Objective　To discuss the development and the invasive transfer mechanism.Methods　Cell culture method, p53 protenin immunohistochemistry, PCR-SSCP examination, zymography, PCM and heterogenic inouclation test inside the nude mice were adopted. Results　p53 genetic mutation existed in human melanoma cells. The fluorescent positive rate of 67KD LN-R on the surface of the melanoma cell and the average flurescent intensity are repectively WM451＞WM983A＞WM1341B＞WM35.Early WM35 did not produce MMPs; WM1341B only produced 72KDa(MMP-2),not 92KDa(MMP-9);Both WM983A at progressive stage and distal transfering tumor WM45 produced 72KDa and 92KDa. WM983A and WM451 were also seen to form evident transfering tumor inside the naked mice.Conclusion　p53 genetic mutation, the production of MMPs and the high indication of 67KD LN-R on the celluar surface are the key factors for the human melanoma cell to obtain invasive transferring abililty.

ObjectiveTo evaluate the fourth-generation ultrasonic lithotripsy system for treating the urinary tract stones.Methods243 cases of urinary tract stones who received treatment of fourth-generation EMS ultrasonic lithotripsy system were analyzed retrospectively.ResultsImmediate phase I lithotrotrisy was performed successfully in 227 cases and the successful rate of phase I was 93.3％ (227/243).Delayed phase Ⅱ lithotripsy was performed for 16 patients.Complications happened in 2 cases,one case occurred hydropneumothorax during operation,required indwelling thoracic drainage tube processing,the other had severe bleeding which conservative treatment was ineffective,cured by the intervention of super-selective renal artery embolization.Conclusion Fourth generation of ultrasonic lithotripsy for treating stones on different anatomical locations of urinary system was safe,practical and efficient,worthy of clinical application.

Objective To establish a rapid method to detect mutations in rpoB genes of rifampin-resistant Mycobacterium tubereulosis in dinical specimens using Real-time fluorescence PCR molecular beacon assay.Methods 174 strains of Mvcobacterium tuberculosis clinical isolates were analyzed using real-time fluorescence PCR molecular beacon assay foilowed with DNA sequencing while 12 strains of NTM and 4 strains of bacteria other than Mycobacterium tuberculosis were used as the contrast.Results Eighty-two 89.1 of 92 rifampin (RIF)-resistant strains and 3 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using real-time fluorescence PCR-molecular beacon assay.The specificity, sensitivity,and accuracy of this assay were 96.3%,89.1%,and 92.5%,respectively-Eithty-three of 92 RIF-resistant strains and 1 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using the direct DNA sequencing.The specificity,sensitivity,and accuracy of the direct DNA sequencing were 98.8,90.2%,and 94.2%,respectively.As compared with real-time PCR molecular beacon assay,171 of 174(98.3%)strains of myeobactefium tuberculosis clinical isolates had the salne results.Conclusion Real-time fluorescence PCR-molecular beacon assay can be used as a rapid screen method to detect RIF-resistant isolates.

Purpose To investigate the application value in diagnosis of pleural effusion by cell block combined with immunohisto-chemistry. Methods 60 cases of pleural effusion were collected, and paraffin-embedded cell block was prepared and immunohisto-chemistry was used to detect the expression of CK7, TTF-1, E-cadherin, CEA and Calretinin. Results By use of cell block combined with immunohistochemistry, malignant detected rate was higher than that of the conventional centrifugal smear. There was statistical significance in the expression of CK7, TTF-1, E-cadherin, CEA and Calretinin between pleural effusion lung adenocarcinoma and reac-tive hyperplasia mesothelial cells (P<0. 05). CK7, TTF-1, E-cadherin and CEA were highly expressed in pleural effusion of lung ad-enocarcinoma cell. Calretinin was highly expressed in hyperplastic mesothelial cells. Conclusion Cell block and immunohistochemi-cal technique combination in the differential diagnosis of difficult pleural effusion has important clinical significance. It is worthy of popularization and further clinical application.

