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Qualitative and quantitative analysis of 2-(2-phenylethyl) chromones in sodium chloride(NaCl)-treated suspension cells of Aquilaria sinensis was conducted by UPLC-Q-Exactive-MS and UPLC-QQQ-MS/MS. Both analyses were performed on a Waters T3 column(2.1 mm×50 mm, 1.8 μm) with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as mobile phases at gradient elution. MS data were collected by electrospray ionization in positive ion mode. Forty-seven phenylethylchromones was identified from NaCl-treated suspension cell samples of A. sinensis using UPLC-Q-Exactive-MS, including 22 flindersia-type 2-(2-phenylethyl) chromones and their glycosides, 10 5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones and 15 mono-epoxy or diepoxy-5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones. Additionally, 25 phenylethylchromones were quantitated by UPLC-QQQ-MS/MS. Overall, the rapid and efficient qualitative and quantitative analysis of phenylethylchromones in NaCl-treated suspension cells of A. sinensis by two LC-MS techniques, provides an important reference for the yield of phenylethylchromones in Aquilariae Lignum Resinatum using in vitro culture and other biotechnologies.
Subject(s)
Chromones , Sodium Chloride , Chromatography, Liquid , Flavonoids , Tandem Mass Spectrometry , ThymelaeaceaeABSTRACT
Cytochrome P450 reductase (CPR) is essential for the electron transport chain of cytochrome P450s, playing an indispensable role in electron transfer in vivo. In this study, one cDNA encoding cytochrome P450 reductase (Ascpr1) was identified from the callus of Aquilaria sinensis. Ascpr1 contains an open reading frame of 2 124 bp. The deduced protein is composed of 707 amino acids, with a predicted molecular weight of 78.82 kD. Phylogenetic analysis revealed that AsCPR1 is a type Ⅱ CPR protein closely related to the CPR from Theobroma cacao. Transmembrane prediction using TMHMM 2.0 indicated that the amino acids 52-71 of AsCPR1 comprise a transmembrane region. After truncating of 67 amino acid residues from N-terminal, the truncated AsCPR1 was successfully expressed in E. coli Transetta (DE3). Further purification of the recombinant AsCPR1 by affinity chromatography and determination of the enzymatic activity allowed the reducing ability of AsCPR1 to cytochrome C in vitro. The results pave the way for further study on the synthesis of defensive chemicals involved in P450s and the functions of CPR in self-defense of A. sinensis.
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In the central nervous system, hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are essential to maintain normal neuronal function. Recent studies have shown that HCN channels may be involved in the pathological process of ischemic brain injury, but the mechanisms remain unclear. Autophagy is activated in cerebral ischemia, but its role in cell death/survival remains controversial. In this study, our results showed that the HCN channel blocker ZD7288 remarkably decreased the percentage of apoptotic neurons and corrected the excessive autophagy induced by oxygen-glucose deprivation followed by reperfusion (OGD/R) in hippocampal HT22 neurons. Furthermore, in the OGD/R group, p-mTOR, p-ULK1 (Ser), and p62 were significantly decreased, while p-ULK1 (Ser), atg5, and beclin1 were remarkably increased. ZD7288 did not change the expression of p-ULK1 (Ser), ULK1 (Ser), p62, Beclin1, and atg5, which are involved in regulating autophagosome formation. Besides, we found that OGD/R induced a significant increase in Cathepsin D expression, but not LAMP-1. Treatment with ZD7288 at 10 μmol/L in the OGD/R group did not change the expression of cathepsin D and LAMP-1. However, chloroquine (CQ), which decreases autophagosome-lysosome fusion, eliminated the correction of excessive autophagy and neuroprotection by ZD7288. Besides, shRNA knockdown of HCN2 channels significantly reduced the accumulation of LC3-II and increased neuron survival in the OGD/R and transient global cerebral ischemia (TGCI) models, and CQ also eliminated the effects of HCN2-shRNA. Furthermore, we found that the percentage of LC3-positive puncta that co-localized with LAMP-1-positive lysosomes decreased in Con-shRNA-transfected HT22 neurons exposed to OGD/R or CQ. In HCN2-shRNA-transfected HT22 neurons, the percentage of LC3-positive puncta that co-localized with LAMP-1-positive lysosomes increased under OGD/R; however, the percentage was significantly decreased by the addition of CQ to HCN2-shRNA-transfected HT22 neurons. The present results demonstrated that blockade of HCN2 channels provides neuroprotection against OGD/R and TGCI by accelerating autophagic degradation attributable to the promotion of autophagosome and lysosome fusion.
