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Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.
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Objective:To investigate the effects of different chimerism strategies and different immune ways on the two antigen-dominant regions of Xinjiang hemorrhagic fever virus (XHFV) glycoprotein.Methods:The 5' end was added or not added with interleukin-2 (IL-2) signal peptide and the general-purpose auxiliary T cell epitopes as different design strategies. GcⅠ and GcⅡ and the epitopes previously identified on GcⅠ (Gc 233-248, Gc 241-256 and Gc 281-296) were fused and constructed into the eukaryotic expression vector pVAX1 and the prokaryotic expression vector pET-28a. The recombinant prokaryotic plasmid transformed into E.coli BL21 was induced and purified, and the recombinant eukaryotes were extracted by indirect immunofluorescent assay. BALB/c mice were immunized by protein immunity, gene immunity, and DNA prime-protein boost immunity. The IgG antibody level was measured by ELISA. The immune effect was evaluated by the proliferation of T-lymphocytes and the content of cytokines in the spleen. Results:The results of double enzyme digestion and sequencing showed that eight recombinant plasmids were successfully constructed, and the recombinant eukaryotes were successfully expressed in vitro by fluorescence microscopy. After three times of immunization, the IgG level and the proliferation of T-lymphocytes in the spleen of mice in the experimental group were significantly higher than those in the control group ( P<0.01). The mass concentration test results of Th2 cytokines IL-4 and Th1 cytokines interferon-gamma (IFN-γ) revealed that the response of the DNA prime-protein boost immunity was biased to Th1. Conclusions:The multi-epitope chimeric vaccine of XHFV glycoprotein was successfully constructed, and the target antigen could be expressed effectively in vivo. The immune groups stimulated stronger humoral and cellular immune responses compared with the control group. Among them, the immune effect of pVAX1-ST(GcⅠe+GcⅡ) combined with recombinant protein r(GcⅠe+GcⅡ) was the best, and it is expected to be a new candidate vaccine for XHFV.
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Objective:To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli ( E. coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. Methods:Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. Results:Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass ( Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×10 3, 37.1×10 3, 31.9×10 3, 30.8×10 3, 40×10 3 and 54.4×10 3, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. Conclusions:The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines.
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Objective To express and purify two domains GcⅠand GcⅡof Xinjiang hemorrhagic fever virus (XHFV) glycoprotein, and to study its immunogenicity and the effects on immune response in mice. Methods The prokaryotic expression plasmids of pET28a-GcⅠand pET32a-GcⅡwere constructed and transformed into E. coli BL21, respectively. The expression and purification conditions of rGcⅠand rGcⅡproteins were optimized. The antigenicity of the fusion protein was detected by Western Blot and enzyme-linked immunosorbent assay (ELISA). BALB/c mice were immunized by protein immunization and DNA priming-protein boosting. The mice were randomly divided into 5 groups, including pVAX1-GcⅠ+rGcⅠgroup, pVAX1-GcⅡ+rGcⅡgroup, rGcⅠgroup, rGcⅡgroup and saline group (control group) with 7 mice in each group. The serum antibody titer of mice was detected by indirect ELISA, and the immune effect was evaluated by spleen T lymphocyte proliferation assay and cytokine content determination. Results The fusion proteins rGcⅠand rGcⅡwere purified and obtained, which could react with positive serum of sheep and had good antigenicity. After three immunizations, the IgG levels in the serum of each experimental group were significantly higher than those in the control group (all P<0.001). The serum antibody titers of the experimental groups were reached above 1:12800. Among them, the concentration of Th2 type cytokine interleukin-4 (IL-4) in the spleen cell culture supernatant of rGcⅡ[(79.97±7.47) ng/L] and pVAX1-GcⅡ+rGcⅡgroup [(61.43±9.27) ng/L] was significantly higher than (24.29±3.81) ng/L of the control group, respectively (all P<0.01). The highest mass concentration [(42.46 ±2.60) ng/L] of Th1 type cytokine interferon-γ(IFN-γ) was observed in the pVAX1-GcⅡ+rGcⅡ group, which was significantly higher than (20.33±1.67) ng/L of the control group, and the difference was statistically significant (P<0.001). That showed a significant antigen-specific splenic T lymphocyte proliferation (P<0.001). Conclusions The purified recombinant proteins rGcⅠand rGcⅡhave good immunogenicity, which can make the immune system T lymphocytes tend to Th2 response, and pVAX1-GcⅡ combined with recombinant protein GcⅡ can induce better antigen-specific immune effect. And pVAX1-GcⅡ combined with recombinant protein GcⅡis expected to be used as vaccine candidates for the prevention and control of XHFV.