Objective To investigate the effect of interleukin-6 (IL-6) on the growth of human lung cancer in vivo as well as in vitro.Methods To examine the mRNA level of IL-6 receptor (IL-6R) in high-metastatic human lung giant cell carcinoma cell line PG by means of reverse transcription polymerase chain reaction (RT-PCR). To assess the existence of IL-6 receptor complex (including IL-6R and gp130) with the treatment of PG cells by use of recombinant human IL-6 (rhIL-6), recombinant human oncostatin M (rhOSM), and recombinant human leukemia inhibitory factor (rhLIF), respectively. To detect the expression of IL-6 by Northern blotting hybridization and bioactive assay. To identify the effect of IL-6 secreted by PG cells by use of IL-6 and IL-6R antisense oligodeoxynucleotides (ODNs), and specific neutralizing antibody to IL-6. To document the influence of IL-6 on PG cells growth in vivo through the strategy of the transfection of expression vector inserted antisense IL-6 cDNA.Results RT-PCR analysis revealed that PG cells expressed IL-6R mRNA. Any one of the recombinant cytokine IL-6, OSM and LIF stimulated the growth of PG cells in vitro in a concentration-dependent manner. These results demonstrated IL-6 receptor complex exist in PG cells. At the same time, PG cells expressed IL-6 mRNA and secreted bioactive IL-6. Both IL-6 antisense ODNs and IL-6R ODNs inhibited PG cells proliferation. Treatment of PG cells with IL-6 antibodies reduced the growth of PG cells in vitro. PG cells transfected with IL-6 antisense expression vector showed a decreased growth in nude mice.Conclusion IL-6 functions as an autocrine growth stimulator for PG cells in vivo as well as in vitro.

Objective. To investigate the expression of telomerase gene hTRT mRNA in HeLa cells and to obtain hTRT pro-tein for futher study. Methods. The gene for encoding hTRT catalytic domain was cloned based on RT-PCR amplification from HeLa cells and sequenced. The cloned hTRTcDNA was in-frame inserted into His-tag fusion expression vector pEK318. The His-tag hTRT fusion proteins were purified by Ni-NTA chromatography and stained by westerm blotting. Results. An approximately 620bp fragment was generated and cloned into pBluescript SK + between Sail and BamHI sites. DNA sequencing showed the isolated fragment was consistem to those reported. SDS-PAGE present that a 17kDa protein was expressed stably in E. coli JM109 harboring pEKTRTM4 containing 6 × His-tag and hTRT 150aa, and the expression level of the protein was about 26% of the total bacterial proteins, while the expression of pEKTRT containing 6 × His-tag and hTRT 243aa was only detectable as 27 kDa band in western blotting. Both of fu-sion proteins were purified by Ni-NTA chromatography and showed single band( ＞ 95% purifity) in Coomassie Bril-liant staining. Westem-blotting confirmed that two proteins could be recognized by the Ni-NTA AP conjugate. Conclusions. The hTRT catalytic domain was highly conserved. The expressed hTRT protein contained recogniz-able His-tag, telomerase-specific and strong antigenic epitops, which may be convenient for further investigation.

Objective　To investigate genes involved in cancer metastasis, mRNA differential display was used to compare the levels of gene expression in two cell sublines derived from human giant cell carcinoma of lung (PG) which had different metastatic potentials. Methods　Using in vivo tumorigenicity and a spontaneous metastasis assay in nude mice, two sublines (BE1, LH7) from human giant cell carcinoma of lung (PG) with different metastatic potentials were isolated and characterized. mRNA differential display was used to compare the levels of gene expression between them and the obtained results were confirmed by Northern hybridization. Results　One differentially expressed band was nearly identical (99% homology) to Ras-GTPase-Activating protein SH3 domain binding protein (G3BP). G3BP displayed a strong expression in LH7 (non-metastatic in recipient nude mice) and a very weak expression in BE1 (100% metastatic frequency). The same different expression level of G3BP was detected in Northern hybridization with another panel of cell sublines with different metastatic potentials (established in our lab) derived from human prostate carcinoma cell line PC-3M. Conclusion　Our results indicate that G3BP was implicated in cancer metastasis because of its differential expressions in the two panels of cell sublines with different metastatic potentials.

