AFP from fetal serum and serum of patients with primary hepatic cancer were purified by affinity chromatography. Then, the lentil lectin-reactive and nonreactive variants of these purified glycoproteins were prepared by affinity chromatography with immobilized lectin. Glycopeptides and oligosaccharides were prepared from variants by protease treatment and hydrazinolysis, respectively, and subjected to carbohydrate and amino acid analysis. A small difference in the carbohydrate compositions of each variant was observed. Analysis of chemical compositions of AFP variants is useful for the differential diagnosis between benign liver diseases and primary hepatic cancer.

Objective To explore conduct disorder tendency and it's influence factors on secondary school students. Methods 669 students of grade 1 in Dezhou Vocational and Technical College were investigated by Conduct Disorder Tendency Questionnaire, Revised Version of MBI-SS and SRHMS Version1.0. Results The secondary vocational school students' scores on violation tendency, addiction tendency and aggression tendency were 11.51±4.08,5.90±2.68 and 6.68±2.63,and were significant different from vocational college students or high school students(addiction tendency:F(2, 1578)=7.48,P =0.001;aggression tendency:F(2, 1579)=24.40,P <0.001). There existed great capable difference of conduct disorder according to genders ( t(643)>3.85,P <0.001).Conduct disorder tendency of secondary vocational students' was positive correlated with their learning burnout (Pearson correlation coefficient was between 0.16 and 0.52 for each pair), but negative correlated with mental health(Pearson correlation coefficient was between -0.13 and -0.27 for each pair). The linear regression showed that some dimensions of learning burnout and self-rated mental health predicted students' conduct disorder tendency. Conclusion Conduct disorder tendency of secondary vocational students is affected by their learning burnout and mental health.

Objective To investigate the effects of valsartan on insulin resistance of type 2 diabetes in patients with Ⅰ hypertension.Methods Newly diagnosed 152 patients of type 2 diabetes with Ⅰ hypertension were randomly divided into treatment group and control group.The treatment group and control group undergone lifestyle changes( diet and adequate exercise) for two weeks.After that,the control group were given Diltiazem,while the treatment group were given valsartan,an angiotensin receptor blocker(ARB) drugs.Both groups were treated for 8 weeks.Fasting plasma glucose (FPG) and fasting plasma insulin (FINS) before and after treatment in both groups were determined.Homeostasis model assessment evaluation of insulin resistance index(HOMA-IR) and insulin sensitivity index (IAI) were carried out and compared between the two groups.Results After 8 weeks of treatment,HOMA-IR in the treatment group was significantly decreased compared with the control group ( P ＜ 0.01 ),while IAI was significantly increased( P ＜ 0.01 ).Conclusion Application of valsartan in type 2 diabetes mellitus with Ⅰ hypertension can significantly improve the level of insulin resistance.

Objective To explore the clinical significance of expression of MKK4 in osteosarcoma,and to discuss the relationship between MKK4 and osteosarcoma occurrence,development and prognosis.Methods The expression of MKK4 protein was determined with immunohistochemical SP method in 30 cases of osteosarcoma.Results The MKK4 was obviously expressed in both nucleus and cytoplasm in osteosarcoma.The expression of MKK4 in the cytoplasm of osteosarcoma was positively correlated with clinical stage (r =0.72,P =0.000),the expression of MKK4 in the nucleus of osteosarcoma had no correlation with clinical stage (r =0.44,P =0.080).MKK4 expression in the cytoplasm was higher in transfer osteosarcoma than no transfer osteosarcoma(x2 =9.69,P ＜0.01),while no obvious difference in the nucleus of osteosarcoma(x2 =1.00,P =1.000).Conclusion The MKK4 is highly expressed in the nucleus and cytoplasm of osteosarcoma,suggests that MKK4 may be related with the carcinogenesis development of osteosarcoma.The expression of MKK4 in the cytoplasm of osteosarcoma is related to the malignant,metastasis and prognosis of osteosarcoma,may be used as a potential prognostic indicator of judgment osteosarcoma.

CXCR4, a receptor for stromal cell-derived factor-1 (SDF-1, also know as CXCL12) and a member of the chemokine receptor family, has been shown to play an important role in the proliferation, migration, invasion and metastasis of breast cancer cells. CXCR4 expression is an early event in breast cancer development and may serve as a biomarker for early detection and diagnosis of breast cancer. Expression of CXCR4 has been demonstrated to correlate with metastasis and poor prognosis in breast cancer. Inhibition of CXCR4 expression may be a potential target of breast cancer treatment.

