Objective To investigate the early comprehensive intervention on neonatal hypoxic ischemic encephalopathy (HIE) effects and the biological mechanisms.Methods 120 cases of HIE were randomly divided into intervention group and control group.Under the basis of drug treatment,the intervention group were given the different perceptual stimuli,audiovisual stimulation,active and passive limb training early comprehensive intervention,the control group only to normal drug treatment.The mental development index(MDI) and psychomotor develo pmental index (PDI) were applied after 3,6,12,and 18 months.S100B protein levels were detected using enzyme immunoassay.Results MDI score in the intervention group with HIE,from the beginning of the 12 month,was higher than that in the control group,there was significant differences (12th month:(96.6±9.4)vs (86.8±9.5),P<0.01).PDI score from the beginning of the 6 month,was higher in the intervention group with HIE than that in the control group,there were significant differences (6th month:(91.6±9.2)vs (82.2±11.0),P<0.01).Serum S100B protein level has a tendency to fall for all HIE patients with the old growth.Born in two groups of serum S100B protein levels were higher in neonate period,but there was no difference(P>0.05).From the beginning of the 3rd month,serum S100B protein levels in the intervention group were reduced significandy,compared with the control group at each time point level(3rd month:(1.29±0.15)ng/L vs(1.40±0.14)ns/L;6th month:(1.06±0.21)ng/L vs(1.26±0.22)ng/L,P<0.01).Conclusion Early intervention can improve the mental and motor development of children with HIE,and serum S100B protein is involved in the mechanisms.

Objective To realize the construction and expression of human dsFv genes to rabies virus. Methods The genes of dsFv VH and VL were amplified by PCR-based point mutagenesis strategy. Then the genes were cloned into plasmid pET-22b (+). E. coli BL21(DE3) was transformed with the recombinant expression plasmid and the protein was induced in Luris-Bertani medium by addition of IPTG. The inclusion body proteins of VH and VL were denaturalized by GuHCl, and then re-naturalized in refolding solution to form dsFv fragments. Afterwards, we evaluated the antigen-binding activity of the dsFv and its stability as compared with its originative scFv. Results The fragment genes of dsFv to rabies virus were constructed successfully by PCR-based point mutagenesis strategy. Sequence analysis proved that cysteines were introduced into the position 44 aa of VH and position 100 aa of VL. The dsFv VH and VL genes were expressed in PET22b(+)/BL21(DE3). The VH and VL protein folded into the active dsFv antibody fragments which showed specific binding capability to rabies virus. Conclusion We succeeded in achieving the stabilization of the human scFv to rabies virus and obtaining the active human dsFv antibody fragments. This established a solid basis for further study of the biological activaty and clinical application of dsFv to rabies virus.

To change the medicine teaching model and cultivate the medicine students to have Tongji feature ( good backgrounds,valuing practices,knowing scientific research,examining patients ),our teaching and research section takes PBL method in theory and clinical teaching.During 5 years,we have built PBL teaching system of oncology successfully,including the high quality of PBL teaching plans and the PBL teachers who have rich experience and innovative thinking,and the object evaluation system of PBL teaching.It is a constructive action of PBL teaching method for reform of teaching in oncology.PBL teaching method in oncology will be consummated persistently.

