Objective To identify the polyreactivity of a monoclonal natural anti-keratin autoantibody 3B4 and to analyze its possible molecular me chanism.Methods enzyme-linked immunosorbent assay (ELISA)and immunohistoche mistry were applied to test the binding reactivity of 3B4 against different anti gens and tissues.The variable region genes and their amino acid composition wer e sequenced.Results 3B4 could reacted with a range of antigens and tissues,i n addition to keratin and skin.The variable region genes of its light chain and heavy chain showed high homology with germline genes VK1 am4 and VH1 J558.42.H CDR3 region,which mainly composed of short side chain amino acids(from 294 to 324 nucleotides around the heavy chain),was the only motif that differs from ot her highly homologous immunoglobulin genes.Conclusions The monoclonal natural anti-keratin autoantibody 3B4,with its variable region genes highly homologo us to germline genes,is highly polyreactive.The flexibility of HCDR3 may contr ibute to the polyreactivity.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

0.05).At the late stage (8-48 h) of incubation,the presence time of E7_(49-57)/K~b was significantly pro- longed on the surface of Tat-E7_(49-57)-incubated cells than that on the surface of other peptides-incubated cells (all P

Objective To provide supporting evidence to prevention and controlling of caries in preschool children in Huangpu District(Eastern Part), by studying the influence of their habits at home on the prevalence of caries. Methods The oral health of preschool children in Huangpu District was examined, and their habits at home were recorded through survey questionnaire, including sugary diet, tooth brushing, parents′ attitude for oral health. Results Sugary diet could affect the prevalence of caries.There was no association between dental caries and times of tooth brushing.Parents′ attitude for oral health is the risk factor for the children′s caries. Conclusion Because of the higher prevalence of caries in the children in our district, effective measures should be taken.In addition, policy should consider how to ensure that children with high-risk dental caries receive adequate prevention and early care.

Objective To investigate the effect of mild hypothermia on brain edema after severe head injury (SHI) and its clinical significance. Methods Thirty-six patients with SHI were randomly divided into conventional therapy group (control group) and mild hypothermia therapeutic group (therapeutic group), with 18 patients in each group. The patients in the therapeutic group were treated with ice-cap, ice-blanket and ice-bag and lytic cocktail Ⅰas adjuvants, and a rectal temperature of 33 ℃ was maintained for 3~5 d. Intracranial pressure (ICP) of all the patients after operation were monitored postoperatively. The size of brain edema was measured with CT on day 0, 1, 3, 7, 14, 21 respectively after SHI. The size of brain edema at the trauma cite and ICP as well as the clinical outcome were compared and analysed. Results ICP of the control and therapeutic groups were（2.87±0.26）kPa and (1.67±0.23) kPa respectively on day 3 (P<0.01), and were (3.15±0.24)kPa and (1.78±0.24) kPa respectively on day 7 after brain injury (P<0.01). The area of brain edema was the largest on day 14 in the control group (140.90±22.95 cm3)and on day 3 in the therapeutic group (95.83±14.97 cm3). At day 14 posttrauma, the edema area in therapeutic group was 42% smaller than that in the control group (P<0.05). The rates of consciousness recovery within a week were 22.2% (4/18) in the control group and 55.6% (10/18) in the therapeutic group (P<0.05). The rates of unfavorable prognosis was 61.1% (11/18) in the control group and 33.3% (6/18) in the therapeutic group. Conclusion Mild hypothermia treatment can improve the prognosis through alleviating brain edema and stabilizing ICP after SHI.

