control group (P<0.05), but the levels of MIP-1αand IP-10 in none CR group were significantly higher than those in normal control group and CR group (P<0.05). The drop percentage of MIP- 1αlevels in CR group and none CR group was (32.51 ± 10.34)% and (10.57 ± 10.39)%, and there was significant difference (P<0.05). The drop percentage of IP-10 levelsin CR group and none CR group was(45.94 ± 13.68)% and (31.17 ± 11.85)%, and there was significant difference (P<0.05). Liner correlation analysis revealed that the levels of MIP-1αand IP-10 had significantly positive correlation in AML patients (r=0.652, P<0.05). Conclusions Different expressions of serum MIP-1α and IP-10 are found before and after induction chemotherapy AML patients, and there is significant correlation. Combined detection of serum MIP-1αand IP-10 may be used as an index to monitor clinical stages and prognosis.
7.A Meta-analysis of the incidence of influenza reported during year 2005 to 2012 in China
Yunguang HU ; Xingli XU ; Jingjing WANG ; Jie SONG ; Yancui WANG ; Haiwei LI ; Longding LIU ; Haijing SHI
Chinese Journal of Microbiology and Immunology 2015;(4):265-270
Objective To investigate the epidemic patterns and the characteristics of influenza in Chi-na through a Meta-analysis based on the studies published in domestic literatures.Methods Related articles published during 2005 to 2012 were screened out from domestic databases and analyzed through a Meta-analysis with Review Manager 5.0 software.Results Twenty-two articles covering 957 901 patients with influenza-like-illness (ILI) and 148 233 pathogen samples were screened out according to the inclusion criteria.No significant difference with the ILI diagnosis rate was found between subjects at age 0-4 years and those at age 15-59 years. Higher ILI diagnosis rates were observed in those two groups as compared with subjects elder than 60 years old. Most of the pathogen samples were carried by subjects aged 25-59 years.More influenza virus strains were isola-ted in 2009 as compared with those of the seven other years (OR=2.25, 95%CI=1.27-3.70).There was sta-tistical difference between the numbers of influenza A H1N1 and seasonal influenza A strains (OR=2.25, 95%CI=1.30-3.91) .Significant difference was also observed between the numbers of influenza A and influenza B strains (OR=4.05, 95%CI=2.53-6.47).Conclusion There was significant difference with the diagnosis rate between subjects aged 0-4 years and those aged≥60 years.More attention should be paid to people at high risk of infection (0-4 years old and≥60 years old) and those at 25-29 years with high mobility and social inter-course for the timely prevention and control of pandemic influenza.The detection rate of influenza virus strains was increased during the outbreak of novel influenza A H1N1 infection in 2009.After that outbreak, the detec-tion rate of novel influenza A H1N1 strains was 2.25 times the rate of seasonal influenza strains.The detection rate of influenza A was 4.05 times the rate of influenza B virus strains.Therefore, it is necessary to strengthen the surveillance for influenza A virus and other epidemic influenza virus strains.
8.Effects of IGF-1 and oxLDL on expression of phosphatase PHLPP1 in vascular smooth muscular cells
Xingli WU ; Dingyou YANG ; Zhongsu YANG ; Deyin LI ; Huibin XU ; Shiwen WANG
Journal of Geriatric Cardiology 2009;6(4):237-240
Objective To investigate the effects of insulin-like growth factor-1 (IGF-1) and oxidized low density lipoprotein (oxLDL) on expression of phosphatase PHLPP1 in vascular smooth muscle cells (VSMCs).Methods Rabbit aortic VSMCs were cultured.VSMCs proliferation ability was determined by measuring cell number and mitochondrial dehydrogenase (MD) activity with MTT assay.Western blot was used to detect the protein expression of phosphatase PHLPP 1.Results IGF-1 (100μg/L) increased cell number and MD activity to 3.02 and 3.59 times of that in control group.oxLDL(50μg/ ml) elevated the above two parameters to 2.03 and 2.91 times respectively.Western blot showed that IGF-1 and oxLDL inhibited the expression of PHLPP 1 to 39.27% and 40.26% of the control group (P<0.01).Conclusion IGF-1 and oxLDL may enhance the proliferation of VSMCs by decreasing the expression of phosphatase PHLPP1 .
