Objective To observe the therapeutic role on monoclonal antibody of alkaline phosphatase(AAP) to vasogenic cerebral edema in rats(VCE).Methods 60 Wistar rats were randomly divided into 3 groups:normal group,VCE group and AAP group.VCE animal model was made by injecting AAP.Brain water content of gray and white matter were measured by Moistrue Analyzer respectively.The permeability of the blood brain barrier (BBB) was determined by Evan's blue(EB) extravasation.Results The brain water content of gray and white matter in AAP group was reduced markedly,so the permeability of BBB was reduced(P0.05). Conclusion AAP can treat VCE well. Morbidity mechanism of VCE is related to the activity of alkaline phosphatase.

Study on proteomics can explain the protein biological function of executing life activities expressed by genome.The results of the research point to a new direction for pharmaceutical and clinical development.This review focuses on the latest developments of the qualitative and guantitative researches on biological mass spectrometry in proteomics,as well as their advantages and prospects.

Objective:To investigate the microsurgical anatomy of the far lateral suboccipital approach and to evaluate its application to surgical approach. Methods:Fourteen cadaveric heads(28 sides) were dissected through far lateral suboccipital approach to obtain the anatomical data of this approach.A retrospective analysis of microsurgical treatment in 12 cases of tumors situated at lower clivus and foramen magnum was performed. Results:Transverse process of C_1 and inferior occipital trangle were important anatomical marks in the far lateral occipital approach.Drilling the condyle and jugular tubercle was benefit to expand the surgical space.Total removal of the turmors was achieved in 9 patients,subtotal removal in 3 patients,and without any surgical morbidity. Conclusion:Study on the microsurgical anatomy of the far lateral suboccipital approach was helpful in resecting tumors situated at lower clivus and foramen magnum.

Objective To construct a phage antibody Fab library of human antinuclear antibody. Methods Total RNA was extracted from peripheral blood lymphocytes of 4 patients suffering from autoallergic disease, with antinuclear IgG antibody titers in the serum higher than 1∶10 000 as estimated by indirect immunofluorescence technique. Taking oligo-dT as the primers, human immunoglobulin light chains ?/? genes as well as heavy chains Fd genes were reversely transcribed to cDNA by PCR. The ?/? light chains were initially cloned into pComb3Hss vector to construct a human recombinant light chain library, and the heavy chain genes were subsequently inserted into the corresponding sites of ?/?-pComb3Hss plasmid to generate a recombinant ?/?-Fd-pComb3Hss plasmid. The plasmid was then transformed into E. coli XL1-Blue, which was subsequently infected by the helper phage M13KO7. A random recombinant antibody library was expressed on the surface of filamentous phage. Results Human immunoglobulin ?/? light chain and heavy chain Fd genes (660bp) were amplified successfully. The light chain library with the capacity size of 2?104, and the human recombinant Fab antibody library with the capacity size of 4?104 were also successfully obtained. Finally, the phage antibody library of the Fab fragment of human antinuclear antibody, containing 2?109 CFU/ml phage, was constructed. Conclusion A phage antibody library of Fab fragment of human antinuclear antibody is constructed successfully. This research lays a foundation for the preparation for phage library of Fab fragment of human antinuclear antibody, and also paves the way for the advanced assessment for its effect on the immuno-targeting therapy of tumors.

Background and purpose:Gallbladder carcinoma, lacks of specific clinical manifestations,and is usually diagnosed at advanced stages of the diseases and even fewer can be resected.Chemotherapy has not shown significant activity in gallbladder carcinoma,so new agents should be applied to its treatment.Arsenic trioxide,verified as a breakthrough in the management of acute promyelocytic leukemia,has been applied in the research of a variety of solid tumors.This study is to investigate the biological effect of As_(2)O_(3) on human gallbladder carcinoma cell GBC cells and its mechanism.Methods:Hoechest33258 staining,DNA gel electrophoresis and flowcytometric were used to determine apoptosis.Western blot was performed to analysis apoptotic proteins expression.Results:The treatment of As_(2)O_(3) in gallbladder carcinoma cells could induce apoptosis in a dose-dependent manner and downregulate the expression of anti-apoptotic protein Bcl-2.Moreover,Bcl-2 overexpression could protect gallbladder carcinoma cells from As_(2)O_(3)-induced apoptosis.Conclusions:As_(2)O_(3) induces gallbladder carcinoma cell apoptosis via down regulation of Bcl-2.

Objective To identify the susceptibile genes in a rat model for rheumatoid arthritis (RA),to determine whether sex affects disease onset and to define the mechanisms that impacts congenic genes on arthritis. Methods Arthritis-susceptible DA rats were compared with sex/age-matched congenic rats in which alleles were substituted with alleles from arthritis resistant PVG rats. Incomplete Freund's adjuvant (IFA) was injected from the base of the tail. Arthritis was visually scored, the messenger RNA (mRNA) levels of congenic genes and cytokine were determined by reverse transcription-polymerase chain reaction. The differences between two groups were analyzed using Mann-Whitney U test. Results In oil-induced arthritis (OIA), male congenic R16 rats deviated profoundly from DA rats by decreased arthritis severity (5.9±3.8 vs 9.3±2.3, P＜0.05 ), and markedly reduced lymph node mRNA levels for calsyntenin-3 (Clstn3) gene (0.7±0.4 vs 2.2±1.6, P＜0.01 ) and interleukin (IL)-17 (1.4±2.2 vs 2.7±2.9, P＜0.05) and IL-1β (1.5±2.1 vs 2.3±2.5,P＜0.05) levels. Conclusion Rat Clstn3 gene regulates the production of pro-inflammatory cytokines of OIA in male rats. The effect of arthritis-susceptible gene Clstn3 is gender-specific.

