Objective To study the incidence of complications of visceral and vascular injury in laparoscopic surgery for gynecologic diseases,and to discuss the ways to decrease the incidence. Methods The data of 2684 patients who received laparoscopic surgery from Januray 2003 to December 2005 in the Department of Obstetrics and Gynecology were reviewed retrospectively.The incidence and treatment of complications of visceral and vascular injury were observed. Results The total incidence rate of complications was 2.53%(n=68),and that of the visceral and vascular injury was 0.37%(n=10).Four cases of injury were related with trocar punctures(injury of omental blood vessel,n=2;postperitoneal vessel,n=2),three took place during the operation (severe bleeding,n=1;bladder injury,n=2),and the other three were observed in the postoperative stage(ureter injury,n=2;intestinal fistula,n=1). Conclusion The complications in laparoscopic surgery for gynecologic diseases are increased with the extension and difficulty of operation,and are closely related with the experience of the surgeons.Proper candidates for the surgery and established operative technique are the key factors in decreasing the incidence.

Purpose　To study the anti-inflammatory,anti-allergy and immunosuppressive effect of polyglycosides of tripterygium wilfordii Hook (TⅡ).　Methods　Animal model of acute and subacute inflammation,adjuvant arthritis,type Ⅳ and Ⅲ allergic reaction were used.Serum haemolysin level,lymphocyte proliferation and T-cell subsets in spleen were tested.　Results　TⅡ 60,120 mg/kg and pre(prednisone)10 mg/kg had marked inhibitory effect on carrageenine induced swelling,tampon induced granulation and on the prevention of adjuvant arthritis.The delayed type hypersensitivity reaction(type Ⅳ) and arthus reaction (type Ⅲ) was distinctly suppressed.It had a suppressive action on serum haemolysin and lymphocyte proliferation with and without concanavatin A which was dose dependent.In the T-cell subsets:TⅡ 60 mg/kg decreased TS and TⅡ 120 mg/kg inhibited both TS and TH,preacted mainly on TH and rendered the decreased ratio of TH/TS.　Conclusions　TⅡ has suppressive effect on inflammation,allergic reaction and immune function.

Ikaros is a regulatory factor playing an important role in control of the development of hema-topoietic cells and immune system;and as a tumor suppressor,its aberration can cause both T and B lineage development arrest and neoplastic transformation,leading to insensitive to therapy and poor prognosis in actue lymphoblastic leukemia. Recently,researches on how Ikaros affects the leukemogenesis and prognosis become the focus of attention.

Objective To design a type of poly (D, L) Lactide (PDLLA) cage, compare the characteristics of inter body fusion using PDLLA cage with those of titanium cage and autologous tricortical iliac crest graft in a goat cervical spine model in vi?vo. Methods Twenty?four goats underwent C3-4 discectomy and fusion were assigned to 3 groups, PDLLA cage group (n=8), titani?um alloy cage group (n=8) and autologous iliac bone group (n=8). Radiography was performed pre?and post?operatively and 1, 2, 4, 8, and 12 weeks after operation. At the same time points, disc space height (DSH), intervertebral angle (IVA), and lordosis angle (LA) were measured. After 12 weeks, the goats were killed and fusion sites were harvested. Biomechanical testing was performed in flexion, extension, axial rotation, and lateral bending to determine the stiffness and range of motion. All cervical fusion speci?mens underwent histomorphological observation. Results The IVA of PDLLA cage 4 weeks after operation and DSH 8 and 12 weeks after operation was statistically greater than that of autologous iliac bone graft (P<0.05). The LA values were shown no signif?icant difference among PDLLA cage, Titanium cage and autologous iliac bone graft groups. The stiffness of two types of cages in ax?ial rotation and lateral bending, the ROM in every movement was statistically greater than that of autologous iliac bone graft group (P<0.05). PDLLA cage and Titanium cage had no significant difference (P>0.05). Radiographic and histomorphological observa?tion showed better fusion results in cage groups than in autologous bone group. Conclusion This type of PDLLA cage has excel?lent biocompatibility and can provide an appropriate biological environment for bone ingrowth and osteogenesis at bone?implant in?terface. Furthermore, PDLLA cage can maintain DSH and increase the stability of fusion segments to create a good biomechanical environment for the last bone fusion.

