2.Molecular characteristics and RNA interference efficiency of Schistosoma ja-ponicum Sj79 gene
Yanyan JIANG ; Zhongying YUAN ; Yuxin XU ; Wei ZANG ; Jianping CAO ; Ying WANG ; Jianhai YIN ; Yujuan SHEN
Chinese Journal of Schistosomiasis Control 2015;(3):273-276,289
Objective To study the structural features and characteristics of a novel gene Schistosoma japonicum 79(Sj79), and observe its effect of RNA interference(RNAi),so as to provide the experimental basis for its further function study and mechanism study of anti reproductive development of schistosome. Methods The gene structure and characteristics of Sj79 were analyzed by bioinformatics methods. Then the expressions of Sj79 messenger RNA(mRNA)during the different develop?mental stages of schistosome were analyzed and the effects of RNAi silencing were observed by the soaking method. The tran?scriptional levels of Sj79 after RNAi were detected by real time PCR. Results The open reading frame of Sj79 contained 696 base pairs with an exon structure. The gene had obvious stage specificity,and its transcriptional level in mature female worms was the highest. After soaking for 3 d,the Sj79 mRNA level[(41.0 ± 12.3)%]in the siRNA?1 group with low dosage(20 nmol/L) was lower than that in the siRNA?NC group[(103.2 ± 14.4)%],the difference was statistically significant(t=3.28,P<0.05). When with high dosage(200 nmol/L ),both the Sj79 mRNA levels in the siRNA?1 group[(15.8 ± 10.9)%]and siRNA?2 group [(11.1 ± 8.8)%]were significantly lower than that in the siRNA?NC group[(100.1 ± 6.3)%](t=13.44,27.84,both P<0.01). After soaking for 7 d,only the Sj79 mRNA levels in the siRNA?1group[(43.4 ± 4.5)%]and siRNA?2 group[(62.5 ± 5.4)%]with low dosage were lower than that in the siRNA?NC group[(100.4 ± 5.2)%],and the differences had statistical sig?nificance(t=8.33,5.07,both P<0.01). Conclusion Through this study,we have improved the mRNA sequence and genom?ic information of Sj79 gene,and understood its structural features,as well as selected out two effect fragments siRNA?1and siR? NA?2 which will provide the basic evidences for the further study on egg laying interference of the female adult worm of schisto?some in vitro.
3.Immunoscreeening of Schistosoma japonicum schistosomula cDNA library with sera from Microtus fortis and bioinformatic analysis of novel genes
Zhongying YUAN ; Yujuan SHEN ; Jianping CAO ; Jijia SHEN ; Yuxin XU ; Wei DIAO ; Yuan HU ; Xiaohong LI ; Shuxian LIU
Chinese Journal of Schistosomiasis Control 1989;0(04):-
Objective To obtain novel vaccine candidate antigens against Schistosoma japonicum. Methods S. japonicum schistosomula cDNA library was screened by using sera of Microtus fortis that was naturally resistant to schistosomiasis. The positive clones were transformated into Escherichia coli BM25.8, E. coli clones containing the plasmid cultured in LB, and then selected for plasmid extraction, the plasmid DNA was digested by EcoRⅠand Hind Ⅲ, and analysed by agarose gel electrophoresis. The positive clones were also sequenced and the data were analysed through the internet Nucleotide BLAST software of NCBI and Expert Protein Analysis system of GeneRunner and HNN. Results Twelve positive clones were obtained after repeatedly immunoscreening the library and their sizes ranged from 300 bp to 1100 bp. Two novel genes (named as Sj-sMf1 and Sj-sMf2) with complete ORF were obtained. The deduced protein of Sj-sMf1 consisted of 93 amino acids while Sj-sMf2 consisted of 61 amino acids. Sj-sMf1 protein predicted containing one cAMP phosphorylation site and Casein kinase C phosphorylation site, respectively. Sj-sMf1 protein predicted containing one Casein kinase C phosphorylation site and two Protein kinase C phosphorylation sites. Conclusion Two novel genes predictably encoding unknown proteins are obtained from immunoscreening of Schistosoma japonicum schistosomula cDNA library by M. fortis sera.
4. Clinical observation of continuous and intermittent application of lamivudine or entecavir resistance mutations in patients with chronic hepatitis B
Shuhong DUAN ; Xiaodong YUAN ; Xiaoyan LIU ; Lei WANG ; Rui BAI ; Ting PANG ; Zhongying BAO
Chinese Journal of Hepatology 2018;26(9):646-649
Objective:
To observe continuous and intermittent application of lamivudine or entecavir resistance mutations in patients with chronic hepatitis B.
Methods:
Data of patients with active stage of chronic hepatitis B over the past 6 years were collected and analyzed retrospectively. The incidence of drug resistance mutation and related factors between patients taking LAM or ETV continuously and intermittently were compared with those taking LAM or ETV. Data comparison was performed using
5.Relationship between pre-hospital delay and health belief in patients with colorectal cancer
Xiaodan WU ; Na LI ; Lifang YUAN ; Huiying QIN ; Lijuan ZHANG ; Zhongying HUANG ; Meichun ZHENG
Chinese Journal of Practical Nursing 2017;33(30):2361-2365
Objective To describe the current situation of pre-hospital delay and health belief in patients with colorectal cancer and explore the relationship between them. Methods With convenient sampling methods,totally 385 patients with colorectal cancer were recruited from January to June 2016. Patients with colorectal cancer were investigated using a general data survey questionnaire, visit situation questionnaire and the Champion Health Belief Model Scale. Results The median pre-hospital delay time was 3.00 months.The rate of pre-hospital delay was 43.1%(166/385). The score of the overall health belief was (119.52 ± 12.62) points. The two dimensions of"perceived susceptibility and perceived severity" scored lower, and the two dimensions of "health motivation and perceived therapeutic benefits" scored higher. The health belief was negatively correlated with the pre- hospital delay among colorectal cancer patients (r=- 0.737, P<0.01). Conclusions Pre-hospital delay occurred among patients with colorectal cancer is severe. Patients with a higher level of health belief have shorter pre-hospital delay.It′s important to improve the level of health belief of"perceived susceptibility and perceived severity"and reduce the time of pre-hospital delay to improve outcomes in patients with colorectal cancer.