9.The predictive value of clinical and radiographic features in fungal pathogen identification in immunocompromised patients with pulmonary invasive fungal infection
Xingli GU ; Mingqin CAO ; Sicheng XU ; Qiufeng WAN ; Guangming LIU ; Jing WANG ; Wenya LIU
Chinese Journal of Emergency Medicine 2016;25(7):920-926
Objective To investigate the predictive value of clinical and radiographic features in fungal pathogen identification in immunocompromised patients with pulmonary invasive fungal infection (IFI).Methods All consecutive immunocompromised adult patients with pulmonary IFI in respiratory intensive care unit (ICU)in the First Affiliated Hospital of Xinjiang Medical University were recruited during a 2 year period.All patients met the 2008 European Organization for Research and Treatment of Cancer and Mycoses Study Group (EORTC /MSG) criteria were studied for proved or probable IFI responding to antifungal agents.The data of demographic,clinical and radiographic features,as well as serological test results of the patients were collected.Differences in the clinical and radiographic features of pulmonary IFIs caused by yeasts and molds were compared by χ2 test.A logistic regression model was used to perform discriminant analysis,and the effect of discrimination was assessed for accuracy.Results The study included 143 patients with a probable diagnosis of IFI who had the following risk factors:diabetes mellitus (43.4%),chronic lung disease (32.2%),broad-spectrum antibiotics administration (≥14 days;35.7%),malignancy (23.1%),corticosteroid therapy (≥14 days;23.1%),chronic renal failure and renal replacement therapy (16.1%),and immunological disease (10.5%).Frequent broad-spectrum antibiotics administration was associated with yeast infection (P <0.05 ),while mold infection was associated with chronic lung disease (P <0.05 ) .Yeast was more often isolated from patients with concurrent bacterial infection and on mechanical ventilation (P <0.05 ) . Thoracic high-resolution computed tomography (HRCT)showed the following images:bronchial pneumonia/pulmonary consolidation (53.1%),massive shadowing (29.4%),small nodules (24.5%),large nodules (18.9%),pleural effusion (18.9%),halo sign (14%),and cavity (9.8%).Imaging showed that mold was more common than yeast in patients with pleural and pericardial effusions (P <0.05).Logistic regression modeling showed that broad-spectrum antibiotics administration,prolonged mechanical ventilation,and pleural and pericardial effusions were statistically significant in fungal identification (P <0.05 ),with a predictive accuracy of 77.6%.Conclusions For immunocompromised patients with pulmonary IFI,most of the risk factors ,the main clinical and chest HRCT features did not help to predict the type of fungal pathogen,and yeast but not cryptococcus may be accompanied or colonized.
10.Exploration on Permanent Staining Preparation of Tapeworm
Shengxia CHEN ; Liang WU ; Huijuan XU ; Lianyun SHUAI ; Xingli FU ; Jinbo QIU
Chinese Journal of Parasitology and Parasitic Diseases 2006;0(S1):-
Objective To make staining preparation of tapeworm for the purpose of teaching and scientific research.Methoeds Pregnant segmens were stained by Indian ink,and mature segments,cephalomeres and bladder worms were stained by carmine.Result After staining the characteristics of pregnant segments,mature segments,cephalomeres and bladder worms were observed obviously.For instance,the stem and branches linking both-side of uteri in pregnant segment and reproduction cavity projecting in one of edges in the center of the pregnant segment were clearly shown in ink colour after stained by Indian ink.The red colour was shown in the mature segment with deep-coloured male and female reproduction systems inside,separately,after stained by carmine.Conclusion Tapeworm preparation stained by Indian ink and carmine is useful and can be preserved permanently.