Objective To construct yeast eukaryotic expression vector carrying human endostatin (ES) cDNA. Methods The functional fragment of endostatin gene in human hepatic tissue was amplified by using RT-PCR technology, and cloned into yeast pPIC9 expression vector. The positive clone was sequenced by using automatized sequencer. Results The endostatin cDNA was successfully cloned. The positive ES clone gene in pPIC9 expression vector was sieved, and its coding sequence was identified to be as same as the previously reported sequence. Conclusion The successful construction of ES gene in pPIC9 expression vector using molecular biological method maybe helpful for the high expression of ES protein, which may lay the foundation for the treatment of malignant tumor through anti-angiogenesis appoach.

Objective:To study the effect of simvastatin therapy on bone density and metabolism in rats with osteoporosis. Methods:Forty female Wistar rats were randomly divided into four group: control group-A (sham operation group), ovariectomized group-B(OVX), OVX with oestrogen treatment group-C and OVX with simvastatin treatment group-D. Thirty days after surgery, oestrogen replacement therapy(nilestriol 1 mg/(kg?d), ig once a week for 12 weeks) in group-C and simvastatin therapy(with simvastatin 5 mg/(kg?d), ig once a day for a week, paused for two weeks,and given for another one week,until 12 weeks)in group-D.After 12 weeks, all rats were sacrificed. On the 12th day and the 4th day before sacrifice,all rats received subcutaneous injections of tetracycline as in vivo label. The proximal tibiae of rats were processed to undecalcified sections and stained with HE or Von Kossa.The microarchitecture was studied by bone histomorphometry. Results:Osteoporotic changes in proximal tibiae were observed in all ovariectomized rats and not in sham-operated rats. There was significant difference in static parameters (%Tb.Ar, Tb.N,Tb.Sp) comparing group C or D with group B respectively(P

Objective:to do neuropsychological tests and apply functional questionnaire in screening of Alzheimer disease in rural area of north China. Method: subjects positive in screening with MMSE were applied a battery of neuropsychological tests including Fuld Object Memory test (FOM), a categorical test (Animal Naming Test), Digit Span Subtest from WAIS-R, and Block Design Subtest from WISC-III. Examination of apraxia and Pfeffer Functional Questionnaire were also applied.Result:There were significant differences of FOM in different groups, such as Alzheimer disease (AD) group, vascular dementia (VD) group, depression group, and VD+AD group. FOM had better sensitivity and specificity, while the sensitivities of RVR and apraxia were low, and the specificity of Digital Span and Block Design were not high enough.Conclusion:FOM may play an important role in diagnosis of Alzheimer disease.

AIM:To observe the therapeutic effect of glucagon-like peptide 1 (GLP-1) analog on nonalcoholic fatty liver disease of rats and to investigate the underlying mechanism.METHODS:SD rats (n=21) were used to estab-lish a nonalcoholic fatty liver disease model by feeding a high fat diet for 12 weeks, and other 11 rats were fed with a normal diet for 16 weeks.The model rats were randomly divided into 2 equal groups:one group was treated with glucagon-like pep-tide 1 analog (0.6 mg· kg-1 · d-1 ) by intraperitoneal injection for 4 weeks, the other group using saline as a control.Af-ter treatment, fasting blood glucose, serum insulin, blood lipids, liver function and the pathological changes of the hepatic tissues were evaluated and the expression of PKCεat mRNA and protein levels in the liver tissues was detected by real-time PCR and Western blot, respectively.RESULTS:Compared with model group, the intervention of GLP-1 significantly re-duced insulin resistance index (HOMA-IR), improved the liver function (P<0.05), decreased the liver index and blood lipids (P<0.05).HE staining showed obvious pathological changes of the hepatic tissues in model group, and the inter-vention of GLP-1 significantly reduced lipid droplets in the hepatocytes and improved the structural damage of the liver.The expression of hepatic protein kinase Cε( PKCε) at mRNA and protein levels significantly decreased which were reversed by treating with GLP-1.CONCLUSION:GLP-1 shows good therapeutic effect on nonalcoholic fatty liver disease of rats, pos-sibly by controlling lipid metabolism and reducing insulin resistance, which may be related to PKCεexpression.

