1.Differences of cellular apoptosis in lungs between C57BL/6J and C3H/HeN mice in the course of radiation induced pulmonary injury
Xiao ZHANG ; Yang LI ; Lei-Lei YANG ; Shuo TANG
Military Medical Sciences 2017;41(10):787-791
Objective To explore the difference of cellular apoptosis levels in lungs between C 57BL/6J and C3H/HeN mice in the course of radiation induced pulmonary injury (RPI).Methods Sixty C57BL/6J mice and another sixty C3H/HeN mice were randomly divided into control group and irradiation group respectively .The 60 Co γray irradiation method was used to establish an RPI model with 20 Gy per irradiation.Lungs were obtained on 1 d, 3 d, 7 d, 1 month(m) and 3 m after irradiation, and then the apoptosis and expressions of γH2AX and AIF were detected by in situ terminal labeling and immunoblotting .Results 1 and 3 d after irradiation , many apoptotic cells , mainly apoptotic epithelial cells , could be seen in lungs and apoptotic cells in the lungs of C 57BL/6J mice outnumbered those of C3H/HeN mice.At one month after irradiation, there was no significant difference of cell apoptosis between these two groups of mice .Three months after irradiation, there were fewer apoptotic cells in lungs of C 57BL/6J mice than in C3H/HeN mice.C57BL/6J mice had a higher γH2AX expression in lungs than C3H/HeN mice did at 1 -7 days after irradiation.However, the expression ofγH2AX in lungs of C3H/HeN mice was higher than that of C57BL/6J mice at 3 m after irradiation.The expression of AIF in lungs of C3H/HeN mice was lower than that of C57BL/6J mice at 1 d, 3 d and 1 m,while the expression of AIF in lungs of C3H/HeN mice was higher than that of C57BL/6J mice at 3m after irradiation.Conclusion In the course of RPI, the occurence of DNA damage and cell apoptosis is different in lungs of C 57BL/6J mice and C3H/HeN mice.
2.Construction of pEGFP-C1-CXCL1 plasmid and detection of its expression in eukaryotic cells
Rui-Tao MAO ; Wei CHEN ; Zi-Hui LI ; Ling ZOU ; Jia-Zhou YE ; Tao BAI ; Jie CHEN ; Jian-Kang CHEN ; Cui WANG ; Ning LIU ; Xiao-Li YANG ; Cong-Wen WEI ; Hui ZHONG ; Fei-Xiang WU
Military Medical Sciences 2017;41(10):792-795
Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.
3.The role of miR-155/SOCS1 pathway in collaboration with IL-17 in modulating the early progression of Schistosoma japonica
Tan LI ; Wei-Jie YOU ; Xin GE ; Ping MA ; Jie XING
Military Medical Sciences 2017;41(10):796-799
Objective To explore the role of microRNA-155 ( miR-155 )/suppressor of cytokine signaling 1 ( SOCS1 ) pathway combined with IL-17 in modulating the early progression of Schistosoma japonicum.Methods BALB/c mice were infected subcutaneously with cercariae of S.japonicum before being sacrificed at 8 and 14 weeks after infection to collect liver samples for pathological observation by H&E staining .The spleens of mice were collected to calculate the spleen index.The level of IL-17 in serum was determined using ELISA.Furthermore, qRT-PCR was used to detect the expression of miR-155 and SOCS1 mRNA in the spleens of mice .Results Compared with normal group , the spleen index in the model group was increased significantly along with the loss of relative body mass .H&E staining revealed that the deposition of parasite eggs began to appear in the liver tissue at week 8 post-infection.Meanwhile, there were a large number of inflammatory cells infiltrating to form the eosinophilic granuloma.Over time, the formation of egg granulomas and the infiltration of inflammatory cells were alleviated on week 14 post-infection.The level of IL-17 in serum and the expression of miR-155 in the spleen were peaked on week 8 post-infection, and decreased sharply on week 14, but were still evidently higher than those in normal group .The expression of SOCS1 mRNA was progressively increased both on week 8 and 14. Conclusion The data indicates that miR-155 might cooperate with IL-17 to modulate SOCS1 in the development of S.japonicum infection.
4.Construction of sRNA-deficient and-overexpressing strains of Yersinia pestis
Xiao-Fang GAO ; Zi-Zhong LIU ; Wen-Liang LI ; Hong-Duo WANG ; Rui-Fu YANG ; Yan-Ping HAN
Military Medical Sciences 2017;41(10):800-804
Objective To construct small RNA deletion and overexpression strains with a length of less than 100 nt in Yersinia pestis.Methods Deletion mutants of the target sRNAs were constructed by increasing the length of homologous regions.Meanwhile, the high copy plasmid pBAD/HisA was modified into an inducible transcriptional vector as an sRNA-overexpression plasmid by using QuikChange lightning site-directed mutagenesis kit .The presence , size, and transcription-al initiation sites of the indicated sRNA were predicted by transcriptome sequencing , primer extension , and previous stud-ies.The full-length DNA fragments of target sRNAs were transformed into the transcriptional vector .The overexpressing strains of sRNAs were identified by Northern Blot .Results and Conclusion Four sRNAs deletion mutants of sR01, sR02, sR03 and HmsA and three sRNAs overexpression mutants MicF , HmsA and CpxQ were successfully constructed .A method of construction of sRNA deficient and overexpressing strains of Y.pestis has been quickly and efficiently established by λ-Red homologous recombination technology and QuikChange ? lightning site-directed mutagenesis kit.
