Hirudin (HV) is known as the most potent and specific inhibitor of thrombin. Although hirudin has many advantages , it has the bleeding side effect and this is the great shortage of hiudin for clinical application. In order to alleviate bleeding side effect of hirudin, fusion protein, named as FHV (fusion hirudin linked with FXa recognition peptide) was designed. The fusion protein gene ( fhv) was cloned into plasmid pPIC9K. FHV engineered Pichia pastoris containing high copies was chosen for fermentation and purification at 30 L fermentor scale, finally, FHV with purity of above 97% was obtained. To investigate the function of FHV in vivo, mouse tail thrombosis model was used. In the mice thrombus tail model induced by carrageenan, FHV decreased the length of tail thrombus significantly, similar to that of HV control, and had no obvious effects on the TT, PT and APTT. In conclusion, FHV is constructed and expressed in yeast. FHV fusion proteins is obtained by fermentation and purification. FHV has antithrombotic effects not influencing IT, PT and APTT after administration immediately in animal models. Therefore, FHV is a promising anticoagulant and antithrombotic drug.

Objective To evaluate the overall effect of transsphenoidal microsuegery for 23 patients with non-adenomas disease intrasellar region.Methods A toal of 23 patients with non-adenomas disease in- trasellar region,7 cases of Rathke's cleft cyst,3 cases of craniopharyngiomas,2 cases of meningiomas,2 ca- ses of pituitary tuberculous granuloma,5 cases of pituitary abscess,2 cases of empty sella,2 cases of chord- mas,were treated via sublabio-septo-sphenoidal microsurgery.Results There were among the 23 patients, groos total removal of the disease were achieved in 14 cases,subtotal removal in 7 cases,and partial removal in remain 2 cases.Diminished visual activity and visual field defects were improved in 13 cases.Menstrual disorder in all female cases and sexual disturbance in male were improved.There was no death in group. Conclusion Microsurgical technique via transsphenoidal approach is a safe and effective one for the treatment of patients with non-adenomas disease intrasellar region.

Dibenzothiophene (DBT) was used as a model compound. A bacteria strain, which can degrade dibenzo-thiophene efficiently, was obtained. This strain was identified as Pseudomonas stutzeris UP-1 according to its morphological, physiological and biochemical characters, and 16S rDNA sequence. The strain exhibits strong degradation capacity of DBT, and the end product of degradation is a kind of soluble compound. After the analysis of product of DBT degradation, it was deduced that the degradation of DBT by Pseudomonas stutzeri UP-1 is in accordance with the Kodama mechanism.

Objective To establish the 2 day-old SD rats model of periventricular leukomalacia(PVL).Methods Sixty-five healthy 2 day-old SD rats were randomly divided into 3 experimental PVL groups and 3 control groups.The experimental PVL groups were subjected to right carotid ligation(RCL),and then they were suffered from hypoxia by 6% oxygen and 94% nitrogen for 4 hours.Meanwhile sham surgeries were performed on control groups without exposed to hypoxia.Light and electronic microscopy were used to observe brain pathological changes.Immunohistochemistry methods were used to detect the distribution and expression of glial fibrillary acidic protein(GFAP),?-amyloid precursor protein(?-APP),myelin basie protein(MBP) and O4 of brain at 72 hours post-operation.Han-(ging) test,inclined plane test,open field test and cylinder test were performed on the rats 28 days post-operation.Results 1.In the PVL groups,light and electronic microscopy showed that tissue necrosis was observed in the periventricular white matter area at early stage,and at later stage right ventricular dilation,decrease of the corpus callosum area and loss of medullary sheath were detected.The morphometrical analysis showed that GFAP and ?-APP integrated optical density(A) of PVL group was increased,and mean diameter of GFAP-immunoreactive cells was also increased in PVL group,while MBP A of PVL group was decreased compared with contral group.The density of O4-immunoreactive abnormal cells was dramatically increased in PVL group.The outcomes of neurobehavioral tests of PVL group were greatly abnormal compared with control group.Conclusion The changes in 2-day-old SD rats,RCL-hypoxia model are accor-(ded) with the pathology and behavior characeriatis of PVL.