Objective To investigate the expression of LKB1 and p53 in human gastric tissues and their correlation with clinical pathological factors.Methods The expression of LKB1 and p53 in 115 cases of gastric carcinoma and 20 cases of normal gastric tissues were detected by immunohistochemistry method,and the relation between the expression and the clinicopathological parameter of gastric carcinoma was analyzed.Results The positive rates of LKB1 in gastric carcinoma and normal tissues were 20.9 ％ (24/115) and 95.0 ％ (19/20),respectively (P ＜ 0.01).The positive rates of p53 in gastric carcinoma and normal tissues were 45.2 ％ (52/115) and 5.00 ％ (1/20),respectively (P ＜ 0.05).The analysis revealed that the high expression of LBK1 was associated with tumor lymph node metastasis,stage,Lauren classification and worse survival (P ＜ 0.05).The expression of p53 was associated with tumor lymph node metastasis,stage,distant metastasis and worse survival (P ＜ 0.05).Conclusions LKB1 protein expression may play an important role in the development and progression of gastric cancer.LKB1 may be used to assess the malignant biological behavior and prognosis of gastric cancer.

Objective To evaluate clinical and pathologic factors associated with pelvic lymph node metastasis in patients with early-stage squamous cell carcinoma of the uterine cervir.Methods From February 2004 to January 2007,135 patients with stage Ⅰ b-Ⅱ a cervical squamous cell carcinoma in Women's Hospital,School of Medicine,Zhejiang University,were retrospectively studied.The relationship between pelvic lymph node metastasis and age,clinical stage,tumor size,grade of differentiation,depth of muscular invasion,lymphatic vascular space invasion,pretreatment level of serum squamous cell carcinoma antigen,pretreatment plasma level of fibrinogen,pretreatment leveh of hemoglobin and platelet were evaluated by univariate and multivariate analyses.Results Totally 3996 lymph nodes were dissected in 135 patients,with an average of 29.6 lymph nodes in each patient.12.6%of the patients(17/135)had metastasized pelvic lymph nodes.Univariate analysis indicated that tumor size(P=0.003),depth of muscular invasion(P=0.004),vasular space invasion(P<0.01),pretreatment levels of platelet(P=0.006)and fibrinogen(P<0.01)were significantly related to pelvic lymph node metastasis.Multivariate logistic regression analysis showed that lymphatic vascular space invasion(OR:3.674,95%CI:1.825-7.393,P<0.01)and pretreatment plasma level of fibrinogen(OR:4.568,95%CI:1.779-11.725,P=0.002)were significantly related to pelvic lymph node metastasis in patients with early-stage squamous cell carcinoma of the uterine cervix.Conclusion In early-stage cervical squamous cell carcinoma,lymphatic vascular space invasion and higher pretreatment plasma levels of fibrinogen are risk factors of pelvic lymph node metastasis.

Objective to investigate the effect of berberine on the expression of AQP4 and neuronal injury after focal cerebral ischemia in diabetic rats. Methods Male SD rats were randomly divided into four groups:control(sham surgery),middle cerebral artery occlusion and reperfusion (MCAO/R),MCAO/R treated with vehicle(DMSO),MCAO/R treated with berberine. the transient focal ischemia/reperfusion was induced by in-troducing silicone-coated monofilament nylon suture from the right external carotid artery into the origin of the middle cerebral artery,which was re-moved after 60 min. In group treated with berberine,the rats were injected with berberine before and after suffered from cerebral ischemia. Similarly, in group vehicle,the animals received DMSO vehicle at the same time. the score of neurological behavior was evaluated 24 h after reperfusion. Mean-while,the rats were sacrificed for Nissl staining. to estimate cerebral edema,the wet-dry ratio was measured. the expression of AQP4 in the border of the infarct region in different groups was observed by Western blot. Results Compared with the model group,berberine improved neurological deficits(P < 0.05). Berberine treatment inhibited the neuronal deformation shown by Nissl staining(P < 0.05). Berberine significantly decreased the wet-dry ratio and reduced the expression of AQP4(P < 0.05). Conclusion these results suggested that berberine could induce neuroprotection against ischemic injury by inhibiting the expression of AQP4 in diabetic rats.

Objective To express and purify the envelope ( E) protein of Japanese encephalitis virus (JEV) in soluble form.Methods Various prokaryotic expression vectors , host strains and induction conditions including time and temperatures were screened to obtain an optimum prokaryotic expression system for JEV E protein in soluble form .The expressed protein was purified by using nickel column chromatography and gel filtration chromatography .Results The soluble JEV E protein , accounting for 23% of the totally bacteria soluble protein was effectively expressed by using the recombinant plasmid PBCX -E406 at low tem-perature.The purity of the expressed protein reached up to 85%after the purification by using nickel column and gel filtration chromatography .Conclusion Soluble JEV E protein was successfully expressed and puri-fied in a simple and efficient way .It would provide a useful tool for further investigation on JEV infection , attenuation mechanism of JE live vaccine strain SA 14-14-2 and the quality control of JE vaccine .It can also be used for the development of diagnosis assay for JEV .