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Objective To understand the knowledge, attitude and practice (KAP) of foodborne parasitic diseases among medical college students, so as to provide the evidence for the development of strategies on health education. Methods A total of 929 premed students from a medical college were selected in Hebei Province using the stratified sampling method from October to December, 2017, and investigated by questionnaires to understand their KAP on foodborne parasitic diseases. Results The understanding and concern degree on foodborne parasitic diseases among the college students from different majors were statistically different (all P < 0.05), the awareness rate and the degree of concern of the non-medical students were lower than those of the medical students, and the awareness rate and the degree of concern of the students who had received medical parasitology education were higher than those who had not received. For the formation rate of not prefer seeking novelty, there was a statistically significant difference among the college students in different majors (P < 0.05), and the rate of health behavior formation in non-medical students was relatively low. There were statistically significant differences among the students from different majors in the attitude on "would suggest others not to eat the foods that might be infected with the foodborne parasites", "would not eat the foods that might be infected with the foodborne parasites", and "would give up special preferences or bad habits" (all P < 0.05), and the formation rate of correct attitude of medical students was relatively high. Conclusion The awareness rate and the formation rates of correct attitude and behavior of the college students who have received medical parasitology education are relatively higher, so it is necessary to set up a course on foodborne parasitic diseases in colleges and universities.
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Objective To understand the knowledge, attitude and practice (KAP) of foodborne parasitic diseases among medical college students, so as to provide the evidence for the development of strategies on health education. Methods A total of 929 premed students from a medical college were selected in Hebei Province using the stratified sampling method from October to December, 2017, and investigated by questionnaires to understand their KAP on foodborne parasitic diseases. Results The understanding and concern degree on foodborne parasitic diseases among the college students from different majors were statistically different (all P < 0.05), the awareness rate and the degree of concern of the non-medical students were lower than those of the medical students, and the awareness rate and the degree of concern of the students who had received medical parasitology education were higher than those who had not received. For the formation rate of not prefer seeking novelty, there was a statistically significant difference among the college students in different majors (P < 0.05), and the rate of health behavior formation in non-medical students was relatively low. There were statistically significant differences among the students from different majors in the attitude on "would suggest others not to eat the foods that might be infected with the foodborne parasites", "would not eat the foods that might be infected with the foodborne parasites", and "would give up special preferences or bad habits" (all P < 0.05), and the formation rate of correct attitude of medical students was relatively high. Conclusion The awareness rate and the formation rates of correct attitude and behavior of the college students who have received medical parasitology education are relatively higher, so it is necessary to set up a course on foodborne parasitic diseases in colleges and universities.
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@#【Objective】To explore the clinical characteristics of aortic dissection involved in coronary artery in order to avoid misdiagnosis，mistreatment，and take correct treatment methods in time.【Methods】Twelve cases of aortic dissection（AD）manifesting as acute myocardial infarction（AMI）were analyzed retrospectively and followed up.【Results】 A total of 12 cases were enrolled from 288 cases of AD patients，of which 2 cases（16.7%）were inferior wall myocardial infarction，1 cases（8.3%）were anterior wall myocardial infarction，9 cases（75%）were non ST segment elevation myocardial infarction，male patients was significantly predominant over female，leading to the male/female ratio of 3∶1，the age was 43~76（56.9±12.2）years. Up to 66.7% of patients had a history of hypertension. 11 patients（91.7%）were admitted to hospital for different degrees of chest pain，with back pain in 9 cases（75%），syncope in 4 cases（33.3%），hypotension in 6 cases（50%），and irritability in 7 cases（58.3%）. All the myocardial enzymes were positive in the laboratory and significantly elevated D-dimer；3 cases were performed coronary arteriography but not found the opening of coronary artery. AD was confirmed by transthoracic echocardiography and chest CTA. 3 cases of surgery were performed，1 cases failed and died within 24 hours，and 2 cases were successful. The average follow-up 5 hours to 12 months，2 cases survived and the remaining 10 cases died.【Conclusions】Myocardial infarction caused by aortic dissection is rare in clinic ，but it has critical and high mortality rate，and it is easily misdiagnosed，with poor prognosis. Early diagnosis and timely operation can improve the prognosis. In the case of AMI with back pain，irritability，and abnormal rise of D two polymer，the disease should be highly suspected.