6.The identification of the Cryptosporidium ubiquitum in pre-weaned Ovines from Aba Tibetan and Qiang autonomous prefecture in China.
Yujuan SHEN ; Jianhai YIN ; Zhongying YUAN ; Weiyuan LU ; Yuxin XU ; Lihua XIAO ; Jianping CAO
Biomedical and Environmental Sciences 2011;24(3):315-320
OBJECTIVECryptosporidium spp. are prevalent globally and sheep are an important zoonotic reservoir. Little data regarding the rates of Cryptosporidium infections in ovines in China are available. This study assessed the prevalence of Cryptosporidium spp. in pre-weaned ovines from Aba Tibetan and Qiang Autonomous Prefecture in the Sichuan province of China.
METHODSA total of 213 fecal samples were collected from pre-weaned ovines and were examined microscopically (following modified acid fast staining). In addition, 18S rRNA genetic sequences were amplified from fecal samples by nested PCR and phylogenetically analyzed.
RESULTSThe prevalence of Cryptosporidium in the collected samples was at 14.6% (31/213) and four isolates identified by PCR belonged to the Cryptosporidium cervine genotype (Cryptosporidium ubiquitum) demonstrating that this species was the primary sheep species found in sheep in China.
CONCLUSIONThe present study suggested that the high incidence of Cryptosporidium in sheep poses a significant public health threat and that surveillance practices must be established to prevent zoonotic disease of humans.
Animals ; China ; Cryptosporidium ; genetics ; isolation & purification ; Feces ; parasitology ; Oocysts ; microbiology ; Polymerase Chain Reaction ; Sheep ; Weaning
7.Hemolytic disease of fetus and newborn caused by anti-Di a: a case report
Yuanjun WU ; Yong YANG ; Yanli JI ; Chunyan MO ; Zhongying XIE ; Qianquan YUAN ; Jiajun ZENG ; Miaozhen WEN ; Shujie WU
Chinese Journal of Perinatal Medicine 2020;23(11):778-781
We report a case of a newborn baby who suffered from hemolytic disease of fetus and newborn (HDFN) caused by anti-Di a. The baby presented with worsening jaundice started at three hours after birth and was transferred to Dongguan Maternal and Child Health Care Hospital. The newborn's hemoglobin (Hb) was 82 and 76 g/L at five and nine hours after birth, and the total bilirubin (TBIL) was 243.2 and 309.8 μmol/L, respectively. Blood samples of the newborn and the parents were collected for HDFN immunohematology test twelve hours after birth. They showed that the newborn and the father's blood type was A and RhDCCee, while the mother was A and RhDCcee. Direct antiglobulin test (DAT) indicateda strong positive for the newborn and negative for the parents. The reaction of the reagent to red blood cells for antibody screening with the patient's plasma, red cells eluate, and the mother's plasma were all negative, but were positive with the father's red blood cells. The newborn was recovered after treating with phototherapy, intravenous immunoglobulins and urgent blood exchange (the exchanged blood was the same ABO and RhD blood type and cross-matched). The newborn's plasma and red cells eluate were collected before blood exchange, and the mother's plasma were used to assess the red blood cells reaction, and IgG anti-Di a was identified in each sample. Di a blood typing was positive for the newborn and the father, and negative for the mother. Therefore, the newborn was diagnosed as HDFN caused by anti-Di a.
8.Cyclospora cayetanensis infections among diarrheal outpatients in Shanghai: a retrospective case study.
Yanyan JIANG ; Zhongying YUAN ; Guoqing ZANG ; Dan LI ; Ying WANG ; Yi ZHANG ; Hua LIU ; Jianping CAO ; Yujuan SHEN
Frontiers of Medicine 2018;12(1):98-103
Cyclospora cayetanensis is a foodborne and waterborne pathogen that causes endemic and epidemic human diarrhea worldwide. A few epidemiological studies regarding C. cayetanensis infections in China have been conducted. During 2013, a total of 291 stool specimens were collected from patients with diarrhea at a hospital in urban Shanghai. C. cayetanensis was not detected in any of the stool specimens by traditional microscopy, whereas five stool specimens (1.72%, 5/291) were positive by PCR. These positive cases confirmed by molecular technology were all in the adult group (mean age 27.8 years; 2.94%, 5/170) with watery diarrhea. Marked infection occurred in the rainy season of May and July. Sequence and phylogenetic analyses of the partial 18S rRNA genes of C. cayetanensis isolated showed intra-species diversity of this parasite. This study showed, for the first time, that C. cayetanensis is a pathogen in outpatients with diarrhea in Shanghai, albeit at a low level. However, the transmission dynamics of this parasite in these patients remain uncertain.
Adolescent
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Adult
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China
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epidemiology
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Cyclospora
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genetics
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isolation & purification
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Cyclosporiasis
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epidemiology
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Diarrhea
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etiology
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parasitology
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Feces
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parasitology
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Female
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Humans
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Male
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Middle Aged
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Outpatients
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Phylogeny
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Polymerase Chain Reaction
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RNA, Ribosomal, 18S
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analysis
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Retrospective Studies
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Young Adult