Transient forebrain ischemia-reperfusion(I/R);Brain-derived neurotrophic factor(BDNF);Histone deacetylase 3(HDAC3);Hippocampus Objective To evaluate the effect of transient forebrain ischemia-reperfusion(I/R)on the binding of brainderived neurotrophic factor(BDNF)promoters to histone deacetylase 3(HDAC3)in the hippocampus of rat and investigate its mechanism.Methods The I/R model of SD rats(I/R group)was established by Pulsinelli four-vessel clamping method,and sham operation group(Sham group)was set at the same time,which were observed for the survival of neurons in the hippocampus of rats by Nissl staining,detected for the binding of BDNF promoters(Bdnf-p1,Bdnf-p2,Bdnf-p4 and Bdnf-p6)to HDAC3 by chromatin immunoprecipitation(ChIP)and determined for the expression of brain derived neurotrophic factor antisense(BDNF-AS)by qPCR.Results Compared with Sham group,the quantity of neurons in hippocampal CA1 region of rats decreased significantly in I/R group,while those in CA3 region and DG region showed no significant changes.The binding levels of Bdnf-p1 and Bdnf-p2 to HDAC3 in hippocampal CA1 region decreased significantly in I/R Group(t = 2.575 and 2.241 respectively,each P<0.05),while there was no significant difference in the binding levels of Bdnf-p4 and Bdnf-p6 to HDAC3(t = 1.033 and 0.348 respectively,each P>0.05);The binding levels of Bdnf-p1 and Bdnf-p2 to HDAC3 in CA3 region increased significantly(t = 12.600 and 3.191,P<0.001 and<0.05,respectively),while the binding level of Bdnf-p6 to HDAC3 decreased significantly(t = 4.029,P<0.05)and no significant difference was observed in the binding level of Bdnf-p4 to HDAC3(t = 0.175,P>0.05);In DG region,the binding level of each BDNF promoter to HDAC3 showed no significantly difference(t = 0.630 ～ 1.687,each P>0.05).Meanwhile,the expression level of BDNF-AS in hippocampal CA1 region of rats decreased significantly(t = 2.560,P<0.05),but increased significantly in hippocampal CA3 and DG regions(t = 3.543 and 3.637 respectively,each P<0.01)in I/R group.Conclusion I/R showed a significant effect on the binding level of BDNF promoter to HDAC3 in rat hippocampus,which may play a role by changing the expression level of BDNF-AS.

OBJECTIVETo study the effect on apoptotic of CD4+ T, CD19+ B in spleen of BXSB mice with systemic lupus erythematosus treated with Langchuangjing Granule and to probe into the mechanism of treatment.

METHODSThe apoptosis was examined by the flow cytometric analysis and immunofluorescence double-staining method.

RESULTSApoptosis of male BXSB mice speeds up. Langchuangjing Granule can restrain the excessive apoptosis of CD4+ T and CD19+ B cells in spleen.

CONCLUSIONLangchuangjing Granule treated systemic lupus erythematosus by restraining the excessive apoptotic of T, B lymphocytes, probably restraining the release of excessive amount of apoptotic DNA fragments, so decreasing abnormal proliferation of B cells and the produce of autoantibodies.

Animals ; Antigens, CD19 ; analysis ; Apoptosis ; drug effects ; B-Lymphocytes ; pathology ; CD4-Positive T-Lymphocytes ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Lupus Erythematosus, Systemic ; immunology ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Random Allocation ; Spleen ; pathology

OBJECTIVETo investigate the expressions of stromal cell-derived factor-1 (SDF-1) and CX chemokine receptor-4 (CXCR-4) in patients with hepatocellular carcinoma (HC) and liver cirrhosis.

METHODSPeripheral blood and/or ascites fluid were collected from 39 hepatocellular carcinoma patients, 16 patients with liver cirrhosis, 12 with hepatitis and 12 healthy donors. The SDF-1 expression was assayed by ELISA and CXCR-4 was measured by immunohistochemical methods.

RESULTSThe level of SDF-1 expression in the carcinoma patients was higher than that of the liver cirrhosis, hepatitis patients and healthy donors, but there was no significant difference between those of the healthy donors and hepatitis patients or liver cirrhosis patients. The levels of CXCR-4 expression were closely related to the tumor differentiation.

CONCLUSIONThe expression of SDF-1 in the peripheral blood and the CXCR4 expression in the HCC tissues of the HC patients may be regarded as markers of HC and they may have a positive relationship with the differentiation and metastasis of HC.