ObjectiveTo study the toxic effect of bupivacaine encapsulated in liposomes on spinal cord in rats.MethodsOne hundred and eight SD rats (200-225 g) in which intrathecal (IT) catheter was successfully implanted without complications were randomly divided into 6 groups ( n =18 each):control group (group C) ; liposome group (group L) ; 0.5％ and 1.0％ bupivacaine groups (groups B1 and B2 ) and 0.5％ and 1.0％ bupivacaine encapsulated in liposomes groups (groups LB1 and LB2 ).In groups L,B1,B2,LB1 and LB2,liposome,0.5 ％ bupivacaine,1.0 ％ bupivacaine,0.5 ％ liposomal bypivacaine and 1.0 ％ liposomal bupivacaine 20 μl were injected IT respectively once a day for 7 consecutive days,while in group C nothing was injected IT.Pain threshold was measured by mechanical stimulation of the plantar surface of hindpaw.Motor function of the hindlimbs was also assessed.The animals were sacrificed at 8 day after IT injection.The lumbar segment of the spinal cord was removed for microscopic examination,detection of neuronal apoptosis (by flow cytometry) and Fos protein expression (by immuno-histochemistry).Results1.0％ bupivacaine IT significantly increased the percentage of apoptotic neurons in group B2 as compared with control group.0.5％ and 1.0％ bupivacaine IT significantly increased the number Fos protein positive cells in group B1 and B2 as compared with group C.1.0％ bupivacaine IT induced severe histologic damage including shrinkage of nucleus and vacuole formation in mitochondria.Encapsulation of bupivacaine in liposomes significantly attenuated bupivacaine-induced increase in apoptosis and Fos protein expression and histologic damage in group LB2 as compared with group B2.ConclusionThe encapsulation in liposomes can decrease the neurotoxicity of 1.0 ％ bupivacaine administered IT in rats.

Objective To investigate the effect of liver function with portal vein occlusion (PVO) in various phases and the following restoration portal vein flow on intestinal mucosal cells. Methods Twenty-four healthy adults white Japanese rabbits were randomized into 1 control group and 2 experimental groups (according to portal vein clamping for 30 min and 45 min). Each experimental group's blood samples were collected from caval vein 1 h before operation, by the end of portal vein oc-cluded, 30 min, 60 rain after relief of portal vein blocking, then with restoration of portal vein flow for 2 h and rabbit guts were continuously cut to sections for HE, TUNEL staining and Bcl-2, Bax protein immunohistochemical staining to observe the injury of intestinal mucosa cell apoptosis and the relation-ship between the expressions of Bcl-2 and Bax. The levels of blood glutamate-pyruvate transaminase (ALT), glutamic oxalacetic transaminase (AST) were measured and compared. Results The levels of ALT, AST in the control group did not significantly change. Compared with control group, group B did not change significantly with PVO 30 rnin in liver enzyme and they were significantly increased after portal vein occlusion relief. The levels of ALT and AST were increased obviously at 45 min with PVO, then raised again. Down-regulation of Bcl-2 expression, up-regulation of Bax expression and the increased index of apoptotic cell were found in each experimental group. Conclusion It may be more safe with PVO for 30 min. But the following restoration portal vein flow will bring about ischemia-reperfusion injury that is mainly apoptosis in the small intestine. The index of apoptosis will be raised with time prolongation of PVO.

Objective To investigate the relationship between monocyte chemoattractant protein-1 (MCP-1) gene -2518G/A polymorphism and cerebral infarction(CI) of Han population in Chinese Hunan district.Methods The -2518G/A polymorphism in MCP-1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) and DNA sequencing in 162 patients with CI (CI group) and 150 normal controls (NC group). The serum level of MCP-1 in the two groups was determined by enzyme-linked immunosorbent assay (ELISA).Results In CI group, genotypic frequency of GG was 39.5%, GA was 37.0%, and AA was 23.5%. The allele frequency of G was 58.0% and A was 42.0%. In NC group, genotypic frequency of GG was 27.3%, GA was 39.3%, and AA was 33.4%. The allele frequency of G was 47.0% and A was 53.0%. The frequencies of MCP-1 -2518GG genetype and G allele in CI group were significantly higher than those in CI group (all P

Objective To prepare the anti-hUART1 polyclonal antibody and investigate the subcellular localization of hURAT1 protein. Methods The full-length hURAT1 gene was obtained by RT-PCR and inserted into the fusion expression vector pEGFP-N3,the predicated antigen epitope was cloned into GST fusion protein expression plasmid pGEX-5X-1, transformed into E. coli BL21 cells for expressing recombinant GST-hURAT1 protein induced by IPTG. The purified hURAT1-GST fusion protein was employed to immunize rabbit for preparing the polyclonal antibody. The expression of hURAT1 was analyzed by western-blot and immunohistochemistry in human kidney. pEGFP-hURAT1 was transfected into the LLC-PK1 cell in order to observe the subcellular localization of the gene using confocal microscopy. Results Specific anti-hURAT1 rabbit polyclonal antibody was obtained, and both Western-blot and immunohistochemistry showed that hURAT1 was expressed in the human kidney brush border,localized in the apical membrane of the LLC-PK1 cell. Conclusions hURAT1 protein was a membrane protein located in renal proximal tubule, which could be detected in the apical membrane. The anti-hURAT1 polyclonal antibody could be used for studying the physiological function of hURAT1 and its pathology.

Aim To investigate the effect of selective cyclooxygenase-2 inhibitor, NS-398, on cell apoptosis in leukemia cell line K562 cells and its underlying mechanism. Methods Apoptotic cell percentage was examined by flow cytometry(FCM). The protein expression of Bcl-2 and Caspase-3 was detected by Western blot, and the activity of Caspase-3 was checked by FCM. Results NS-398 induced the cell apoptosis in K562 cells in a dose-dependent manner. After NS-398 treatment, the expression of Bcl-2 protein was downregulated, whereas the expression of Caspase-3 protein was upregulated. Moreover,the activity of Caspase-3 was increased in a dose-dependent way after NS-398 treatment. Conclusion Selective cyclooxygenase-2 inhibitor,NS-398,significantly induced apoptosis in K562 cells. The underlying mechanism might be related to the downregulation of Bcl-2 and the activation of Caspase-3.