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The disease caused by Rickettsia is an important zoonotic disease. Due to the complexity of the ecological environment of Xinjiang, the vectors, host and species of Rickettsia are complex and diverse; its natural foci are widely distributed, so they have brought great harm to local human health and livestock production. With global warming, rapid development of social economy and promotion of foreign economic and trade exchanges in Xinjiang, these may become potential factors for the diversity of Rickettsia vectors and hosts. In recent years, some rickettsial disease such as the onset of Q fever, ehrlichiosis and anaplasmosis, are increased at some areas of Xinjiang, thus, the Rickettsia and rickettsial diseases have been widely concerned. At present, a variety of new Rickettsia species have been detected and isolated from several areas of Xinjiang, and their public health significance have also been paid more and more attention. In this paper, we briefly summarized the research status about reported Rickettsia species identification, media host and popular distribution in the Xinjiang region over the past 20 years, to providing scientific basis for a systematic study and diagnosis, prevention and treatment of Rickettsia and rickettsial disease.
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Objective To construct two DNA vaccines based on two glycoprotein antigen segments of Xinjiang hemorrhagic fever virus (XHFV) and to evaluate the immune responses in BALB/c mice following vaccination.Methods Two recombinant expression plasmids pVAX1-GcⅠand pVAX1-Gc Ⅱ were constructed by inserting XHFV YL04057 strain Gc Ⅰ (1 229-1 349 aa) and Gc Ⅱ (1 443-1 566 aa) fragments into the eukaryotic expression vector pVAX1 and then were identify by restriction enzyme digestion and sequencing analysis.The recombinant expression plasmids were transfected into mice by hydrodynamics-based transfection.Immune responses induced in mice were evaluated by testing the proliferation of T cells with MTT,measuring serum antibody level with ELISA and detecting cytokines in the supernatant of spleen cell culture with ELISA kit.Results The recombinant expression plasmids were successfully constructed as indicated by the results of restriction enzyme digestion and sequencing analysis.Expression of Gc Ⅰ and Gc Ⅱ genes in mice liver tissues was detected.Antibody titers in mice immunized with pVAX1-GcⅠor pVAX1-Gc Ⅱ were higher than those in mice immunized with pVAX1.Compared with pVAX1,pVAX1-Gc Ⅱ significantly enhanced the proliferation of splenic T lymphocytes and the expression of IFN-γ (P<0.01).Conclusion The constructed two DNA vaccines for XHFV can induce specific humoral and cellular immune responses in mice.pVAX1-Gc Ⅱ is better than pVAX1-GcⅠin immunogenicity and protective efficacy,suggesting that it can be used as a promising candidate for the development of DNA vaccine for XHFV.
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It is an important topic that the plague vector efficiency has been studied in plague epidemiology from the plague found out,and it is one of the key issues that we recognized the plague preservation and prevalence in nature ecological system too.Current related biological basis,technical theory and new technology research and application,and other fields have made important achievements and progress in the study of plague vector efficiency,but a systematic and standardized evaluation of vector competency and efficiency is needed in modem vector transmission plague research.It is,therefore,necessary to establish specific and valid research indicators based on the plague natural ecological system and the plague nature quality and specific scientific research purpose,and these indicators will also be able to serve the research goal,improve the credibility and comparability of vector efficiency of research results.