Radiographic detection of pulmonary nodules based on three-dimensional Hessian matrix is highly sensitive but frequently produces false positive results in areas where blood vessels intersect. We propose a novel approach to pulmonary nodule detection using Hessian matrix-based adaptive window structure analysis, in which the structure coefficients is used to differentiate a voxel that belongs to a nodule or vascular structures, followed by construction of the 3D adaptive window to analyze the local structure characteristics; the nodules were then detected using the discrimination function. The experimental results on pulmonary CT images from 17 patients showed a 100% detection sensitivity for nodules of varying sizes and types, with also significantly reduced false positive results generated by the vessel junctions. This approach provides valuable assistance to follow-up positioning and segmentation of the pulmonary nodules.
Humans ; Lung ; pathology ; Lung Neoplasms ; diagnosis ; Tomography, X-Ray Computed

OBJECTIVETo investigate expression of Her-2/neu (c-erbB-2) and its significance in ovarian epithelial tumors.

METHODS106 specimens and clinical history of ovarian epithelial tumors were collected including 54 cases of malignant, 33 cases of borderline and 19 benign cases. There were 19 cases of stages I and II and 35 cases of stages III and IV in ovarian carcinoma and all borderline malignant cases were stage I according to FIGO's standard. Immunohistochemical staining for Her-2/neu was performed by LSAB method.

RESULTSThe positives rates of Her-2/neu of benign, borderline and malignant tumors were 47.4% (9/19), 84.8% (28/33,), and 85.2% (46/54), respectively. Both borderline and malignant cases were found to have higher expression of Her-2/neu than those of benign cases (P < 0.02 and P < 0.01) respectively. Overexpression of Her-2/neu showed in 14/54 (25.9%) of malignant cases, 3/33 (9.1%) of borderline, and none of benign cases. Overexpression of Her-2/neu in malignant tumors cases with metastasis was higher than those without metastasis (P < 0.001).

CONCLUSIONOverexpression of Her-2/neu is associated with biological aggressiveness of ovarian cancer.

OBJECTIVETo investigate the pathological course in intracranial aneurysms.

METHODSNormal intracranial artery tissue (cortex fistulization) from 1 case, ruptured aneurysms tissuses from 11 cases, unruptured aneurysm tissues from 2 cases were obtained by neurosurgical excision. Routine HE staining was used to observe histological characteristics. In situ hybridization was used to observe the expression of the monocyte chemoattractant protein-1 (MCP-1) mRNA in the walls of the normal artery and aneurysms.

RESULTSBy the HE staining showed that the wall of the ruptured aneurysms (10 cases) and unruptured ones (2 cases) had increased intima and connectivum extima. The fibroblast in the intima was arrayed in the disorder. Monocyte-like cells can be seen in the whole aneurysm wall. In one case aneurysms wall (ruptured) glass-like fiber structure was left over, few cells could be seen. In 9 cases, mural thrombus was found. The thrombus represented with organization. In situ hybridization, MCP-1 mRNA was not detectable in the normal artery. The hybridization signal could be observed in the ruptured aneurysms (10 cases) and unruptured ones (2 cases) often in the intima. MCP-1 mRNA appeared to be expressed by fibroblast cells in its cytoplasm. Monocyte-like cells had little cytoplasm, and the signal was seldom seen. The hybridization signal was discontinuous in the intima, MCP-1 mRNA expressed where fibroblast and monocyte-like cells assembled. One ruptured aneurysm had no signal because there were no cells only glass-like fiber. Mural thrombus showed upregulated hybridization signal in the cytoplasm of fibroblasts, phlogocytes and endotheliocytes of its micrangium.

CONCLUSIONThe pathological representation of the ruptured and unruptured aneurysms and the upregulated expresion of MCP-1 in the aneurysm wall suggest that the development of aneurysm may be a course of chronic inflammation in which main inflammatory cells are monocyte-like cells.