BACKGROUND:Typical anterior cervical decompression and fusion method has many disadvantages.Cervical fusion cage has obtained satisfactory outcomes,but many problems should be solved.OBJECTIVE:To design a type of PDLLA Cage and to investigate the immediate stability of different fusion methods in a goat cervical spine model following C_(3-4) fusion.DESIGN,TIME AND SETTING:Randomized controlled study was performed at the Laboratory of Biomechanics,Sichuan University from June 2005 to January 2006.MATERIALS:C_(3-4) segment was obtained from a total of 15 adult female 2-year goat corpses.Additional C_(3-4) from 27 adult female 2-year goats was used in biomechanics test.Samples were supplied by the Animal Experimental Center,West China Clinical Medical College,Sichuan University to use in measurement of anatomy parameter and biomechanica test.METHODS:C_(3-4) segment obtained from goat corpse was subjected to X-ray test in an anterior lateral position.In accordance with C_(3-4) mean vertebral height,the angle of C_3 vertebral inferior border and anterior border,the anterior and posterior distance of C_3 vertebral inferior border,as well as the distance of anterior and posterior borders of the middle vertebral body,a type of PDLLA Cage was designed and made,and the biomechanical behavior analysis about the materials was performed.The 27 goat samples were not destroyed following biomechanics test (normal control group),and were assigned to 3 groups,PDLLA Cage group (n=9),titanium alloy Cage group (n=9) and autologous iliac bone group (n=9).Anterior longitudinal ligament was incised at FSU intervertebral space.Following excising the nucleus pulposus,adherent muscle was removed and the whole ligament was remained,as well as the bony terminal plate and cartilage subchondral bone were reserved.PDLLA Cage and titanium alloy Cage were respectively implanted in the cavity of the Cage main body in the PDLLA Cage group and titanium alloy Cage group.The Cage was fixed in the vertebral body using two fixing plates of anterior alar plate with 3.5-mm screw.Autologous cortical iliac bone from three surfaces was implanted in the autologous iliac bone group.MAIN OUTCOME MEASURES:Biomechanics was tested in flexion,extension,axial rotation,and lateral bending using 1,2,3,4 N·m moment of force method.The mean stiffness values and the range of motion were calculated in each group. RESULTS:In every loading mode,the displacement values were statistically significant difference between cage groups (PDLLA Cage group and titanium Cage group) compared with the normal control group (P<0.05);and the displacement of cage groups were lower than that of the autologous tricortical iliac crest bone graft group except for 4 N·m bending loading mode (P<0.05).Posterior extension was significant better in the PDLLA Cage group and titanium alloy Cage group compared with the autologous iliac bone group.Compared with the normal control group,the displacement of posterior extension and lateral bending was lower in the autologous iliac bone group (P<0.05).The stiffness of PDLLA Cage and the titanium Cage group in the flexion loading had no significant difference (P>0.05).In the others loading condition,the titanium Cage group was statistically greatest in all directions (P<0.05).The stiffness of PDLLA Cage was comparable with that of the titanium Cage and was statistically higher than that of the autologous tricortical iliac crest bone graft and intact (P<0.05).CONCLUSION:This type of PDLLA Cage can provide enough primary stability for cervical intervertebral fusion.

Two pairs of primer targeting to the genes encoding the major outer membrane protein (MOMP) and the polymorphic membrane protein (PMP) of Chlamydophila psittaci (Cps) designed and used in the fluorescent quantitative PCR assay in the detection of this microorganism were compared in the their sensitivity and specificity.It was demonstrated that both pairs of primer could be used successfully to detect the presence of Cps of the epidemic strains isolated in China.The specificity of PMP primer used was higher than that of MOMP primer. In case of high target concentration in samples, the sensitivity of the former was also superior to that of the latter, but the amplification efficiency of the former was lower than that of the latter. The MOMP primer seemed to be more suitable to detect Cps by using the real-time quantitative assay.As demonstrated by the real-time quantitative PCR assay, great difference in the genomic copy numbers of the PMP gene existed in the epidemic strains of Cps isolated in China.