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Objective@#To understand the histopathological and ultrastructural pathology changes of great gerbils in the Junggar Basin to Yersinia pestis infection.@*Methods@#Forty captured great gerbils from the Junggar Basin that tested negative for anti-F1 antibodies were infected. The Y. pestis strain 2504, isolated from a live great gerbil in the natural plague foci of the Junggar Basin in 2005 with a median lethal dose (LD50) of <10 CFU/ml, was used in this study. Forty great gerbils were divided into seven infection groups and were subcutaneously infected with 7.4×105, 7.4×106, 7.4×107, 7.4×108, 7.4×109, 7.4×1010, or 3.0×1011 CFU/ml of 2504. One milliliter of physiological saline was injected in the noninfected group as a control. We collected the liver, spleen, heart, and lung from all animals for histopathologic and ultrastructural pathology examination.@*Results@#Great gerbils in the 7.4×108-3.0×1011 CFU/ml groups did not survive and exhibited pathological changes and altered ultrastructural pathology. The liver tissue of infected great gerbils showed spotty necrosis and fatty degeneration, intranuclear canaliculi with increased hepatocytes, and uneven distribution of organelles. Additionally, reactive proliferation of lymphoid tissue in the spleen, blood sinusoid lacunae with neutrophil infiltration, and phagocytosed bacteria in phagocyte cells were observed. Myocardial fiber hypertrophy and interstitial indistinction, nuclear matrices decreased in cardiac myocytes, and loose arrangement of myogenic fibers in myocardial cells were also observed. Angiectasia, capillary congestion, and tissue necrosis were found in the lung. No significant difference in histopathological and ultrastructural pathology in the parenchymal organ was observed between the 7.4×105-7.4×107 CFU/ml groups and the 7.4×108-3.0×1011 CFU/ml groups, and no specific death caused by Y. pestis infection was apparent in the 7.4×105-7.4×107 CFU/ml groups.@*Conclusion@#Y. pestis infection altered tissue and ultrastructural pathology in the parenchyma apparatus of great gerbils. In particular, the liver and spleen appeared to be the primary site of Y. pestis infection in great gerbils.
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Objective@#To observe the dynamics of antibody response in great gerbils infected with Yersinia pestis in experiment.@*Method@#A total of 211 great gerbils were captured in the southern margin of plague natural focus of Junggar Basin of the Xinjiang Uygur Autonomous Region in 2011. Among them, there were 167 great gerbils without infection of Y. pestis and 44 great gerbils infected by Y.pestis. Y.pestis No. 2504 was employed for this experimental strain, which was strong toxic strain with negativity in the reduction experiment of nitrate. 35 great gerbils without the infection of Y. pestis were divided randomly and averagely into 7 groups including 6 experimental groups and 1 control group. Great gerbils in the 1st to 6th experimental groups were exposed first with 1 × 106-1 × 1011 CFU/ml of bacterial fluid with 10 times of gradient dilution; groin areas of great gerbils in the control group were injected subcutaneously with physiological saline; and the amount of infection was all 1 ml. 17 great gerbils infected with Y. pestis and the first detection of F1-antibody titer in 1∶256-1∶4 096 were grouped according to F1-antibody titer: group 1∶4 096 (n=4), group 1∶2 048 (n=4), group 1∶1 024 (n=3), group 1∶512 (n=3) and group 1∶256 (n=3); and blood in caudal regions was collected in asepsis for the detection of F1-antibody, with a total of 5 times. 9 great gerbils which were selected from the remaining great gerbils infected with Y. pestis and detected F1-antibody negative 2 times were exposed 1×106 CFU/ml of bacterial fluid for the second infection, with the amount of infection being 1 ml. Blood in caudal regions of great gerbils after the first and second infection were collected for the detection of plague F1-antibody on the 3rd, 5th, 7th, 15th, 30th, 60th, 90th and 120th day after infection. Declined regression models for great gerbils' antibodies were established with unary linear regression equation; declined change diagrams for the antibodies were drawn to observe the declined F1-antibody after great gerbils were exposed to Y. pestis.@*Results@#In great gerbils with the first infection of Y. pestis, antibodies were detected in the 1 × 106-1 × 108 CFU/ml of group on the 30th, 15th and 15th day, respectively; the positive rates of antibody were 1/4, 3/4 and 4/5, respectively; the group 1×107 and 1× 108 CFU/ml reached to the highest antibody titer with 1∶256 on the 120th day; antibodies were revealed in the group 1×109, 1×1010 and 1×1011 CFU/ml from the 5th to 7th day when the seroconversion of all antibodies was observed; group 1×1011 CFU/ml reached to the highest antibody titer on the 120th day with 1∶4 096. In the great gerbils with the second exposure to Y.pestis, positive antibodies were detected on the 3rd day with the positive rate being 2/9; and the highest antibody titer with 1∶2 048 was noted on the 90th day. Unary linear regression equation of declined F1 antibody of great gerbils was y=0.045x- 0.321 (F=115.40, P< 0.001), and the shortest duration for F1-antibody titer declining from 1∶4 096 to 0 was 140 d and the longest duration 200 d.@*Conclusion@#Great gerbils infected with the high concentration of Y. pestis fluid show shorter duration in producing F1-antibody, the antibody positive rate is also higher, and the highest antibody titer can reach 1∶4 096. The great gerbils could hold the plague F1 antibodies for a long time which was about 140 to 200 days from the highest titer.