Objective: Affective disorder patients can be effectively treated by electro-acupuncture methods without many side effects, but the mechanism of the treatment is not known. The object of this study was to discover whether there is change of guanine-nucleotide-binding protein (G-protein) level in platelets membrane from major depression patients before and after treatment by fluoxetine and electro-acupuncture. Methods:We utilized selective antibodies to quantitate the levels of the G-protein ? subunits in platelets membrane of 78 major depression patients (24 received fluoxetine treatment, 24 received electo-acupuncture treatment and 30 received placebo treatment) and 30 matched comparison subjects.Result:Levels of both G?i and G?q of the patient groups were higher than matched normal comparison subjects before treatment of fluoxetine and electro-acupuncture,the level of G?i and G?q did not change after the treatment of fluoxetine, electro-acupuncture and placebo. The level of G?s wasn't significantly different with matched comparison subjects.Conclusion:The levels of G?i、G?q in platelets membrane from major depression patients are higher than normal, the high levels of G?i and G?q in platelets membrane may be a trait abnormality for major depression patients, not a state abnormality for major depression.

Objective: To investigate pathophysiological mechanism of tardive dyskinesia (TD) by comparing the level of serum prolactin ( PRL ) of schizophrenic patients with TD to that of patients without TD (TD-) and normal volunteers. Methods: To assay the level of serum PRL in normal controls (n=44), chronic schizophrenic patients with TD (n=46), and without TD (n=46) by radio-immunity assay (RIA). The TD group and TD^group were matched by age, medication,disease duration matched. AIMS (abnormal involuntary movement scale) was used to measure the severity of TD. Results: The concentration of PRL in serum in the normal group (617?358?IU/ml), patients with TD (1277?1011?IU/ml) and TD- (913?602?IU/ml) had significant difference (P0.05). Conclusion: The concentration of PRL in serum in patients with TD was increased significantly.

Objective To investigate the pathogenesis of portal hypertensive gastropathy(PHG). Methods Two rat models with portal hypertension(PHT) and a sham operation group were established to detect the pathological changes in histology and ultrastructure of the gastric mucosa as well as quantitative changes in histological morphology by graphic analysis computer system. Results Prominent edema, scattered red dots/ecchymosis were found in gastric mucosa in rats with PHT. Light and scan electron microscopy showed swelling or exfoliating of the epithelium cell, reduction of gastric gland number, thin of gastric mucosa, while infiltration of inflammatory cells, epithelium metaplasia were not found. The most characteristic findings were the changes of the mucosal capillaries and venules in both mucosal basal lamina and submucosa layer light microscopically, as well as the transmutation, stenosis and loose intercellular joining of the capillaries on electron microscopy. Ultrastructure observation revealed numerous pinocytes in epithelial cells as well as proliferation and hyperplasia of smooth muscle, collagenous fiber and extracellular matrix in venules. Quantitative analysis showed that the ratios of the cross sectional area and the vascular wall area between the gastric wall area(CSA/GWA & VWA/GWA) was higher in liver cirrhosis(LC) and portal vein stenosis(PVS) groups than that in sham operation(SO) group. There was a positive correlation between portal vein pressure and the ratio of the length of damaged mucosa and the length of mucosa(LDM/LM that was higher in LC group than in PVS group). Conclusions In rats with cirrhotic portal hypertension, distinct gastric microvascular morphological changes are the major etiological factor of PHG as a part of pathological changes in portal hypertension.

Objective:To detect the expressions of COX-2 protein and BCL-2 protein in transitional mucosa adjacent to rectal carcinoma,and determine whether the transitional mucosa was the cancer precursor event.Methods:Mucin histochemical method was employed to detect the distal mucosa 2 cm away from rectal tumor and the transitional mucosa was found in 54 cases of rectal carcinoma.Immunohistochemical method was used to investigate the expressions of BCL-2 and COX-2 protein in the specimen of rectal carcinoma mass,transitional mucosa and non-transitional mucosa,and 20 cases of normal rectal mucosa,and the points of the expressions of COX-2 protein and BCL-2 protein were counted.Results:35.19%(19/54)of distal mucosa were characterized as the transitional mucosa.The expressions of COX-2 and BCL-2 protein were detected in carcinoma mass and distal mucosa.Significant difference was observed in carcinoma mass and transitional mucosa(p<0.01).But there were no significant differences between the expressions of those in transitional mucosa and non-transitional mucosa(p>0.05),as well as transitional mucosa and normal mucosa(p>0.05).Conclusion:The performance of transitional mucosa was not the cancer precursor event,but the non-specificity reaction of carcinoma or inflammation.