5.Biological characteristics and biofilm formation of Klebsiella pneumoniae TH6 strain
Bo-Xing LI ; Jing LU ; Wei-Wei CHENG ; Zhi-Xi LI ; Jing YUAN
Military Medical Sciences 2017;41(10):805-809
Objective To explore the biological characteristics and biofilm formation of a strain of Klebsiella pneumoniae isolated from the stool of a patient with clinical inflammatory bowel disease .Methods The TH6 strain was identified as Klebsiella pneumoniae by 16S RNA, MALDI-TOF and automatic biochemical instrument .The morphology, growth rate, biofilm formation, capsid virulence factors and other biological characteristics were analyzed .Results and Conclusion The TH6 strain was short rod-shaped with a capsule under the transmission electron microscope , and was of K57 type according to the capsule serotype test .K.pneumonia TH6 grew rapidly under aerobic and anaerobic conditions , almost at twice the rate of standard strain ATCC2146, and reached the logarithmic growth phase at 2 h.The strain had multi-drug resistance.Biofilm formation was observed under a confocal laser scanning microscope (CLSM) at 4, 8, 12, 24, 48, 72, 96 and 120 h and the biofilm was formed at 72 h.The ability of fast growth , biofilm formation and other characteristics of K.pneumonia TH6 showed that this strain has a strong ability to adapt to the new environment is difficult to cure after colonization in vivo.
6.Preformulation study on novel antiviral drug GZ914
Guan-Hua HOU ; Xiao-Yang HAN ; Yan-Jun CHEN ; Qian JIN ; Yu-Li WANG ; Qiang WU
Military Medical Sciences 2017;41(10):810-813,863
Objective To study the physical-chemical parameters of antiviral drug GZ 914 and provide data for the preparation design .Methods The appearance , crystal structure and solubility of GZ 914 were investigated .An HPLC method was established to determine the content of GZ 914 in vitro before oil/water partition coefficient and solubility in different pH experiments were calculated .Results GZ914 was a straw yellow powder with a crystalline structure , low water-solubility and good lipotropy .The HPLC method had a good linear relationship within the range of 12-60 μg/ml (r=0.9998).The oil/water partition coefficient of GZ914 was 1.9.Conclusion This analytical method is accurate and reliable.The oil/water partition coefficient indicates that the drug could be formulated as an oral solid preparation.
7.Epidemiological characteristics of human adenovirus associated respiratory diseases in China
Wei CHEN ; Sheng-Shu WANG ; Wen-Yi ZHANG ; Quan QIAN ; Liang WEN ; Yuan-Yong XU ; Qing-Hua LI ; Hai-Long SUN ; Sheng-Long LI ; Yong WANG
Military Medical Sciences 2017;41(10):814-821
Objective To analyze the epidemiology of outbreaks and epidemic characteristics of respiratory diseases caused by human adenovirus in China so as to provide some data for its epidemic and outbreak control and clinical diagnosis .Methods Data on respiratory adenovirus outbreaks and surveillance from 1997 to 2015 was collected from PubMed, China National Knowledge Infrastructure (CNKI), and Wanfang Databases.All the data was analyzed according to the descriptive epidemiology , including the time , area and population distribution .Clinical data and the serotypes of adenovirus were also analyzed.Results From 1997 to 2015, the epidemical serotypes of adenovirus included 1 to 7, 11, 14 and 55 in China, and the dominating serotypes were 7 and 3, which accounted for 62.33%(599/961) and 24.97%(240/961)of the total cases of outbreaks, and for 36.79%(312/848) and 53.18%(451/848) of the total cases of surveillance.The peaks of annual outbreaks were in 2004 and 2013, which made up 41.12%(2212/5380) and 16.49%(887/5380)of the total outbreak cases in this study .Most of the surveillance cases years occurred in 2010 and 2011, which accounted for 17.59%(297/1688) and 17.77%(300/1688) of the total cases of surveillance .The seasonal distribution of the outbreaks was characterized by the highest possibility in spring and winter .Outbreaks of respiratory adenovirus were reported by 12 provinces or municipalities .The number of reported outbreaks related to serotype 3 was the largest in Jiangsu Province, which made up 58.33%(140/240) of the total.Most of the reported cases related to serotype 7 occurred in Hubei Province, which made up 67.41% (333/494) of the total.Most of cases were found in Peking and Jiangsu , which accounted for 57.56%(971/1687)and 32.42%(547/1687)of the total positive cases respectively.The high-risk populations were children and new recruits , who accounted for 73.97%(2907/3930) of the total.The clinical features of adenovirus infection were fever (63%-100%),sore throat (31.9%-100%), pharyngeal hyperemia (60%-100%) and cough (5.88% -100%).Conclusion Human respiratory adenovirus has become one of the main pathogenic microorganisms that induce acute respiratory diseases in schools and in the military in China , so human adenovirus and related respiratory disease should be monitored in such populations .The epidemiological characteristics of different types of respiratory adenovirus and the patterns of spread should be analyzed in order to reduce morbidity and mortality.