Adult ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Case-Control Studies ; Chemokine CXCL12 ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; pathology ; Neoplasm Staging ; Receptors, CXCR4 ; metabolism

OBJECTIVETo investigate the autoimmune injuries of diabetic macrovascular disease (aorta) and the protective effects of immunosuppressive agent (cyclosporine A, CsA) on aortic injuries in streptozotocin (STZ)-induced diabetic rats.

METHODSSTZ-induced diabetic rats were assigned randomly to 6 groups which received low (BML or AML, 1 mgxkg(-1)xd(-1)), middle (BMM or AMM, 4 mgxkg(-1)xd(-1)) or high (BMH or AMH, 8 mgxkg(-1)xd(-1)) dose of CsA from 1 week before or after STZ for 8 weeks. Diabetic rats without any treatment, insulin-treated diabetic rats and normal rats were also monitored simultaneously and served as control groups. The pathologic abnormalities of the aorta were verified by HE, Masson staining and electronmicroscopy. The depositions of immunoglobulins (IgG, IgM and IgA) were determined by immunohistochemistry and immunofluorescence methods.

RESULTSAt the end of study, lymphocytes infiltration and collagen content (26 582 +/- 6901) were significantly higher in diabetic aorta than those in non-diabetic aorta (Collagen: 7482 +/- 3491, P < 0.01). The deposited IgG and IgA were also significantly increased in diabetic aorta compared with non-diabetic aorta (IgG: 11 789 +/- 2491 vs. 2518 +/- 1066, P < 0.01; IgA: 17 430 +/- 3159 vs. 1135 +/- 758, P < 0.01). These changes were not affected by insulin while CsA intervention significantly reduced aortic collagen content (BMH: 13 518 +/- 5440, P < 0.01 vs. STZ) and immunoglobulin deposition (BMH: IgG: 7584 +/- 4462; IgA: 6176 +/- 1900, all P < 0.01 vs. STZ). These immunoglobulin deposition changes were confirmed by results of immunofluorescence. Aortic collagen accumulation was positively correlated to aortic immunoglobulin deposition (IgG, r = 0.556, P < 0.01; IgA, r = 0.661, P < 0.01).

CONCLUSIONSOur data suggest that the autoimmune injuries might be a promoting factor in the pathogenesis of the diabetic macrovascular disease which could lead to the development of macrovascular disease. Immunosuppressive agent, such as CsA, could inhibit the abnormal deposition of immunoglobulins and therefore, delay the development of diabetic macrovascular disease in this model.

Animals ; Aorta ; immunology ; pathology ; Aortic Diseases ; etiology ; Cyclosporine ; pharmacology ; Diabetes Mellitus, Experimental ; immunology ; pathology ; Endothelium, Vascular ; drug effects ; pathology ; Immunosuppressive Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo bring forward a method to improve the prominence of the forehead contour.

METHODS16 patients(age 29-53 years, female 11, male 5) were treated through a bicoronal incision. Following the incision, the forehead soft tissues are reflected down to the level of the superior orbital rims with the protection to the supraorbital neurovascular bundles. To adjust the forehead contour, both superior orbital rims and frontal bossing was reduced by burring down the excess bone or removing the anterior wall of the forehead.

RESULTSThe operative effect were satisfactory during 3-12 months of follow-up. No recurrence or osteogenesis were found in the cases. Nasofrontal angle and forehead angle were normal. The prominence of the supraorbital ridges disappeared and forehead became nature.

CONCLUSIONCranioplasty via an osteomy or burring of the outer line of forehead bone are safe, compact and effective way to improve the prominence of the superior orbital rims or frontal bossing.

Adult ; Female ; Forehead ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods

Objective To evaluate the changes of nerve conduction velocity in degenerative rabbit sciatic nerve authograft induced by 60Co irradiation after orthotopic replantation. Methods A 30-mm-long segment was severed from normal adult rabbit sciatic nerve and exposed to 60Co irradiation at the dose of 350 Cry to induce neural degeneration. The nerve segment was then replanted orthotopicaily, and the nerve conduction velocity was determined using electrophysiological test at 4, 6 and 8 months after the replantation. Results At 6, 8 months after the replantation, the nerve conduction velocity in the degenerative nerve autograft showed no significant difference from that in normal sciatic nerve (P>0.05). But at 4 months after the replantation, the nerve conduction velocity in the autograft was significantly lower than the normal velocity (P<0.05). Conclusion The nerve conduction velocity can be obtained by replantation of a long (3 mm) degenerative nerve segment due to 60Co irradiation.