Objective To investigate the expression and significance of myeloid cell leukemia-1 (Mcl-1 )gene and protein in Hepatocellular Carcinoma(HCC).Methods The expression of Mcl-1 was detected respectively by Reverse Transcription-Polymerase Chain Reaction (RT-PCR),Western blot and ENVISION immunohistochemistry in 20 HCC specimens,19 liver cirrhosis(LC)specimens,and 12 control ones.Their relationship with clinical and pathological characteristics of HCC was investigated.Results The expression of Mcl-1 mRNA in the control group,LC group and HCC group was 0.52±0.17, 3.46±1.7,3.65±2.93,respectively.The level in HCC and LC group was statistically different compared with the control group,respectively (t=7.925,5.334,P<0.05).The relative expression of Mcl-1 protein in LC group (0.51±0.35)and HCC group (0.75±0.36)were significantly higher than that in the control group (0.21±0.19)(t=5.526,6.355,P<0.05).The positive expression rate of Mcl-1 in HCC group was 55.00% (11/20),significantly higher than that in the con-trol group 33.33% (4/12)(t=7.835,P<0.05).The positive expression of Mcl-1 was related to tumor necrosis and TNM staging (χ2=4.201,P<0.05).Conclusion Mcl-1 gene and protein are differentially expressed in HCC,LC and the control, which may be involved in the occurrence,development and malignant transformation of HCC.

BACKGROUND:Pressure sensitive instrumented shoes are fast and easily used tools to measure ground reaction forces. Currently researches about the utilities of these systems in assessment of gait in children with neurological diseases have been started to run.OBJECTIVE: To find the gait characteristics of dynamic plantar pressure in children with spastic cerebral palsy. DESIGN: A cross-sectional study.SETTINGS: Department of Rehabilitation, Songgang People's Hospital; Department of Rehabilitation, Shenzhen Children's Hospital; Department of Rehabilitation Medicine, the First Affiliated Hospital of Sun Yat-sen University. PARTICIPANTS: ① Patient group: Twenty children with spastic cerebral palsy were selected from Shenzhen Children's Hospital from May 2004 to April 2005, including 9 boys and 11 girls, aged 26-66 months old, and they all could walk for more than 10 m independently. ② Normal control group: 52 healthy children with normal walking ability were enrolled, including 28 boys.and 24 girls, aged 35-76 months old.METHODS: Ultraflex gait analysis system was used to perform continuous plantar pressure tests of both groups of children. Diagram and data of dynamic plantar pressures in gait were recorded and analyzed with a computer. MAIN OUTCOME MEASURES: ① 10 sequential gait cycles were chosen from the continuous stable steps to calculate the average data with the software; ② Differences of the characteristic parameters of plantar pressure curve between the two groups.RESULTS: ① The curves of plantar pressure to time of testees in the normal control group were of the wave shapes with two peaks and one valley. Half of the CP children could not perform the two-peak shape pressure curve in the gait analysis. One kind of abnormal plantar pressure curve style was a wave shape with only one peak. Another kind of abnormal plantar pressure curve style was a shape showing asymmetry between sides and variation in different steps even in the same side. Two-peak shapes of dynamic plantar pressure were recorded in 10 CP children. ② Characteristic parameters picked up from the curves of plantar pressure had been compared between the two groups. Significant difference have been found in ts of both sides and tz1,tz2,tz3 on the right side between the two groups. It was shown that the single-limb support time of both sides in the CP children was longer than that in healthy children [left: (0.879± 0.658), (0.472±0.051) s; right: (0.818±0.682), (0.478±0.050) s; P ＜ 0.01]. The tz1 and tz3 on the right side in the CP children were longer and while the tz2 was shorter than those in the healthy children. CONCLUSION: ① Using dynamic plantar pressure gait analysis system, plantar pressure curves can be observed directly, and information about kinetic characters can be drawn straightly. ② Part of patients with spastic CP could not perform the two-peak shape pressure curves in gait analysis. In patient can be recorded of two-peak shape dynamic plantar pressure curves, ts of both sides are shorter than those in healthy children, tz1 and tz3 on the right side is longer and tz2 is shorter than those in healthy children.