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OBJECTIVE@#To analyze the risk factors of preoperative upper respiratory infections in children with cleft lips and palate (CLP) and investigate preventive measures to reduce infections and improve the quality of treatments.@*METHODS@#A total of 510 children with CLP of ages 3 years old or younger were selected from hospital cases from June to December 2017. The test group comprised 50 children with upper respiratory infections, whereas the control group comprised 460 children without upper respiratory infections. A t-test and a multivariate logistic analysis were utilized to analyze the risk factors and to investigate the preventive measures.@*RESULTS@#Feeding patterns, the presence of infected companions during hospitalization, and ventilation at night were statistically significant. The feeding patterns and the presence of infected companions during hospitalization were independent risk factors for upper respiratory infections in children with CLP.@*CONCLUSIONS@#Bottle feeding, infected companions during hospitalization, and the absence of window ventilation at night are risk factors for preoperative upper respiratory infections in children 3 years old or younger with CLP. Among the risk factors identified, feeding patterns and the presence of infected companion during hospitalization were the most influential. Medical staff members should streng-then corresponding health education and nursing measures to control the risk factors.
Subject(s)
Child , Child, Preschool , Humans , Cleft Lip , General Surgery , Cleft Palate , General Surgery , Cross Infection , Risk FactorsABSTRACT
Objective: To assess fluid-attenuated inversion recovery (FLAIR) vascular hyper-intensity (FVH) and explore its relationship with CT perfusion (CTP) penumbral/infarct core mismatch ratio and diffusion weighted imaging (DWI) final infarct volume in acute ischemic stroke (AIS) patients with middle cerebral artery occlusion (MCAO). Methods: The CTP and MRI images of 38 AIS patients with MCAO were reviewed. The FVH score (longitudinal direction) [FVH score (L)] and FVH score (transverse direction) [FVH score (T)] were quantified on the FLAIR images. The FVH score (L) (range, 0-16) was based on a rostrocaudal extension of FVH and the FVH score (T) (range, 0-3) was based on FVH supply of the occluded MCA territory. The mismatch ratio was calculated from the ratio of the [mean transit time - cerebral blood volume (CBV)] lesion/CBV lesion on the CTP images. The DWI infarct volume was measured on the DWI images. Results: The mismatch ratio was larger for the group of FVH score (L)=7~8 than those of FVH score (L)=5~6 and FVH score (L)=3~4 (p=0.03), whereas the DWI infarct volume was smaller (p=0.04). Similarly, the mismatch ratio of FVH score (T)=2~3 group was larger than FVH score (T)=1 group (p=0.01), whereas the DWI infarct volume was smaller (p=0.02). Both FVH score (L) and FVH score (T) correlated positively with mismatch ratio (P=0.02, P=0.001, respectively), but negatively with DWI infarct volume (P=0.03, P=0.004, respectively). Conclusions: Higher FVH score is associated with larger mismatch ratio and smaller DWI infarct volume in AIS patients with MCAO. FLAIR vascular hyperintensity may represent collateral arterial circulation, and may play a role in protecting the ischemic penumbra.