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Objective To explore the epidemic situation of animal plague in Junggar Basin natural plague foci.Methods Data on epidemics of plague and on population involved,as well as results on antibodies and pathogens,were analyzed.Samples on animals and vectors were collected from 18 counties in Junggar Basin plague natural foci between 2007 and 2016.Results The density of Rhombomys (R.) opimus was temporally fluctuant,from 2.1/hm2 to 22.6/hm2 respectively.However,the spatial distribution appeared asymmetrical,with the highest seen in Kelamayi and Wumuqimidong counties,as 14.2/hm2 and 13.0/hrn2 respectively.Rates of capture on nocturnal rodents were from 4.2％ to 10.1％,with the highest rate as 10.1％ in 2014.Meriones meridianus appeared the dominant species in the nocturnal community of rodents,which accounted for 81.9％.Regarding the spatial and temporal distributions,rates ofR.opimus with fleas appeared fluctuant,with an average rate as 90.7％ and the average total flea index was 10.44.In flea community of R.opimus,Xenopsylla (X.)skrjabini was found the dominant species,popular in distribution and accounted for 47.8％.The average rate of nocturnal rodents with flea was 20.2％,with total flea index as 1.20 and the dominant fleas were X.conformis conformis and Nosopsyllus laeviceps.A total of 13 species with 9 087 serum samples from rodents were detected as having Y.pestis antibody by IHA,with 617 positive samples.Of them,the positive rate of having R.opimus appeared the highest (9.4％),followed by D.sagitta (1.1％).Spatially,two clustered areas were found,with one in the eastern Junggar Basin from Changji to Mulei county,with the antibody positive rates ofR.opimus as 14.3％.The other one was in the central area of Junggar Basin,including Kelamayi,Shawan and Wusu counties,with the antibody positive rate as 13.6％.The prevalence of plague on R.opimus was fluctuant,with the lowest seen in 2008,with the average antibody positive rate of R.opimus as 1.0％ and the highest as 19.3％ in 2013.A total of 18 strains were isolated from 2007 to 2016.However,most of the strains were isolated from R.opimus and parasitic fleas,accounted for 8/9,in Kelamayi,Wulumuqi-midong and Jimusaer,respectively.Conclusions Complex ecosystem was seen in the Junggar Basin natural plague foci,with the multiple composition of species and different community structure of hosts and vectors,plus the flustering prevalence.Animal plague was seen in the whole region with succession of the plague nature foci.Passive transmission of plague between human beings and animals through close contacts was seen which was driven by economic benefits to some degree.