Objective To investigate the effect of mild hypothermia on brain edema after severe head injury (SHI) and its clinical significance. Methods Thirty-six patients with SHI were randomly divided into conventional therapy group (control group) and mild hypothermia therapeutic group (therapeutic group), with 18 patients in each group. The patients in the therapeutic group were treated with ice-cap, ice-blanket and ice-bag and lytic cocktail Ⅰas adjuvants, and a rectal temperature of 33 ℃ was maintained for 3~5 d. Intracranial pressure (ICP) of all the patients after operation were monitored postoperatively. The size of brain edema was measured with CT on day 0, 1, 3, 7, 14, 21 respectively after SHI. The size of brain edema at the trauma cite and ICP as well as the clinical outcome were compared and analysed. Results ICP of the control and therapeutic groups were（2.87±0.26）kPa and (1.67±0.23) kPa respectively on day 3 (P<0.01), and were (3.15±0.24)kPa and (1.78±0.24) kPa respectively on day 7 after brain injury (P<0.01). The area of brain edema was the largest on day 14 in the control group (140.90±22.95 cm3)and on day 3 in the therapeutic group (95.83±14.97 cm3). At day 14 posttrauma, the edema area in therapeutic group was 42% smaller than that in the control group (P<0.05). The rates of consciousness recovery within a week were 22.2% (4/18) in the control group and 55.6% (10/18) in the therapeutic group (P<0.05). The rates of unfavorable prognosis was 61.1% (11/18) in the control group and 33.3% (6/18) in the therapeutic group. Conclusion Mild hypothermia treatment can improve the prognosis through alleviating brain edema and stabilizing ICP after SHI.

OBJECTIVETo study the mechanism of learning and memory dysfuction in the transgenic mouse expressing human tau 40 isoform with P301L mutation (F10).

METHODSThe human tau protein expression and phosphor-tau protein levels were detected with Western blot method. The neurofibrillary tangles were observed with Bielshowsky silver stain. The behavior changes of learning and memory were observed by open field test and passive avoidance test. Acetyleholine level, activities of acetycholinesterase and choline acetyltransferase of whole brain was detected by colorimetry method. The nitric oxide level of whole brain was detected by nitrate enzyme reduction method.

RESULTSExogenous human tau gene was expressed and an elevation of phosphor-tau protein level in 7 and 3-month transgenic mice's hippocampus andcerebrocortex was observed. The neurofibrillary tangles were observed in cerebrocortex of 7-month transgenic mice; the 7-month transgenic mice also presented an evident reduction of learning and memory ability and nitric oxide level of the whole brain, but not changes in acetylcholine level, acetycholinesterase activity, choline acetyltransferase activity and expression in whole brain.

CONCLUSIONTau transgenic mice (F10) can still inherit their parents' biologiccal characters, and develop learning and memory dysfunction awnodh san obvious decrease in nitric oxide level of whole brain in the 7-month old mice, suggesting a decrease of nitric oxide level of whole brain would be involved in the mechanism of learning and memory dysfunction in these transgenic mice.

Acetylcholine ; metabolism ; Acetylcholinesterase ; metabolism ; Animals ; Brain ; physiopathology ; Choline O-Acetyltransferase ; metabolism ; Humans ; Membrane Proteins ; genetics ; Memory Disorders ; genetics ; physiopathology ; Mice ; Mice, Transgenic ; Mutation ; Nitric Oxide ; metabolism

Objective To investigate the association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms with metabolic syndrome among Kazakh and Han ethnicities in Xinjiang.Methods PCR-RFLP was used to detect 802 subjects' lipoprotein lipase Hind Ⅲ and S447Xgenotypes (including 201 controls and 200 metabolic syndrome patients in Kazakh and Han ethnicities, respectively). Results (1) Frequencies of H + H-/H-H- genotype (32.50% vs.47.76%), H- allele( 18.00% vs. 28.86%), SX/XX genotype (8.00% vs. 22.39%) and X allele (4.00%vs. 12.44% ) for metabolic syndrome in Hah ethnicity were all significantly lower than those in controls (P＜ 0.01 ). (2) The frequencies of H + H-/H-H- genotype (33.50% vs. 46.80% ), H- allele (22.00% vs. 28.60%), SX/XX genotype (10.50% vs. 22.90%) and X allele (5.50% vs. 12.44% ) in patients with metabolic syndrome in Kazakh were all significantly lower than those for controls (P＜0.01). (3) The frequencies of lipoprotein lipase gene Hind Ⅲ and S447X genotypes and alleles in Kazakh were not significantly different from Han (all P＞0.05). (4)The levels of waist circumference, systolic blood pressure, diastolic blood pressure, triglyceride and FPG in H + H-/H-H- and SX/XX genotype were significantly lower than those in H + H + and SS genotype.HDL-C was significantly higher than that in H + H + and SS genotype (P＜0.05). (5) The frequencies of H + H + and SS genotype increased along with the increase in number of metabolic syndrome component. Conclusion The lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms were associated with metabolic syndrome risk in Kazakh, and H + H-/H-H- genotype, H- allele, SX/XX genotype and X allele might have served as protective factors of metabolic syndrome. H + H-/H-H- and SX/XX genotype seemed to have had beneficial effects for all the metabolic syndrome components, and the frequencies of H + H + and SS genotype were increasing along with the increase of number in the metabolic syndrome components.