Subject(s)
Infarction, Middle Cerebral ArteryABSTRACT
Objective: To study the mechanism of Wnt/β-catenin signaling pathway in the enhanced malignant phenotype of A549 cells of human non-small cell lung cancer induced by the anti-angiogenesis therapy. Methods: The siRNA technique was employed to inhibit the expression of vascular endothelial growth factor (VEGF) in A549 cells and simulate the clinical course of anti-angiogenesis therapy. Real-time PCR and western blot were used to study the change in the expression of Wnt/β-catenin signaling molecules at the mRNA and protein level respectively, as well as the effect on the epithelial mesenchymal transition in A549 cells. The proliferation and invasion abilities of tumor cells were detected to discuss the mechanism of Wnt/β-catenin signaling pathway in the enhanced malignant phenotype of non-small cell lung cancer induced by the anti-angiogenesis therapy. Results: The specific siRNA could significantly inhibit the expression of VEGF in cells to simulate the anti-angiogenesis therapy. Under the action of 50 nM VEGF siRNA, the proliferation ability of A549 significantly increased (P < 0.05). After being treated with VEGF siRNA, the invasion ability of cells increased. Twenty-four hours after the transcription of 50 nM siRNA into cells, the number of cells that come through the membrane was 278.3 ± 12.9. Compared with the Ctrl siRNA group, when VEGF was inhibited, the expression of β-catenin and Cyclin D1 increased by 86% and 55% respectively. Meanwhile, the expression of E-cadherin decreased, while the one of vimentin increased. Conclusions: siRNA can significantly inhibit the expression of VEGF. For the anti-angiogenesis therapy, the inhibited expression of VEGF can activate the Wnt/β-catenin signaling pathway to cause the epithelial mesenchymal transition and then the enhanced malignant phenotype of non-small cell lung cancer.
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OBJECTIVE: To determine the regulating role of phosphatidylinositol 3-kinase( PI3K) / protein kinase B( Akt)signaling pathway in the autophagy activity of rat NR8383 cells exposed to silicon dioxide( SiO_2). LY294002 was used to block PI3 K pathway. METHODS: i) The normal NR8383 cells were used and divided into blank group and silica exposure group( final concentrations of SiO_2 suspension were 0 and 50 mg / L respectively). They were cultured for 3,6,12,20 and24 hours. The enzyme linked immunosorbent assay( ELISA) was used to assess the amount of tumor necrosis factor-α( TNF-α) and transforming growth factor-β1( TGF-β1) in supernatants of cultured cells,and then the optimal time of cells exposed to dust was determined. ii) NR8383 cells were divided into control group( treated with a same volume of F-12 K medium without serum),silica group( treated with SiO_2 suspension,final concentration 50 mg / L) and intervention group( treated with SiO_2 suspension and PI3 K inhibitor LY294002,final concentration 50 mg / L and 20 μmol / L,respectively).Cells were harvested following incubation. ELISA was used to detect the levels of TNF-α and TGF-β1 at the time point of20 hours after incubation. To reveal the autophagy status of cells,Western blotting was used to detect Akt and microtubuleassociated proteins 1 light chain 3( LC3) protein at time point of 20 hours; laser scanning confocal microscope( LSCM)was used to observe the immunofluorescence expression of autophagy at time points of 3,6,12 and 20 hours. The cells were also treated with the lysosomal inhibitor chloroquine diphosphate( CDP) at the same time of SiO_2 treatment. RESULTS: i) The time point of 20 hours was confirmed to be the best dust exposure time for in vitro cell model of NR8383 cells.ii) The levels of TNF-α and TGF-β1 of supernatant in the silica group were higher than those of the control group( P <0. 05). The levels of TNF-α and TGF-β1 of supernatant in the intervention group were higher than those of the control group and silica group( P < 0. 05). The Akt protein expression of the intervention group was lower than those in the control group and the silica group,respectively. The LC3 Ⅱ / Ⅰ protein level of the silica group was higher than those of the control group and intervention group( P < 0. 05),but no statistical significance was found between the control group and intervention group( P > 0. 05). LSCM results indicated that autophagy expression at time points of 3 and 6 hours were stronger than those of 12 and 20 hours in control group; autophagy expression at time point of 12 hours was stronger than those of 3 and 6 hours in the silica group,while the autophagy expression at time point of 20 hours was slightly weaker than that of 12 hours,but still stronger than those of 3 and 6 hours. Compared with the same time point in control group,autophagy expression at 3 and 6 hours were weaker in the silica group,while the expressions increased obviously at time points of 12 and 20 hours. Autophagy expression at all time points decreased in the intervention group compared with silica group,especially at the time point of 20 hours. The autophagy expression in each group increased in varying degrees after added with CDP blocking. CONCLUSION: Silica dust exposure can induce autophagy in rat NR8383 cells. PI3 K inhibitor LY294002 can reduce the autophagy expression indicating that the PI3 K / Akt signaling pathway might participate in the autophagy process of silica dust inducing autophagy in alveolar macrophages.
