To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.

OBJECTIVETo assess the population genetic diversity and genetic structure and screen species-specific bands for identification of Changium smyrnioides and Chuanminshen violaceum.

METHODSeven wild populations of Changium smyrnioides and one cultivated population of Chuanminshen violaceum were studied by ISSR analysis. The population genetic diversity and population genetic structure were assessed by using POPGENE software.

RESULTA total of 152 ISSR markers were scored, among which 136 (90.8%) were polymorphic. The values of Gst tended to be high (mean Gst = 0.575). The level of genetic divesity of Changium smyrnioides (A = 1.272; P = 27.26%; I = 0.132; H = 0.087) was higher than that of Chuanminshen violaceum (A = 1.217; P = 21.7; I = 0.103; H = 0.067).

CONCLUSIONThe genetic variation of Changium smyrnioides is high and the majority of genetic variation occur among populations. Substantial genetic divergence is shown by cluster analysis (UPGMA) to befound between Changium smyrnioides and Chuanminshen violaceum at DNA level. In addition, one species-specific marker has been obtained in Chuanminshen violaceum. The phylogenetic relationship of two species has also been discussed.

Apiaceae ; classification ; genetics ; China ; Cluster Analysis ; DNA, Plant ; genetics ; Ecosystem ; Gene Frequency ; Genetic Markers ; Genetic Structures ; Phylogeny ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Repetitive Sequences, Nucleic Acid ; Species Specificity

Objective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.

Objective] To amplify and sequence the light chain of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum. [Methods] By comparing the conserved regions at each end of the nucleotide sequences of murine germ line genes enco ding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of V L gene. The hybridoma cells secreting anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger′s method. The V L gene was compared with GenBank and published mouse V L genes. [Results] The full length of V L gene was 318 bp. The V L gene was a member of mouse Ig ? light chain subgroup IV and generated from rearrangement of germ line V and J? 4 genes. The V L gene sequence has been registered by GenBank(accession No. AF206720). [Conclusion] The obtained V L gene was a potentially functional gene of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum .

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) efficiently in pilot scale, cells of rhNDPK-A producing E. coli were homogenized by high pressure under 4 degrees C, 950 Pa. The insoluble debris was removed by microfiltration and the soluble portion was concentrated by ultrafiltration. The resulted crude sample was loaded on DEAE-sepharose Fast Flow. The target fraction was collected and then load on Cibacron Blue 3GA Sepharose CL-4B. Eluted with buffer containing ATP from the AC column, rhNDPK-A was polished with ultrafiltration. The results showed that after homogenized 2 rounds, 1500g cells of E. coli brought crude sample containing 47.6g NDPK-A. Treated with microfiltration and ultrafiltration, 27.3g of NDPK-A were recovered from this bacteria homogenate. After 2-step purification with column chromatography and then polished with ultrafiltration, 17.2 g rhNDPK-A were collected with purity of 96.3%. The recovery of the whole purification process was 36.2%, and the productivity of rhNDPK-A was 1.15 g per 100 g wet cells. Comparing the recovery of each purification step, it was found that the recovery of polish is higher than that of affinity chromatography, which is higher than that of ion exchange chromatography. The limit step was the process of sample pretreatment among the 4 purification steps. Combine with the fermentation results reported before, it was deduced that the productivity of rhNDPK-A was 510 mg/L. In conclusion, an easily controlled purification condition with high yield provides material for the translation researches of NDPK; In addition, it was suggested the crucial step determine the recovery of non-secretive recombinant proteins might be the process of sample pretreatment, not be the process of column chromatography.
Chromatography, Affinity ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Humans ; NM23 Nucleoside Diphosphate Kinases ; genetics ; metabolism ; Pilot Projects ; Recombinant Proteins ; isolation & purification ; metabolism ; Ultrafiltration

OBJECTIVETo identify the epidemical characteristics of suicidal tendency among middle-school students in cities of China and to explore the main factors leading to suicidal tendency in adolescents.