Objective To compare the immunogenicity of rAAV2/1 and rAd5 expressing HIV-1 gag in BALB/c mice.Methods BALB/c mice wcre immunized with rAAV2/1-gag or rAd5-gag once or twice.HIV-1 specifc cellular immune responses were analyzed by in vivo CTL and intracellular cymkine staining assays.HIV-1 Gag specifc antibodies were tested by ELISA.Results Mice immunized with rAd5-gag once induced stronger Gag specific cellular immune responses and similar level of Gag specific antibody compared with rAAV2/1-gag.Mice immunized with rAd5-gag reached the peak immune responses more rapidly than rAAV2/1-gag.However,mice immunized with rAAV2/1-gag twice elicited better Gag specific IgG.Conclusion rAd5-gag induced strong HIV-1 specifc cellular and antibody responses,and rAAV2/1-gag induced high level of HIV-1 specific IgG and moderate cellular immune responses.

OBJECTIVETo extract two kinds of phenols 4-hydroxy-3, 5-dimethoxy-4-(2-oxopropyl) cyclohexa-2, 5-dien-l-one and 6-methoxy-5,7-dihydroxy coumarin (named as I and H compounds respectively) from Ajania salicifolia and to investigate their antioxidation and cytotoxicity to tumors and explore their pro-apoptosis mechanism.

METHODSThe antioxidant activities of two compounds were assessed by ABTS and DPPH radical-scavenging assays. Two compounds were evaluated for their cytotoxicity against human chronic myelogenous leukemia (K562) cells using the MIT assay. The expression of NF-kappaB P65 mRNA in K562 apoptotic cells was measured by reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative PCR. In addition, protein expression levels of the NF-ICB P65, p-Akt, Fas, P-catenina and E-cadherin were also measured by Western blot.

RESULTS(1) We found that compound I displayed significant inoxidizability, while compound II had no obvious antioxidizability. (2) In cytotoxicity experiments, compound I didn't display cytotoxicity while compound H displayed obvious cytotoxicity. (3) Compared with the blank group, the expression of NF-kappaB P65 mRNA in K562 cell after treatment with compound II was obviously up-regulated. (4) Compared with the blank group, the expression levels of NF-kappaB P65, Fas, beta-catenina and E-cadherin were significantly increased in compound II treated groups and it appeared obvious dose-effect relationship between the expression of protein and drug concentration.

CONCLUSIONTwo phenols have obvious antioxidizability and cytotoxicity respectively. On the one hand, the tumor-suppressing mechanism of compound II maybe act by up-regulation the expression of NF-kappaB P65 and Fas protein; thereby, affecting the classical Fas apoptosis signaling pathways. On the other hand, it can also up-regulate the expression of protein beta-catenin and E-cadherin, which participate in the adhesion between cells, and accordingly, playing an important role in preventing the proliferation and metastasis of cancer cells.

Apoptosis ; Asteraceae ; chemistry ; Cadherins ; metabolism ; Humans ; K562 Cells ; Oncogene Protein v-akt ; metabolism ; Phenols ; chemistry ; Signal Transduction ; Transcription Factor RelA ; metabolism ; Up-Regulation ; beta Catenin ; metabolism ; fas Receptor ; metabolism