In order to analyze the immunogenicity of the recombinant EG95s protein ,the recombinant plasmids of pET-1EG95s ,pET-2EG95s and pET-3EG95s which containing respectively 1 ,2 ,and 3 copies EG95s were induced to express HIS-1EG95s ,HIS-2EG95s and HIS-3EG95s ,and then the proteins were purified and identified by western-blotting .The same im-mune process was used ,and 8 weeks-old BALB/c mice were immunized ,then its immunogenicity was analyzed by detecting an-tibody levels in mice by indirect ELISA method .Results showed that for recombinant EG95s proteins after transformation , HIS-1EG95s ,HIS-2EG95s ,and HIS-3EG95s also retained immunogenicity and could induce specific antibodies in mice .One week's late after the first immunization with HIS-1EG95s ,the antibody level of was significantly higher than HIS-2EG95s and HIS-3EG95s .But began from 2 weeks after immunization ,the antibody level of HIS-3EG95s was always higher than that of HIS-1EG95s group during the period of the immune .Both the final antibody titers after immunization of HIS-1EG95s and HIS-2EG95s groups was 1∶819 200 ,while HIS-3EG95s group was 1∶163 840 0 .HIS-1EG95s ,HIS-2EG95s and HIS-3EG95s all induced IFN-γin immune mice ,but the difference was not significant .The HIS-1EG95s showed lower response to Echinococ-cus granulosus positive serum than HIS-2EG95s and HIS-3EG95s .It’s indicated that the HIS-1EG95s and HIS-3EG95s also had good immunogenicity .HIS-3EG95s make recombinant protein immunic effects more lasting ,and benefit to generate more long-lasting protective immunity .This study provides the scientific basis for the immunization of echinococcosis (hydatidosis) .

ObjectiveTo investigate the effects of oxidized low density lipoprotein (oxLDL) on autophagy and its effect on Akt/mTOR/p70S6K signaling pathway in human umbilical vein endothelial cells (HUVECs).MethodsThe cultured HUVECs were divided into either an oxLDL or a control group, and treated with 100 μg/ml oxLDL and equal volume phosphate buffer solution respectively.The cells were collected after 6 h and 12 h.Transmission electron microscopy was used to observe the autophagosome.Real-time fluorescence quantitative polymerase chain reaction was used to detect expression levels of microtubule associated protein 1 light chain 3 (LC3) and p62 mRNA.Western blot was used to detect the expression levels of LC3, p62, P-Akt/Akt, P-mTOR/mTOR, and p-p70S6K/p70S6K.Results Compared with the control group, the number of intracellular autophagosome increased obviously (P<0.05), LC3 mRNA and protein expression levels increased significantly (all P<0.05), and p62 mRNA and protein expression levels decreased significantly (all P<0.05) in the oxLDL group.In addition, the phosphorylated protein expression levels of Akt, mTOR and p70S6K in the oxLDL group were significantly decreased than those in the control group (all P<0.01).However, total protein levels of Akt, mTOR, and p70S6K were not significantly different between the oxLDL group and the control group.Conclusion oxLDL may induce the autophagy of HUVECs via inhibiting the Akt/mTOR/p70S6K signaling pathway.

Objective To study the effect of hydrogen peroxide on microstructure of mice kidney and discuss the toxic effect on mice kidney.Methods Thirty healthy male mice of Kunming Genus were divided into 3 groups at random:control group and two experimental groups. Running water was fed to control group for 10 days while 0.3,3 g/L hydrogen peroxide running water readily prepared was fed to the experimental groups for 10 days. On the 10th day,the kidneys were taken out,and fixed in the fixation solutions,conventionally produced and stained.Finally,they were studied under the optical microscope.Results Experimental groups:in the kidney tissue cytoplasm of proximal convoluted tubule showed hydropic degeneration and vacuolation which depend on dose of hydrogen peroxide.Conclusion Toxic effect on mice kidney can be caused by hydrogen peroxide.

OBJECTIVETo explore effects of 650 nm laser and moxibustion pretreatment on visceral traction pain (VTP) and its mechanism.

METHODSForty male SD rats were randomly devided into a sham operation group (group A), a VTP group (group B), a 650 nm laser pretreatment group (group C), a moxibustion pretreatment group (group D). Rats in group A and group B were not treated except sham operation or VTP model. In group C and D, the VTP models were produced immediate after 650 nm laser irradiation or moxibustion at "Zusanli" (ST 36), respectively. The changes of pain score and systolic pressure were investigated and the activity of AChE, the content of SP and leu-enkephaline (LEK), and the positive index of c-Fos protein and glial fibrillary acidic protein (GFAP) were detected by biochemistry, radio-immunity method and immunohistochemistry, respectively.

