ObjectiveTo research collateral pathways and collateral grading through digital subtract angiography(DSA) and their relation to clinical prognosis.MethodsCollateral pathways and collateral grading of 49 cases suffered from severe internal carotid artery (ICA) stenosis (70% -99% ) were assessed through DSA.Incidence of stroke,TIA or cerebral hemorrhage were observed in the following 12 months.Results①In all the subjects,no collateral pathway exist in 9 cases(18.9%),and primary and secondary collateral pathways occurred in 39(79.6% ) and 25(51.0%)cases.Pathway is not related to clinical prognosis.②There were 9 ,15 and 25 cases in 0 grade,1 -2 grade and 3 -4 grade collateral circulation and the complication ratio is 44.4%,50.66% and 24.00% respectively.Incidence of ischemic cerebrovascular disease is significantly higher in 3 - 4 grade patinets than those in 0 and 1 - 2 grade pation(x2 =4.856,3.242;all P ＜ 0.05 ).ConclusionsCollateral levels but not pathway were correlated with clinical prognosis in patients with severe stenosis of internal carotid artery.

Objective To investigate the expression of Semaphorin 3F (SEMA3F) protein in hepatocellular carcinoma (HCC) and to demonstrate its relationship with clinicopathological features and prognosis of HCC. Methods Western Blotting was carried out in 32 hepatocellular carcinoma samples and matched perineoplastic tissues to detect the expression of SEMA3F protein. The relationship between SEMA3F protein expression and clinicopathological features as well as prognosis of HCC patients was analyzed. Immunohistochemistry was used to show the location of SEMA3F in HCC cells and its relationship with microvessel density (MVD). Results The expression of SEMA3F protein in HCC tissues was significantly higher than in the perineoplastic tissues (447.78± 48.26 vs 618.93 ±61.23, P＜0. 05) and it was correlated closely with tumor capsulation and tumor nodular number (P＜0.05). Based on the Western Blotting and clinical follow-up data, we found that the survival time of HCC patients with a higher SEMA3F expression level was longer than those with a lower level, and the recurrent/metastatic time of HCC patients was significantly different between these two groups (P＜0.01). Immunohistochemistry demonstrated that SEMA3F protein localized in the cytoplasm of HCC cells and its expression correlated with HCC MVD. MVD in the low-level group was higher than the high-level group (115.6±30.38 vs 86. 56±17.94, P＜0.01). Conclusions SEMA3F expression in HCC was significantly down-regulated and correlated closely with tumor-capsulation, nodular number, and MVD, implicating SEMA3F may play an important role in recurrence and metastasis of HCC. It can be regarded as a prognostic marker in HCC patients.

Objective To investigate the changes of phenotypes and function of CD56+T cells during primary HIV-1 infection and their relationship with disease progression.Methods Peripheral blood mononuclear cells (PBMCs) were collected from 53 subjects with primary HIV-1 infection and 31 HIV-1-negative healthy subjects.The percentages of CD56+T cells and the expression of several phenotypic markers on CD56+T cells including CD16, CD161, NKB1, NKG2A, NKp46, NKG2D, NKG2C and CD158a were analyzed by flow cytometry.IFN-γand TNF-αreleased by CD56+T cells with and without K562 stimulation and the levels of cytotoxic molecular CD107a were measured.Results The percentages of CD56+T cells in patients with primary HIV-1 infection were significantly lower than those of healthy subjects (P=0.025). The levels of CD56+T cells were negatively related to the viral loads in plasma samples ( P=0.021, r=-0.316).Compared with healthy subjects, the expression of CD16 (P=0.003), CD161 (P=0.023), NKB1 (P=0.023) and NKp46 (P=0.021) on CD56+T cells were decreased in patients with primary HIV-1 infection.The levels of NKB1 were positively related to the CD4+T cell counts ( P=0.007, r=0.364), but were negatively related to the viral loads in plasma samples (P=0.030, r=-0.299).Sponta-neous secretion of IFN-γand TNF-αby CD56+T cells and the expression of CD107a were dramatically in-hibited in patients with primary HIV-1 infection as compared with healthy subjects ( all P<0.001 ) . Moreover, the killing ability of CD56+T cells against K562 target cells was weakened in patients with prima-ry HIV-1 infection as the levels of IFN-γ-, TNF-α-and CD107a-producting CD56+T cells were significantly decreased (P<0.001 for IFN-γand TNF-α, P=0.016 for CD107a).Conclusion Inhibited expression and altered phenotypes of CD56+T cells were identified during primary HIV-1 infection.Lower levels of cy-tokines and cytotoxic molecular were also detected, indicating the dysfunction of CD56+T cells appeared dur-ing early stage of HIV-1 infection and was associated with disease progression.