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Salvia miltiorrhiza is one of the most common traditional Chinese medicines. It has rich resources in China. According to modern studies, phenolic acids are the main effective components in S. miltiorrhiza. These components have cardiovascular and cerebrovascular protective effect, and anti-tumor, antioxidant, anti-inflammatory, and antifibrotic activities, etc. It has been widely used for the treatment of cardiovascular and cerebrovascular diseases and others. In this paper, the chemicals and pharmacological effects of phenolic acids from S. miltiorrhiza were summarized in the last decade. Its researches and development prospects were also analyzed for further studying and comprehensive utilization of these phenolic acids.
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OBJECTIVE@#To study the mechanism of Wnt/β-catenin signaling pathway in the enhanced malignant phenotype of A549 cells of human non-small cell lung cancer induced by the anti-angiogenesis therapy.@*METHODS@#The siRNA technique was employed to inhibit the expression of vascular endothelial growth factor (VEGF) in A549 cells and simulate the clinical course of anti-angiogenesis therapy. Real-time PCR and western blot were used to study the change in the expression of Wnt/β-catenin signaling molecules at the mRNA and protein level respectively, as well as the effect on the epithelial mesenchymal transition in A549 cells. The proliferation and invasion abilities of tumor cells were detected to discuss the mechanism of Wnt/β-catenin signaling pathway in the enhanced malignant phenotype of non-small cell lung cancer induced by the anti-angiogenesis therapy.@*RESULTS@#The specific siRNA could significantly inhibit the expression of VEGF in cells to simulate the anti-angiogenesis therapy. Under the action of 50 nM VEGF siRNA, the proliferation ability of A549 significantly increased (P < 0.05). After being treated with VEGF siRNA, the invasion ability of cells increased. Twenty-four hours after the transcription of 50 nM siRNA into cells, the number of cells that come through the membrane was 278.3 ± 12.9. Compared with the Ctrl siRNA group, when VEGF was inhibited, the expression of β-catenin and Cyclin D1 increased by 86% and 55% respectively. Meanwhile, the expression of E-cadherin decreased, while the one of vimentin increased.@*CONCLUSIONS@#siRNA can significantly inhibit the expression of VEGF. For the anti-angiogenesis therapy, the inhibited expression of VEGF can activate the Wnt/β-catenin signaling pathway to cause the epithelial mesenchymal transition and then the enhanced malignant phenotype of non-small cell lung cancer.
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Objective To investigate effects of orthoses and relative techniques applied in burn patients.Methods 20 customized orthoses used for 17 burn cases since the year 2008 were studied, and the effects and main technical points of the treatment were summarized based on the principle of such orthoses. The burn areas in these patients were at the neck, hand, ankle, foot and body trunk. The intervention time of using orthoses was at early stage of burn, recovery stage and late stage of rehabilitation, respectively. Results The review study showed that fitting the orthoses after burn could prevent the contracture of the joint, restrain scar casting, assist training to improve the range of motion of the joint. Conclusions According to the state of the burn patients, the use of well fitted orthoses could have obvious good clinic effect, which promotes the functionality recovery.