RESULTSCompared with group A, the pain score, systolic pressure, the activity of AChE, the content of SP, and the positive index of c-Fos protein and GFAP of group B increased significantly (all P < 0.05); compared with group B, the pain score, AChE activity, the content of SP and the positive index of c-Fos protein and GFAP of both group C and group D decreased significantly (all P < 0.05); compared with group B, the content of LEK increased and systolic pressure decreased significantly in group C (both P < 0.05).

CONCLUSIONBoth 650 nm laser and moxibustion pretreatment can inhibit VTP and the mechanism may be related to reducing the activity of AChE and the content of SP, and increasing the activity of LEK and decreasing the expression of c-Fos protein and GFAP.

Animals ; Combined Modality Therapy ; Enteric Nervous System ; physiopathology ; Humans ; Intestinal Pseudo-Obstruction ; physiopathology ; therapy ; Laser Therapy ; Male ; Moxibustion ; Pain ; physiopathology ; Pain Management ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the correlation between apparent diffusion coefficient (ADC) and Ki-67, a marker of tumor activity, in the diagnosis of gliomas.

METHODSConventional magnetic resonance imaging (MRI), enhanced scanning, and diffusion-weighted imaging were performed in 76 patients with pathologically confirmed gliomas. The ADC values were measured at tumor parenchyma and the corresponding contralateral normal brain. The relatively ADC (rADC) values of the tumor parenchyma were calculated. The correlation of the ADC values with tumor grades was analyzed. The expression of Ki-67 was detected by immunohistochemical staining. The correlation between ADC value and Ki-67 in the diagnosis of gliomas was analyzed.

RESULTSThe ADC values and rADC values of high-grade gliomas were significantly lower than those of low-grade gliomas. The ADC values of tumor parenchyma were inversely associated with the degree of malignancy of the gliomas (r=-0.898, r=-0.868; P<0.01). The expression of Ki-67 was significantly higher in high-grade gliomas than that in low-grade gliomas. The Ki-67 labeling index in grade 3 and 4 gliomas were (29.48 ± 19.78)% and (31.21 ± 17.50)%, respectively. Both of them were significantly higher than Ki-67 labeling index in low-grade (grade 1 and 2) gliomas [(2.33 ± 2.20)%] (P<0.01). Nevertheless, the Ki-67 labeling index showed no significant difference between grade 3 and 4 gliomas (P>0.05). The expression of Ki-67 was negatively correlated with the ADC values and rADC values in tumor parenchyma (r=-0.627, r=-0.607; P<0.01).

CONCLUSIONThe ADC and rADC values of tumor parenchyma can indirectly reflect the proliferation and malignancy of gliomas and therefore can be useful for the grading of glioma.

Adolescent ; Adult ; Brain Neoplasms ; diagnosis ; metabolism ; pathology ; Child ; Child, Preschool ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Glioma ; diagnosis ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Young Adult

Objective To study the effect of physical activities and dietary intervention on metabolic syndrome (MS) in primary school students.Methods Either one-year physical activities (PA) or dietary intervention was conducted in Grade 1-5 children from eight primary schools in Haidian district,Beijing.A ‘happy 10 minutes' program was held in the PA group,while the dietary group receiving nutrition lectures.Baseline and post-intervention data on height,weight,waist circumference,serum lipids,glucose,and blood pressure were collected.Results The prevalence of MS at baseline was 9.0％.After intervention programs were carried out,improvements on triglyceride,high density lipoprotein,fasting blood glucose,diastolic blood pressure and waist circumference in the dietary group or PA group were seen,with the dietary group showed better effects than the PA group.The prevalence of MS decreased from 10.4％ to 4.6％ in the dietary group with statistically significant difference,while it increased in both the PA group and the control group.Girls showed better effects in the dietary group.Older students showed better effects than the younger students in the PA group.In non-overweight and overweight students of the dietary intervention group,the MS related components were significantly improved.Conclusion The prevalence of metabolic syndrome was relatively high in the primary school students in Beijing.Education on nutrition could reduce the prevalence and improve the related components,which seemed to be more effective than in the PA intervention.Sex,age and nutritional status were the confounding factors for intervention programs.

Small cell carcinoma of cervix (SCCC) is a rare disease with highly aggressive behaviour and is pathologically hard to diagnose. In this study, the clinicopathological features, diagnosis, treatment and prognosis of the condition were examined. Clinical records and follow-up data of 7 cases of SCCC were retrospectively studied. Our results showed that five non-recurrent cases initially presented irregular vaginal bleeding or increased apocenosis of varying degrees. Pathological examination revealed that the stroma was diffusely infiltrated with small monomorphous cells ranging from round to oval shape. Three cases were immunohistochemically confirmed. One case was accompanied with squamous cell cancer. Of the 7 cases, one case was classified as stage I b1, two stage I b2, one stage IIa, one stage IIb, and one stage IIIb. On the basis of their stages of condition, one subject with stage III b underwent chemotherapy, and one with stage Ib2 received extensive hysterectomy plus pelvic lymphadenectomy, while the other 5 cases were treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy. Of the 7 patients, 4 had relapse-free survival of 14, 14, 16 and 28 months respectively. It is concluded that SCCC is an aggressive tumor with propensity for early pelvis lymph node metastases. Early-stage patients should be treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy.