Objective To investigate the impact of HLA-A antigens carrying Bw4 epitopes on disease course of HIV-1 infection.Methods Three hundred and forty subjects chronically infected with HIV-I were recruited and their HLA-A and B alleles were genotyped with sequence-based high resolution typing assay.HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD+4 T cell counts and viral load were analysed.Results When compared with subjects canting no Bw4 epitopes in HLA-A and HLA-B loci (OBw4) (median of CD+4 T cell counts:294/μ1;plasma viral load median 6.29×104 copies/ml),CD+4 T cell counts in subjects with genotypes of 1Bw4-A and 2Bw4-AA were comparable (307 and 308/μ1,respectively),but higher viral load (1.53×105 and 2.68×105 copies/ml,respectively) was observed.In subjects with Bw4 epitopes in HLA-B alleles but no in HLA-A,significantly higher CD+4 T cell counts (417/μ1,P=0.013) and lower viral load (2.10×104 copies/ml,P=0.007) were observed compared with those without Bw4 in HLA-A and HLA-B.Conclusion HLA-B antigens carrying Bw4 epitope were protective in HIV-1 infection by maintaining higher CD+4 T cell counts and lower viral load,but such protective effect was not observed in HLA-A antigens earring Bw4 epitope.

Objective To explore the impact of broad antigen HLA-Bw4 on disease progression in HIV-1 infected subjects. Methods Three hundred and forty subjects chronically infected with HIV-1 and 69 HIV-1 negative subjects were recruited and HLA-B alleles were typed with sequence-based high resolution typing assay. HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD4+ T cell counts and viral loads were analyzed. Results Sixty-five HLA-B alleles were detected in HIV-1 positive subjects. Subjects with Bw4 (Bw4 homozygotes and Bw4Bw6 heterozygotes ) had higher CD4+ T cell counts ( P = 0. 004 ) and lower plasma viral load ( P = 0.003 ) than subjects without Bw4 ( Bw6 homozygotes). When compared with HIV-1 postive subjects with CD4+ T cell counts above 500 celis/μl, those with CD4+ T cell counts below 500 cells/μl were observed with decreased percentage of Bw4Bw6 heterozygote ( P =0.0002) and increased percentage of Bw6 homozygotes ( P < 0. 0001 ). There is no significant difference in CD4+ T cell counts between Bw4 homozygotes and Bw4Bw6 heterozygote, but lower viral loads were observed in Bw4Bw6 heterozygotes( P = 0. 037 ). Conclusion HLA-Bw4 can confer pretective effects on H1V-1 infected subjects by maintaining higher CD4+ T cell counts and lower viral load, the mechanism behind this effect need further exploration.