METHODSMulti-stage cluster sampling method was used to select 9015 students in grades 1, 2, 3 and 4 respectively from 25 general middle schools in Beijing, Hangzhou, Wuhan and Urumqi of China in June 2006 and field investigation was carried out through "China Global School-based Student Health Survey (GSHS) Questionnaire".

RESULTSAmong the students in the four cities, the incidence rates of suicidal ideation were from 14.4% to 20.8% with an average of 17.4%. The incidence rates of suicidal plan were from 6.8% to 9.7% with an average of 8.2% and were different among cities. 15.0% of the boys had suicidal ideation and 6.7% of them made a suicidal plan comparing to 19.7% of girls having had suicidal ideation and 9.5% of them made a suicidal plan. The two kinds of suicidal tendency in girls were all higher than those in boys. City, age, gender, grade, days and type of being bullied, depression, close friends and having received health education on coping with stresses were factors influencing suicidal tendency of students. Days of being bullied and suicidal tendency showed a dose-response relation.

CONCLUSIONSuicidal tendency seemed common in middle-school students. Training on 'coping the issue' should be strengthened and harmonious environment should be improved in middle-schools.

Adolescent ; China ; epidemiology ; Data Collection ; Female ; Humans ; Incidence ; Male ; Students ; psychology ; Suicide ; psychology ; statistics & numerical data ; Urban Population

OBJECTIVETo study the clinicopathological and genetic features of desmoid-type fibromatosis, and to investigate the feasibility of detecting trisomy 8 in formalin fixed, paraffin embedded (FFPE) tissue by fluorescence in-situ hybridization (FISH).

METHODSA total of 96 cases were included in this study. All patients had clinical information. Histopathologic and immunohistochemical evaluations were available in 69 cases, and ultrastructural evaluation was done in 2 cases of desmoid-type fibromatosis. FFPE tissue sections were available in 20 tumors for the trisomy 8 detection by FISH.

RESULTSThere were 20 male and 76 female patients with ages ranging from 8 to 86 years (mean 35.3 years). Clinically, there were 44 extra-abdominal tumors, 28 abdominal wall tumors and 23 intra-abdominal lesions mostly involving the mesentery. Most cases presented with nodular or funicular masses with white firm cut surfaces, measuring 0.6 to 24.0 cm (mean 8.4 cm) in size. Histologically, desmoid-type fibromatoses showed longitudinal fascicles of spindle fibroblasts and myofibroblasts in a predominantly collagenous background. The tumor cells stained positive for vimentin, alpha-smooth muscle actin, desmin, and beta-catenin (47.8%, 33/69). Ultrastructurally, most tumor cells had features of fibroblasts, including rich endoplasmic reticulum and Golgi apparatus. Some tumor cells were myofibroblast-like cells exhibiting intercellular junctions, fibronexous junctions and stress fibers. Trisomy 8 was detected in 6 of 20 cases of desmoid-type fibromatosis including 5 of the 8 recurrent tumors but only one of the 12 primary tumors. The latter tumor also recurred three years later.

CONCLUSIONSDesmoid-type fibromatosis is an intermediate (locally aggressive) tumor that occurs predominantly in young females. The lesion consists of fibroblasts and myofibroblasts with the latter showing characteristic features including stress fibers and fibronexous junctions. Trisomy 8 can be detected in FFPE tissue by FISH, and its presence serves as a useful predictor of tumor recurrence and may define a subtype of desmoid-type fibromatosis with high recurrence rate.

Actins ; analysis ; Adult ; Aged ; Aged, 80 and over ; Child ; Chromosomes, Human, Pair 8 ; genetics ; Desmin ; analysis ; Feasibility Studies ; Female ; Fibromatosis, Abdominal ; genetics ; metabolism ; pathology ; Fibromatosis, Aggressive ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Male ; Mesentery ; Middle Aged ; Muscle, Smooth ; chemistry ; Neoplasm Recurrence, Local ; Peritoneal Neoplasms ; genetics ; metabolism ; pathology ; Trisomy ; Vimentin ; metabolism ; beta Catenin ; analysis

Acrylic Resins ; adverse effects ; Adult ; Breast Implantation ; adverse effects ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Middle Aged

AIMTo observe possible mechanism that endurance training can enhance anti-fatigue capability, and that blood redistribution by analyzing some biochemical indexes of endurance-trained mice after exhaustive exercise.