8.Serological characteristics and gene mutation analysis of Para-Bombay blood group
Xiao-Qin ZHOU ; Zhi-Hui SHEN ; Nai-Cong ZHANG ; Song JIN ; Sheng-Qiang LIANG
Military Medical Sciences 2017;41(10):822-824
Objective To identify the Para-Bombay blood group on the basis of its serological characteristics .Methods ABO blood typing , H antigen detection , absorption and elution test , and saliva neutralization test were conducted for serological identification of ABO blood group .PCR-SSP was used to sequence FUT1 and FUT2 genes.Results Results of ABO genotyping of eight individuals of the Para-Bombay blood group were consistent with results of their serological blood typing.Among these cases, there were 3 cases of Amh,4 cases of Bmh,and 1 case of Abmh.The results of their FUT1 genotyping were h1h1 in 3 cases, h2h2 in 2 cases and h1h2 in 3 cases.Conclusion The differentce of agglutination intensity between Ac and Bc in reverse ABO blood typing and abnormal Oc agglutination is of greet significance for Para -Bombay blood group.
9.Diagnostic efficiency and result of prostate biopsy for patients with t-PSA between 4.0 and 10 ng/ml
Jian-Nan LIU ; Hai-Xing MAI ; Nan QU ; Xue-Chao LI ; Ya-Lin WANG ; Chen HUANG ; Li-Jun CHEN
Military Medical Sciences 2017;41(10):825-829
Objective To investigate the diagnostic efficiency and result of prostate biopsy for patients with t -PSA between 4.0 and 10 ng/ml.Methods This analysis was based on 20 qualified research papers from such lectronic databases as PubMed, MEDLINE, EMBASE and Cochrane from January 2010 until September 2017.Data extracted was analyzed using classic Meta-analysis with R software .The random or fixed effect model analysis was used to estimate the rate.Heterogeneity was analyzed using I 2 statistic.Results Totally 5481 patients were included in the 20 research papers. The positive rate of prostate biopsy was 20.6%, with t-PSA between 4.0 and 10 ng/ml, which was higher than the rate in the data from CUA Guide(2014).The difference was statistically significant .Conclusion Patients should be subjected to prostate biopsy if their t-PSA ranges 4.0 from 10 ng/ml regardless of the rate of f/t-PSA.The Gleason grade is relatively low when PSA is in the gray area , and the risk is also low.
10.Clinical study of decitabine-intensified conditioning regimen prior to hematopoieticstem cell transplantation in the treatment of refractory/relapse acute myeloid leukemia
Long-Can CHENG ; Na LIU ; Jian-Lin CHEN ; Bo-Tao LI ; Chen XU ; Yu-Hang LI ; Liang-Ding HU
Military Medical Sciences 2017;41(10):830-834
Objective To observe the treatment results of 44 consecutive patients with refractory/relapse acute myeloid leukemia( AML) not in remission who received allogeneic hematopoietic stem cell transplantation ( allo-HSCT ) following decitabine (DAC)-intensified conditioning regimen (18) and idarubicin(IDA)-intensified conditioning regimen (26). Methods We conducted a retrospective study to evaluate the outcome of 44 consecutive patients with refractory/relapse AML not in remission who received allo-HSCT from 2009 July to 2016 May.Eighteen of them were given DAC-intensified conditioning regimen and 26 of them were given IDA-intensified conditioning regimen prior to allo-HSCT.The effects of DAC and IDA intensified conditioning regimen on the patients ' engraftment, transplant-related mortality, survival and occurrence of graft versus host disease(GVHD)were analyzed.Results and Conclusion In the DAC group, 7(38.9%) patients had acute GVHD (aGVHD).Among them,2 patients had grade Ⅰ aGVHD,5 patients had grade Ⅱ aGVHD.In the IDA group, 16(61.5%)patients had aGVHD.Among them,9 patients had grade Ⅰ aGVHD, 6 patients had grade Ⅱ aGVHD and 1 patient had grade ⅢaGVHD.In the IDA group, 9 of 26(34.6%) patients experienced leukemia relapse , all of them died due to the lack of effective therapies .In the DAC group, 4 of 18(22.2%) patients experienced leukemia relapse, 2 of them got long-term survival throughout salvage therapies .For the DAC group and IDA group , the 1-year probabilities of overall survival (OS) and leukemia-free survival (LFS) were 65.0% versus 57.7% (P=0.602) and 53.5%versus 57.7%(P=0.910), respectively.DAC-intensified conditioning regimen before allo-HSCT in the treatment of refractory/relapse AML is safe and effective.