Objective To investigate the diagnostic value of macrophage migration inhibitory factor (MIF) for hepatocellular carcinoma (HCC).MethodsThe research was divided into 2 parts,including testing research and confirmatory research.The clinical data of 269 patients with HCC ( group A) and 390 individuals (including 135 patients with hepatic cirrhosis,106 with benign hepatic diseases and 149 healthy individuals,control group A) who were admitted to the Zhongshan Hospital of Fudan University from January to May,2004,and 173 patients with hepatic cancer (group B) and 257 individuals (including 86 patients with hepatic cirrhosis,79 with benign hepatic diseases and 92 healthy individuals,control group B ) who were admitted from August to December,2004,and 80 patients with HCC who received radical hepatic resection in January 2005 were retrospectively analyzed.Samples of plasma of patients in the group A and individuals in the control group A were collected before operation.Samples of plasma of patients received radical hepatic resection were collected preoperatively and at postoperative day 3,7 and 30.HCC and adjacent issues of patients in the group A were collected.The levels of MIF in the plasma and tissues were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry,respectively.Non-normal distribution data were described as M( QR).Differences between the groups were analyzed by using the Mann-Whitney U test,and the relationship between the levels of MIF in the plasma and tissues was detected by the Spearman correlation coefficient.The diagnostic value of MIF was analyzed by the ROC curve.ResultsThe levels of MIF in the plasma of patients in the group A and individuals in the control group A were 85.7 μg/L (58.8 μg/L) and 15.5 μg/L(31.6 μg/L),respectively.The levels of MIF in the plasma of the patients with hepatic cirrhosis,benign hepatic diseases and healthy individuals were 24.9 μg/L (12.6 μg/L),12.5 μg/L(7.3 μg/L) and 13.2 μg/L (7.7 μg/L),respectively.There was a significant difference in the level of MIF between the group A and the control group A (F =54.235,P ＜ 0.05 ).The area under the ROC curve reached peak when the level of MIF in the plasma was 35.3μg/L.Compared with the control group B,the vdues of AUC,sensitivity and specificity were 92.1％,90.7％ and 93.4％ in the group B.The levels of MIF of the patients with HCC before operation and at 3,7,and 30 days after operation were 81.0 μg/L(54.0 μg/L),76.1 μg/L(47.5 μg/L),50.9 μg/L (40.7 μg/L) and 18.7 μg/L ( 15.1 μg/L),respectively.The levels of MIF decreased with time passed by,and were back to normal at 30 days after the operation.The median expressions of MIF in the HCC and adjacent issues were 0.083 and 0.007,respectively,with a significant difference ( U =3.975,P ＜ 0.05).The expression of MIF in the plasma was positively correlated with its expression in the HCC tissue ( r =0.759,P ＜ 0.05 ).ConclusionMIF plays an important role in the genesis and development of HCC and has potential to be one of the molecular markers for the diagnosis of HCC.

Objective By analyzing the expression of cluster of differentiation 74 (CD74) in hepatocellular carcinoma (HCC) and HCC cell lines,the correlation between the level of CD74 expression and the patients' prognosis was investigated.MethodsThe expression of CD74 in high metastatic potential HCC cell lines(MHCC-LM3,MHCC-97H),low metastatic potential HCC cell line( MHCC-97L),and no metastatic potential HCC cell line(Hep-G2) were estimated by Western blot.The paraffin embedded tissues which include intra-tumor and paratumor tissues were collected from 320 patients who had received HCC curative surgical resection and 5 normal liver tissues from the donors of liver tranplantation.The high density tissue micro-array was made of these specimens. Immunol-histochemistry was applied to discover the different levels of CD74 in tumor,paratumor and normal liver tissues.Survival curves were generated by the Kaplan-Meier method and verified by the Logrank test.Cox proportional hazards regression analysis was applied to estimate the prognostic factors in multivariate analysis.ResultsThe expression level of CD74 was significantly higher in low metastatic potential and no metastatic potential HCC cell lines (MHCC-97L 1.224 ±0.014,Hep-G2 1.374 ±0.006) than that in high metastatic potential ones( MHCC-LM3 0.622 ±0.078,MHCC-97H 0.732 ± 0.083 ).Significant differences can be found between the groups (t =- 13.308,- 16.849,- 10.177,- 13.436,- 17.057; P ＜0.01 ).Meanwhile,in tumor tissues,the CD74 was expressed positively in 221 patients and negatively in 99 patients.But CD74 was expressed slightly in paratumor and negatively in 5 normal liver tissues.There's no significant differences between the groups categorization according to age,HBsAg,cirrhosis,AFP level,tumor number,tumor size,tumor capsule,blood vessel invasion,Edmondson Grades and tumor nodes metastasis classification (TNM) stages (x2 =0.053,0.141,1.200,0.000,0.277,1.975,0.263,1.044,0.000,0.433 ; P ＞ 0.05 ),except gender (x2 =3.954,P ＜ 0.05).Kaplan-Meier method showed that patients with positively expression of CD74 had better prognosis than others (x2 =5.620,P ＜ 0.05 ).Cox proportional hazards regression analysis showed that CD74 was a significant and independent prognostic factor of survival [ hazard ratio (HR) =0.721,95％confidence interval (CI) =0.522 - 0.996,P ＜ 0.05 ].Conclusion The expression of CD74 in hepatocellular carcinoma could be a biomarker of the prognosis and there's some potential correlation with cancer cell apoptosis.