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Objective To explore the epidemic situation of animal plague in Junggar Basin natural plague foci.Methods Data on epidemics of plague and on population involved,as well as results on antibodies and pathogens,were analyzed.Samples on animals and vectors were collected from 18 counties in Junggar Basin plague natural foci between 2007 and 2016.Results The density of Rhombomys (R.) opimus was temporally fluctuant,from 2.1/hm2 to 22.6/hm2 respectively.However,the spatial distribution appeared asymmetrical,with the highest seen in Kelamayi and Wumuqimidong counties,as 14.2/hm2 and 13.0/hrn2 respectively.Rates of capture on nocturnal rodents were from 4.2％ to 10.1％,with the highest rate as 10.1％ in 2014.Meriones meridianus appeared the dominant species in the nocturnal community of rodents,which accounted for 81.9％.Regarding the spatial and temporal distributions,rates ofR.opimus with fleas appeared fluctuant,with an average rate as 90.7％ and the average total flea index was 10.44.In flea community of R.opimus,Xenopsylla (X.)skrjabini was found the dominant species,popular in distribution and accounted for 47.8％.The average rate of nocturnal rodents with flea was 20.2％,with total flea index as 1.20 and the dominant fleas were X.conformis conformis and Nosopsyllus laeviceps.A total of 13 species with 9 087 serum samples from rodents were detected as having Y.pestis antibody by IHA,with 617 positive samples.Of them,the positive rate of having R.opimus appeared the highest (9.4％),followed by D.sagitta (1.1％).Spatially,two clustered areas were found,with one in the eastern Junggar Basin from Changji to Mulei county,with the antibody positive rates ofR.opimus as 14.3％.The other one was in the central area of Junggar Basin,including Kelamayi,Shawan and Wusu counties,with the antibody positive rate as 13.6％.The prevalence of plague on R.opimus was fluctuant,with the lowest seen in 2008,with the average antibody positive rate of R.opimus as 1.0％ and the highest as 19.3％ in 2013.A total of 18 strains were isolated from 2007 to 2016.However,most of the strains were isolated from R.opimus and parasitic fleas,accounted for 8/9,in Kelamayi,Wulumuqi-midong and Jimusaer,respectively.Conclusions Complex ecosystem was seen in the Junggar Basin natural plague foci,with the multiple composition of species and different community structure of hosts and vectors,plus the flustering prevalence.Animal plague was seen in the whole region with succession of the plague nature foci.Passive transmission of plague between human beings and animals through close contacts was seen which was driven by economic benefits to some degree.
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Objective To explore the clinical manifestations,diagnosed main points,laparoscopic trestment strategy of ectopic appendicitis.Methods The 19 clinical data of ectopic appendicitis,from April 2011 to April 2015 in our hosipital,were retrospective analysised,and we sum up the clinical manifestation,diagnosis and treatment,laparoscopic exploration indications and the technique of laparoscopic appendectomy.Laparoscopy exploration and laparoscopic appendectomy were made on all patients under general anesthesia by tracheal cannula.Results Of 19 patients,16 cases by ultrasonic examination get correct diagnosis before operation,of which the ultrasound diagnosis rate is 92.63％,3 limitations of acute peritonitis patients were preoperative misdiagno~s,they were the right kidney stones,acute cholecystitis and the right side of the annex inflammation;there were 2 patients converted to open operation,and the transfer rate is 10.53％.The blood loss of the operation is about 5-20 mL.They were ambulant after 9 hours,in full flow diet after 12 hours,and the average hospitalization time is 4 days.One patient,no other long-term complications were found postoperatively follow-up of 12 to 18 months,were poke holes in infection,and the complications of laparoscopic surgery is 5.27％.Conclusions Ectopic acute appendicitis has no specific clinical manifestation,and is easy to misdiagnosis.In the auxiliary examination,ultrasound is the first choice for the diagnosis of ectopic appendictis.Laparoscopy exploration and laparoscopic appendectomy are the ideal treatments of emergency treatment of ectopic appendicitis,and are worth of clinic application.