BACKGROUND: Treatment of ulcerative colitis (UC) with herbs-partition moxibustion is of good intervenient effect, whether the effect is related to neutrophil apoptosis in the pathogenic process of UC?OBJECTIVE: To investigate the changes of neutrophil appptosis rate of rats with UC and the regulative effect of medicinal cake moxibustion. DESIGN: A randomized controlled trial based on SD rats.SETTING: Shanghai Research Institute of Acupuncture and Meridian.MATERIALS: The experiment was completed from August 2002 to June 2003. A total of 30 healthy male SD rats of clean grade, weighing (140±20) g,provided by Experimental Animal Center of Shanghai University of Traditional Chinese Medicine, were at random divided into 2 groups: model building group (n=20) and normal control group (n=10).METHODS: Rat UC models were set up with immunological method. After the models were successively set up, four model rats and two normal rats were randomly selected for pathological observation of distal colon tissue. The rest models were divided as model group and herbs-partition moxibustion group with 8 rats for each, and the rest 8 normal rats were taken as control group. The rats of herbs-partition moxibustion group were given herbs-partition moxibustion at tian shu (ST-25) of both sides and qi hai (CV-6) points; the herbs cones was made from Radix Aeoniti Praeparata,Cortex Cinnamoni, and so on. About 90 mg moxa cone was put on the herbs cones for moxibustion, once a day, for 10 times. After treatment, all rats of the three group were executed, then the peripheral monocytes were isolated for cell culture. A 1:50 diluted supernatants of each group were taken, and the same volume of RPMI-1640 culture fluid was used as blank control group. They were respectively incubated together with neutrophils of peripheral blood of normal rats. The neutrophil apoptosis rates were measured by using flow cytometry.MAIN OUTCOME MEASURES: Comparison of neutrophil apoptosis rates of rats in each group.RESULTS: Totally 20 rats in model group and 10 in normal control group were accepted. Then 4 rats in model group and 2 in normal group were sacrificed and observed with tissue pathology, and 16 and 8 rats entered the final analysis in model and normal control groups respectivelly. ① The neutrophil apoptosis rate of UC rats was significantly lower that that of rats in blank control group [(16.34±2.80) ％, (52.33±9.94) ％, q= -35.99, P＜ 0.01]; but there was no significant diffenence between normal control group [(48.79±11.33) ％] and blank control group, (P＞0.05). ② After treatment the neutrophil apoptosis rate of herbs-partition moxibustion group [(36.03±8.31) ％] was significantly higher than that of UC model group (q=19.69, P ＜ 0.01), but it was still lower than that of normal control group (q= -16.30, P ＜ 0.05).CONCLUSION: One of the main mechanism for herbs-partition moxibustion to treat UC might be regulating and/or reducing the inhibited state of neutrophil apoptosis in UC.

Objective To investigate the effects of magnesium isoglycyrrhizinate on liver regeneration and hepatic function following partial hepatectomy in thioacetamide induced cirrhotic rats.MethodsForty-five cirrhotic Wistar rats undergoing 2/3 hepatectomy were randomly assigned to control group ( Group A), Group B and Group C starting the day of hepatectomy, rats in Group B were injected 60mg per kg body-weight magnesium isoglycyrrhizina daily intraperitoneally until the day of sacrifice. Rats in Group A recevied same dose of sodium chloride. In Group C, magnesium isoglycyrrhizina was administered daily 3 days before hepatectomy until the rats were sacrificed. Liver function, serum HGF, serum PLA2,BrdU labelling index and percentage of intial liver weight on days 1, 2 and 7 post hepatectomy were assessed. ResultsRats in Group A had significantly lower BrdU labelling index and serum HGF level than Group C ( t = 2. 831, P ＜ 0.05; t = 3.427, P ＜ 0.05 ) and a markedly higher level of serum PLA2 than the other groups on day 1 posthepatectomy ( Group B t = 2. 794, P ＜ 0.05; Group C t = 2. 902, P ＜ 0.05 ).Rats in Group A had a lower BrdU labelling index and a more increased level of serum PLA2 than Group B and Group C on day 2 posthepatectomy ( BrdU labelling index: Group B t = 2. 736, P ＜ 0.05; Group C t =3.083, P＜0.05; PLA2: Group B t =2.794, P＜0.05; Group C t =2.902, P＜0.05), but had no significant difference in HGF level with the other two groups. The three groups were similar in ALT, AST,TP and intial liver weight on days 1,2 after operation. On the 7th day posthepatectomy, rats in Group A had a higher level of AST ( Group B t = 4. 508, P ＜ 0.05; Group C t = 2. 967, P ＜ 0.05 ) and a lower level of TP ( Group B t = 2. 838, P ＜ 0.05; Group C t = 2. 743, P ＜ 0.05 ), lower liver weight than the other two groups ( Group B t = 3.316, P ＜ 0.05; Group C t = 4. 093, P ＜ 0.05) but there was no difference between the three groups in BrdU labelling index, HGF and PLA2 level. Rats in Group C had significantly higher BrdU labelling index and serum HGF level than the rats in Group B on day 1 after hepatectomy( t = 2. 831, P ＜0.05; t = 2. 836, P ＜ 0.05 ).ConclusionsMagnesium isoglycyrrhizinate inhibits aminopherase release and enhancing liver regeneration in cirrhotic rats after partial hepatectomy.