Objective To investigate the causative mutations of CYP27A1 gene in a sporadic cerebrotendinous xanthomatosis patient. Methods Genomic DNA was extracted from peripheral blood of the patient and her parents. All exons and splice sites of CYP27A1 gene were amplified by polymerase chain reaction (PCR) followed by Sanger sequenc-ing. 105 healthy unrelated subjects were also sequenced for the novel mutation in CYP27A1. Results A novel splice site mutation c.446+1G>T, a novel missense mutation c.877A>T(p.Met293Leu) and a known missense mutation c.1016C>T (p.Thr339Met) of CYP27A1 gene were identified in the patient. The mother carriers the two novel mutations and the fa-ther the c.1016C>T(p.Thr339Met) mutation. The two novel mutations were absent in 105 control subjects, respectively. Conclusions Our study detected two novel mutations, c.446+1G>T and c.877A>T, as well as a known mutation c.1016C>T, of CYP27A1 in a sporadic cerebrotendinous xanthomatosis patient. Our data provide novel information for the mutational spectrum of the gene, which is applicable in the genetic testing and diagnosis. The data also provide in-sight into the pathogenesis of the disease.

Objective To investigate clinical effect of surgical repair and reconstruction of traumatic floating shoulder injuries (FSI). Methods Sixteen patients with FSI were treated with open re-duction and reconstructive internal plate fixation. Of all, one patient was with transverse clavicle fracture, six with oblique clavicle fracture and nine with comminuted clavicle fracture. There was one patient with type Ⅰ scapula fracture, three with type Ⅱ, four with type Ⅲ, six with type Ⅳ and two with type Ⅴ. The combined injuries included rib fracture plus hemopneumothorax in six patients, traumatic moist lung in two, craniocerebral injuries in two, hepatic and splenic rupture in two and brachial plexus damage in one. The average time from primary injury to operation was seven days (3-21 days). Results Fifteen out of 16 patients were followed up for 6-28 months (average 11 months). All patients obtained bone u-nion, without infection, loosening or breakage of internal fixation. According to Herscvici evaluation standard of function, the shoulder joint outcome was rated as excellent in 11 patients, good in three and fair in one, with excellence rate of 93%. Conclusion The surgical repair and reconstruction of FSI can, to a greatest extent, recover the anatomical structure and stability around the shoulder joint and shorten the immobilization period of joint, which is beneficial to functional restoration of the joint and de-crease of occurrence of complications.

Objective To report the experience of using rectus abdominis musculocutaneous flap after open-window thoracosto- my in managing refractory chronic pleural empyema.Methods From Nov.2004 to Mar.2006,intrathoracic transplantation of the musculocutaneous flap was performed successfully in 4 patients with empyema and fistula after upper lobectomy.The myocutaneous flap was designed in such a way that epigastrica vessels were incorporated.Results The rectus abdominis myocutaneous flap has provided sufficient bulk for tract obliteration.Over a mean follow-up period of 10 months,patients are free from recurrent infections and skin nacrosis.Conclusion Vaseularized museulocutaneous flap may help in healing refractory empyema.