Objective To investigate the effects of deletion of adenosine A2A receptors on peripheral leukocytes on cerebral white matter lesions induced by chronic cerebral hypoperfusion.Methods Twenty-four wild type(WT) male mice were given a ? irradiation of 12.5Gy,followed by receiving bone marrow cells tail vein from female A2A receptor knocked out(KO) mice via tail vein,were assigned as KO→WT group,while those received bone marrow cells from WT female mice were assigned as WT→WT group(n=20).The efficiency of reconstitution of bone marrow cells in recipient mice was assessed 7 weeks after transplantation by PCR and immunofluorescent technique.Then,the recipient mice were subjected to bilateral common carotid artery stenosis with internal diameter of 0.18mm by external banding using microcoils at 8 weeks after transplantation.On 7d,14d and 30d after the surgery,corpus callosum,fiber bundles of Caudoputamen and optic tract were harvested from the cerebral white matter,and stained with Kluver-Barrera staining for observing the changes in nerve fibers,and with GFAP and CD11b immunohistochemistry staining for observing the proliferation of microglia and astrocytes.Results At 7 weeks after successful transplantation,the genotype of sex chromosome in peripheral leukocytes of the male recipient mice was changed into female pattern.The expression rate of A2A receptor was 9.73%?2.05% in KO→WT group and 93.82%?11.24% in WT→WT group,with significant difference between the two groups(P

Objective To observe behavioral characteristics and pathologic changes of chronic cerebral hypoperfusion in mice. MethodsTotally 62 adult C57BL/6 mice were subjected to either sham-operation (n=31) or bilateral common carotid artery stenosis using external microcoils with an inner diameter of 0.18 mm(n=31). At 30 d after the stenosis, the animals of the 2 groups (8 mice for each group) underwent behavioral test of 8-arm radial maze. In 7, 14 and 30 d, the rest mice were sacrificed for their brain tissue samples which were examined with Kluver-Barrera staining and immunohistochemical assay for markers of microglia and astroglia, glial fibrillary acidic protein (GFAP) and CD11b. In 14 d after the model establishment, Evans blue dye extravasation test was performed for the blood-brain barrier function. ResultsThe model group made significantly more errors than sham-operated group in 8-arm radial maze test at 30 d after the surgery. White matter lesions occurred and the proliferation of activated microglia and astroglia were observed in white matter in model mice after 14 and 30 d bilateral common carotid artery stenosis. The disruption of blood-brain barrier function of model mice was indicated in the evans blue extravasation test at 14 d after bilateral common carotid artery stenosis. ConclusionCognitive impairment, white matter lesions and glial activation are successfully induced after bilateral common carotid artery stenosis in mice model of chronic cerebral hypoperfusion.

Objective To investigate the effect of adenosine A2A receptor deficiency on the ischemic neuronal injury and its potential mechanism.Methods Transient(2 h)cerebral ischemia was induced by the middle cerebral artery occlusion(MCAO)in mice.Adenosine A2A receptor knockout(A2ARKO)mice and their wild-type littermates(A2ARWT)were divided into 2-hour cerebral ischemia group,cerebral ischemia with 22-hour reperfusion group and cerebral ischemia with 46-hour reperfusion group.Cerebral infarction volume was measured by image analysis of brain sections stained with cresyl violet(CV).Brain water content was evaluated with the dry-wet weighing method.The expression of calbindin D-28k(CB)and aquaporin-4(AQP4)in ischemic brain was determined with immunohistochemical methods.Results The cerebral infarction volumes in 2-hour cerebral ischemia group,cerebral ischemia with 22-hour reperfusion group and cerebral ischemia with 46-hour reperfusion group of A2ARKO mice were lesser than those in the corresponding groups of A2ARWT mice.Compared with A2ARWT mice,A2ARKO mice had more CB,lesser AQP4 expressions and lesser brain water contents.Conclusion Adenosine A2A receptor deficiency exerts the protection against ischemic brain injury both in the acute phase and reperfusion phase,and attenuates brain edema caused by cerebral ischemia,which may be due to the inhibition of intracellular calcium overload and AQP4 expression.

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.