Objective To detect steroidogenic acute regulatory protein (StAR) expression in apolipoprotein E-deficient mice at different ages and serum lipid levels. Methods Nighty-six C57BL/6J and apoE-/- mice were enrolled, which were divided into 16 groups with 6 mice per group according to age (1 day, 1, 3, 5 months), sex and genotype (C57BL/6J and apoE-/-). The serum lipid levels in C57BL/6J and apoE-/- mice were detected by commercial kits. StAR mRNA and protein expressions in liver were detected by Real-time PCR and Western blot respectively. Results ApoE-/- mice had higher LDL-cholesterol and lower HDL-cholesterol compared with C57BL/6J mice of the same age and sex. StAR mRNA and protein expressions were decreased following with aging in C57BL/6J mice. However, in apoE-/- mice with higher lipid levels, StAR mRNA and protein expressions were changed with the lipid levels other than ages. StAR mRNA and protein increased in the early stage, and then decreased with the increasement of lipids levels. Conclusions StAR could affect lipids levels and may be an effective regulator for atherosclerosis and other cardiovascular diseases.

Objective: To investigate the inhibitory effect of survivin antisense oligonucleotide (ASODN) on the expression of survivin gene and proliferation of human hepatocellular carcinoma cell line SMMC-7721. Methods: The 20 mer antisense oligonucleotide (ASODN) targeted to the promoter region of survivin mRNA was designed and synthesized. The expression of survivin gene in hepatocellular carcinoma cell line SMMC-7721 was blocked by means of ASODN transfection mediated by DOTAP liposomal reagent. The changes of survivin protein and mRNA expression after transfection were as-sessd by Western blot and in situ hybridization, respectively. The apoptotic rate was detected by flow cytometer. The changes of cell adherent rate, cell growth activity, and the inhibitory rate of cell growth were also studied. Results: The expression of survivin protein and mRNA were decreased markedly after survivin ASODN transfection. Meanwhile, the cell adherent rate also decreased markedly while the apoptotic rate increased markedly. Conclusions: Transfection of ASODN targeted to the promotor region of survivin mRNA by DOTAP liposomal transfection reagent could down-regulated the expression of survivin protein and mRNA significantly in 7721 cell line and inhibit the proliferation of cancer cells. Survivin could be an important target in the therapy of hepatocellular carcinoma.

Objective To explore the clinical value of multi-slice CT enhancement perfusion imaging (MSCTPI) in diagnosis and differential diagnosis of thyroid disease. Methods Thirty-three patients with benign thyroid diseases were enrolled in the benign group, and were divided into subgroups of nodular goiter (n=17) and thyroid adenoma (n=16), while 10 patients with thyroid carcinoma were enrolled in the malignant group. All patients underwent routine CT scanning and MSCTPI with GE LightSpeed 16-detector row CT scanner. Time-density curve (TDC) of common carotid for benign thyroid diseases and thyroid carcinoma was depicted. Perfusion parameters of blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability surface area product (PS) were obtained automatically. All parameters were statistically analyzed among groups. Results TDC showed single peak in common carotid artery, with small peak of speed up and slow down in benign group, while with baseline segment, up above, down segment and horizontal segment in thyroid carcinoma. There was statistical difference between benign and malignant groups in BF, BV, MTT and PS value (P=0.001, <0.001, 0.003 and <0.001, respectively). No significant difference of BV and MTT was found between subunits of benign and thyroid carcinoma (all P>0.05). BF was significantly different in benign and maligant groups (P<0.05), whereas PS in thyroid carcinoma and nodular goiter was significant different (P<0.05). No statistical difference of BF, BV, MTT and PS was detected between nodular goiter and thyroid adenoma. Conclusion MSCTPI can exactly show the blood flow features of thyroid. The analysis of BF, BV, MTT and PS is helpful for differential diagnosis between benign thyroid disease and thyroid carcinoma.

Objective: To investigate the clinical effect of acupuncture-moxibustion therapy on simple obesity due to spleen deficiency. Method: Sixty-eight cases of simple obesity due to spleen deficiency syndrome, including internal dampness due to spleen deficiency, qi deficiency of lung-spleen, yang deficiency of spleen-kidney, were randomly allocated into two groups, treatment group (36 cases) and control group (32 cases). The former group was treated with warm needling moxibustion method, and the latter was treated with electroacupuncture. The therapeutic effect and body weight (BW) were observed, respectively. Result: There was a significant difference between the two groups in the total effective rate (P<0.05); there was a significant difference between the two groups in the decrease of BW (P<0.01), furthermore, the long-term effect in warm needling moxibusiton group was better than that in the electroacupuncture group. Conclusion: The effect of needle wanning moxibustion therapy on simple obesity due to spleen deficiency is accurate, and it is worth to spread.

Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.