METHODSThe model was set up by exhaustive exercise. The indexes include the activity of SOD, CAT and POD and the MDA content in serum and the NO content in liver, muscle, heart and serum.

RESULTSAfter exhaustive exercise, the SOD activity in serum and the NO content in liver significantly decrease (P < 0.05 - 0.01), and the activity of POD and CAT, the NO content in serum and muscle significantly increase (P < 0.05 - 0.01), but the rest insignificantly change in non-endurance (P > 0.05). In endurance group, the CAT activity in serum are significantly higher than in non-endurance (P < 0.05), and the NO content in serum is significantly lower than in non-endurance (P < 0.01), but the rest are insignificantly different between two groups (P > 0.05). After 24h restoration, in non-endurance group, the CAT activity and the MDA content in serum and the NO content in liver significantly rise (P < 0.05-0.01), and the NO content in muscle and serum significantly decrease (P < 0.05), but the rest insignificantly change (P > 0.05). In endurance group, the SOD activity in serum and the NO content in liver, serum and heart significantly rise (P < 0.05), and the CAT activity in serum significantly decreases (P < 0.05), but the rest insignificantly change (P > 0.05). In endurance group, the CAT activity and the MDA content in serum are significantly lower than in non-endurance (P < 0.05), but the NO content in heart is higher than in non-endurance (P < 0.05). The rest are insignificantly different between two groups (P > 0.05).

CONCLUSIONThe possible mechanism, which endurance training can enhance anti-fatigue capability, is relative to enhance the capability to resume balance. Blood redistribution are possibly relative to change to the NO content.

Animals ; Catalase ; blood ; Liver ; chemistry ; Male ; Malondialdehyde ; blood ; Mice ; Muscles ; chemistry ; Nitric Oxide ; analysis ; blood ; Peroxidase ; blood ; Physical Conditioning, Animal ; physiology ; Physical Endurance ; physiology ; Superoxide Dismutase ; blood

OBJECTIVETo investigate the effects of atrial natriuretic peptide (ANP) on ischemia and reperfusion cochlea in guinea pigs.

METHODSThe guinea pigs were randomly allocated into four groups: experiment groups (A1 and B1) and control groups (A2 and B2). Cochlear ischemia and reperfusion was induced by thrombus and thrombolysis method. In experiment group A1, ANP was administered 10 min before the ischemic insult. In experiment group B1, ANP was administered at the beginning of reperfusion. In control groups, instead of ANP, normal sodium was injected. The blood flow of cochlea (CoBF) was monitored continuously with laser Doppler flow meter and the threshold of auditory brainstem response (ABR) was measured.

RESULTSBefore the induction of ischemia, the CoBF of experiment group A1 was higher than that of the control group A2. From the reperfusion moment to the end of the experiment, there was no difference between the CoBF of the two groups. In B1 and B2 groups, no difference could be seen between the two groups before the induction of ischemia. After reperfusion, the blood flow of control group B2 recovered to 70% of the base level, while the CoBF of experiment group B1 restored to almost the same level of the beginning. Before ischemia, the ABR threshold of the four groups had no difference. At 30 min of ischemia, the threshold of experiment group Al was lower than that of control group A2. And there was no difference in experiment group B1 and control group B2. At 30 min and 60 min of reperfusion, the threshold of experiment group B1 was significantly lower than that of control group B2. No difference could be seen between experiment group A1 and control group A2.

CONCLUSIONSAdministration of ANP at the beginning of reperfusion protects the cochlea from ischemia and reperfusion injury. The administration can not only increase the CoBF, but lower the ABR threshold.

Animals ; Atrial Natriuretic Factor ; pharmacology ; Cochlea ; blood supply ; drug effects ; physiopathology ; Disease Models, Animal ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Reperfusion Injury ; drug therapy ; physiopathology