OBJECTIVETo understand the incidence of colorectal cancer in population drinking or not and to validate the relationship between drinking and colorectal cancer.

METHODSThe data obtained from a questionnaire used in a population-based prospective screenings study in ten countries of Jiashan County was examined. A total of 64,102 men and women aged 30 y and older without history of cancer at baseline and a subcohort of 29,044 of them drinking past and current was conducted. Cox regression model was applied to estimate relative risk (RR).

RESULTSAfter 10 years follow-up,107 colon cancer and 135 rectal cancer cases were identified. Among drinkers and abstainers, the incidence density of colorectal cancer was 36.18 per 100 thousand and 37.26 per 100 thousand, respectively and there wasn't statistical significance(Z=0.52, P>0.05); The crude RR (95%CI) for drinker compared with never drinkers was 0.97(0.75 approximately 1.25), and the multivariable-adjusted RR (95%CI) was 1.13(0.87 approximately 1.48). The research power of this study was 96.99%.

CONCLUSIONAlcohol drinking isn't one of the risk factors of colorectal cancer among Jiashan County population.

Adult ; Alcohol Drinking ; adverse effects ; China ; epidemiology ; Cohort Studies ; Colorectal Neoplasms ; epidemiology ; etiology ; Female ; Follow-Up Studies ; Humans ; Incidence ; Male ; Proportional Hazards Models ; Prospective Studies ; Risk Factors ; Surveys and Questionnaires

OBJECTIVE: To investigate the role of adult cardiomyocytes in the differentiation of mesenchymal stem cells (MSCs) into cardiomyocytes. METHODS: Rat MSCs were isolated by a Percoll's gradient solution and cultured in low-glucose Dulbecco' s modified Eagle' s medium (DMEM). After 2 passages, cell-surface antigen CD34, CD71 and CD90 for rat MSCs were determined by flow cytometry, and these MSCs were transfected with pEGFP-N3 by Lipofectamine2000. Then those MSCs labeled with GFP, were cultured in contacted, nocontacted and conditioned with adult rat myocardiocytes. Immunofluorescence staining against alpha-actin, desmin, and troponin-T were performed after 1 week. RESULTS: Immunofluorescence staining was positive against alpha-actin, desmin, and troponin-T on MSCs in contacted culture group. In contrast, no alpha-actin, desmin, and troponin-T expression on MSCs were observed in the noncontacted culture group and the conditioned culture group. CONCLUSION Direct cell-to-cell contact between MSCs and adult cardiomyocytes may induce differentiation of MSCs into cardiomyocytes.
Animals ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; Cell Communication ; Cell Differentiation ; physiology ; Cell Separation ; Cells, Cultured ; Coculture Techniques ; Female ; Male ; Mesenchymal Stem Cells ; cytology ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Sprague-Dawley ; Thy-1 Antigens ; analysis

The aim of this study was to investigate the expression of transferrin receptor 2 (TfR2) mRNA in bone marrow mononuclear cells (BMMNC) of children with hyperplastic anemia (HA), to analyze the correlation of TfR2 mRNA expression level with Hb level, bone marrow erythroid hyperplasia, iron status in body and underlying diseases, and to evaluate the role of TfR2 in erythroid hemopoiesis and the useful value in diagnosis of HA. The experiment was divided into 2 groups: test group, in which 40 patients with HA were enrolled, and control group in which 10 patients without erythroid disorders and hematological malignancies confirmed by bone marrow examination were enrolled. The bone marrow samples of patients in mentioned above 2 groups were collected, the TfR2 mRNA expression in BMMNC of patients with HA was detected by fluorescence-quantitative PCR, the correlation of HA with bone marrow erythroid hyperplasia, iron status of body and underlying diseases was analyzed. The results showed that the relative level of TfR2 mRNA expression in HA patients was significantly higher than that in control patients. The TfR2 mRNA expression level negatively correlated with Hb level in peripheral blood (r = -0.715), while it positively correlated with ratio of bone marrow erythroblasts (r = 0.533). It is concluded that TfR2 mRNA expression in HA patients increases and closely correlates with hyperplasia status of bone marrow and anemia level in peripheral blood.
Adolescent ; Anemia ; metabolism ; pathology ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Erythroid Precursor Cells ; metabolism ; Female ; Humans ; Infant ; Male ; RNA, Messenger ; metabolism ; Receptors, Transferrin ; metabolism