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<p><b>OBJECTIVE</b>To investigate the association of the intercellular adhesion molecule-1 gene (ICAM-1) K469E polymorphism and rheumatoid arthritis (RA).</p><p><b>METHODS</b>Two hundred and seventy-five patients with RA and 254 healthy individuals were collected and enrolled in the study. The K469E polymorphism of ICAM-1 gene was analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>The genotype frequencies of KK, KE and EE of K469E polymorphism were 0.535,0.411 and 0.054 respectively in the RA patients, and 0.512,0.437 and 0.051 respectively in the healthy individuals, and there was no significant difference between the two groups (chi² = 0.371, P = 0.831). The frequencies of the K469 allele were 0.74 and 0.73 in the RA patients and the controls respectively (chi² = 0.127, P = 0.721, OR = 1.051, 95%CI:0.800-1.381). No significant difference was observed in KK + KE genotype frequencies between the two groups (P = 0.863), with an odds ratio of 0.935 (95%CI:0.436-2.005).</p><p><b>CONCLUSION</b>The K469E polymorphism of the ICAM-1 gene was not associated with the susceptibility of rheumatoid arthritis.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Arthritis, Rheumatoid , Genetics , Disease Susceptibility , Genotype , Intercellular Adhesion Molecule-1 , Genetics , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment LengthABSTRACT
<p><b>OBJECTIVE</b>To evaluate the clinical significances of combined detections of the autoantibodies ANA, ANAs and anti-dsDNA in patients with systemic lupus erythematosus (SLE).</p><p><b>METHODS</b>ANA was tested by indirect immunofluorescent method (IIF), ENA was tested by Western blot and anti-dsDNA was measured with radioimmunoassay in the serum samples from patients with SLE (n=90), and from the comparison group (n=74) and healthy control group (n=53).</p><p><b>RESULTS</b>The sensitivity of anti-dsDNA, AnuA,anti-Sm in patients with SLE was 41.1%, 32.2% and 22.2%. The specificity of anti-dsDNA, AnuA, anti-Sm in SLE patients was all 97.3%. The positive likelihood ratios were 15.22, 11.93 and 8.22, respectively. The sensitivity and specificity reached 62.2% and 90.5%, respectively by combined determinations of anti-dsDNA, AnuA and anti-Sm.</p><p><b>CONCLUSION</b>The combined examinations of anti-dsDNA, AnuA, anti-Sms can improve the sensitivity and efficiency of laboratory diagnosis of SLE.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Antibodies, Antinuclear , Blood , Autoantibodies , Blood , Lupus Erythematosus, Systemic , Diagnosis , Allergy and Immunology , Sensitivity and SpecificityABSTRACT
Objective To analysis and determine the possibility of the Citellus undulatus infected with Yersinia pestis surviving the winter in an experimental study, and to provide scientific experimental basis for the study on the mechanism of Yersinia pestis preservation. Method In 2006,09 to 2007,04 and 2007,09 to 2008,04 in Xinjiang Wusu-Gurtu natural foci of plague, under natural conditions, the over the winter process of Citellus undulatus carrying the plague bacteria was simulated, and 178 Citellus undulatus were infected with Yersinia pestis (1×107 Bacteria/mouse) using artificial injection method. One hundred seventy-eight Citellus undulatus infected with Yersinia pestis were kept into a construction of the black (1-5 ℃) basement (2 meters under the ground) in the plague focus. In doing so, these Citellus undulatuses almost simultaneously stepped into hibernation. After waking up from hibernation in following year in April, the survived mice carrying the plague bacteria were observed. Results Sixty-eight mice survived among the 178 infected with Yersinia pestis after 6 months of hibernation (through October to the following year in April), and the remaining 110 were all dead without pulling through the hibernation period. The survival rate was 38.2% (68/178). The organ culture of Yersinia pestis of the 110 dead mice(Citellus undnlatus) were tested, 67 were negative(-), 43 positive(+), with a positive rate of 39.1%(43/110). Among the rats with positive plague bacteria, the congestive pulmonary edema and the pathological changes of the hemorrhagic inflammation of the heart, liver, spleen, kidney and injection site could be seen clearly; the plague-free mice were not found to have any pathological changes. The survived 68 mice over the winter were autopsied and observed after being fed up for 20 days. No any pathological changes were found among these mice, and culturing of Yersinia pestis of the heart, liver, spleen, lungs and the tissue of injection site of these mice were all negative (-). Conclusions Citellus undulatus can carry Yersinia pestis during hibernation, but some fail to carry the bacteria through the entire process of hibernation persistently. Yersinia pestis was negative in the survived mice at the end of hibernation. The results showed that Citellus undulatus can not carry Yersinia pestis over the winter.