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<p><b>OBJECTIVE</b>To explore the spatial and temporal distributions of animal plague in Junggar Basin natural plague focus.</p><p><b>METHODS</b>Data regarding plague antibody (F1) in serum of Great Gerbil (Rhombomys opimus, R. opimus) which were collected from 2005 to 2012 in Junggar Basin and analyzed. The changing rates on the positivity of F1 that appeared spatially and temporally were also analyzed.</p><p><b>RESULTS</b>A total of 4 825 R. opimus serum samples were collected in 13 administrative regions in Junggar Basin.</p><p><b>RESULTS</b>showed that plague R. opimus existed in two areas-Gurbantonggut desert in the eastern-center and the clay desert of western Junggar Basin. However, in these two areas, the intensity of animal plague prevalence was different. In the former region where Yesinia pestis positive serum was detected from R. opimus, the detected rate of R. opimus was 8.39%. However, in the latter areas, the average positive rate was 1.56%. The changing trends of R. opimus plague prevalence were also varied annually. In the western Junggar Basin, the trend showed a slowly downward profile. The serum positive rate of R. opimus for Yesinia pestis decreased, from 7.59% in 2005 to 0.61% in 2008, and appeared as a resting state that none of the positive sample could be found since then. However, in the eastern-center Junggar Basin area-also named as Gurbantonggut desert which had been divided into 3 segments(western, central and eastern, according to related geographical characteristics), the changing trends of animal plague seemed quite complex. In the western segment, the animal plague had two epidemic peaks-in 2006 and 2010, with the interval of 4 years, with the higher peak of all the three geographic segments as 45.65% in 2010 and the positive serum of R. opimus for plague could be detected each year from 2006 to 2012. However, there were 3 epidemic peaks in the same period in the central and eastern segments. In the central segment, the peaks appeared in 2006, 2009 and 2011, with the intervals as 2.5 years and the average positive rate 8.92% was seen the lowest in Gurbantonggut desert. In the eastern segment, the first 2 peaks appeared the same season as in the central segment, but the third peak appeared in 2012, with the peak interval as 3 years. The positive rate of R. opimus for plague was also different in seasons, with the positive rate higher in autumn than in spring. These findings showed that the animal plague could be continuously prevalent from spring to autumn in the natural foci of plague in the Junggar Basin.</p><p><b>CONCLUSION</b>Both geographical and temporal fluctuations of animal plague existed in the natural foci of Junggar Basin which was also named as geographical heterogeneity. Consequently, animal plague could be divided into two areas-the clay plains desert in the western and the Gurbantonggut desert in the eastern-center Junggar Basin.</p>
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Animals , Gerbillinae , Plague , Epidemiology , Time , Yersinia pestisABSTRACT
<p><b>OBJECTIVE</b>In order to understand the distribution of the host animals in Junggar Basin, this study intended to map the spatial distribution and identifying the risk of Rhombomys opimus in the framework of ecological niche theory based on the "3S" technology.</p><p><b>METHODS</b>Data on Rhombomys opimus was obtained through a series of field surveys. Environmental variables were achieved through data from Remote Sensing. Maxent modeling was built to map the potential distribution of Rhombomys opimus, with its risks identified.</p><p><b>RESULTS</b>The prediction model showed ideal accuracy, with the AUC value as 0.968. Probability of Maximum Youden Index was defined as the threshold being used. The sensitivity and specificity showed as 91.4% and 63.3%, respectively. The accuracy was 73.8%, and the Kappa coefficient was 0.495. The positive predictive value was 59.7%. The negative predictive value was 92.6%. The predicted high risk area was 37 304 km2, with 6.2% in the whole area, distributed in 18 counties, including Changji Hui Autonomous Prefecture, Urumqi, Karamay and so on. The number of people under high risk would come about 120 000, scattering in the areas of 261 square kilometers.</p><p><b>CONCLUSION</b>It was feasible to predict the potential distribution of Rhombomys opimus based on the ecological niche theory as well as environmental variables derived from data through remote sensing. More specific high-risk areas could be identified under this technique so as to guide the monitoring programs.</p>
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The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen.Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nueleoeapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonai serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses.The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.
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Aim To study the infuences of Haishengsu extracted from Tegillarca granosa on cultured tumor cells of lung cancer, breast cancer and carcinoma of stomach from human in vitro and its preventive antitumor effect on transplanted tumor in mice in vivo. Methods The inhibitory effect of Haishengsu on tumor cells proliferation was assayed by MTT method, and transplant tumor models of sarcoma 180 (S 180), Ehrlich ascites carcinoma (EAC), and hepatoma (Heps) were used. Before and after transplantation, different doses of Haishengsu were given to the mice for 10 days. The inhibition rates and life span of the mice were calculated. Results Haishengsu at concentrations of 0.5%～2.0% inhibited the proliferation of tested tumor cells in vitro,especially on lung cancer cells with a dose dependent manner (r= 0.974,P