To developing a simple, rapid and sensitive immunoaffinity method for extracting DNA before amplification by the polymerase chain reaction(PCR). Using monoclonal anti-DNA bound to colloidal-gold beads(which called immunocolloidal-gold ) and extracting DNA from the specimen before amplification by PCR. The immunoaffinity method we use is extremely efficient in extracting DNA at low concentration, also, the beads carrying the DNA/anti-DNA complexes can be added directly to a PCR mixture without elution, and the presence of PCR inhibitors in specimens has no effect on amplification. When applied to serial dilution bacteria suspension, the detection of limit can be 100 CFU/mL, and the sensitivity of detecting artificial soil sample and milk sample were 101 and 100 CFU/mL. The immunoaffinity method described here can sever as a sensitive and practicable method for extracting DNA , particularly where samples have low concentrations of DNA or where the material is in poor condition.

Objective To analyze the manifestation,reason,the processing method of the steel wire implantation with the sereotactic mammography to improve the accuracy of the preoperative positioning.Methods Seventy-nine cases which got the stereotactic steel wire implantation.In 96 lesions, 13 had steel wire displacement.Among them,5 cases got steel wire displacement during the sereotactic process,5 cases got steel wire displacement after the stereotactic process,2 cases got steel wire displacement during the operation,one case did not show the calcification on the postoperative radiography.Results The steel wire displacement occurred in 5 cases during the stereotactic process came from the patients and doctors respectively and the repositioning was needed.The steel wire displacement after the stereoscopic positioning was attributed to the overdose injection of local anesthesia,which led to the mismatch between the depth of Z axis of the mammary gland and the actual depth the computer given,the incorrect method for needle placement,and,neglecting whether the steel wire have got the lesion anchored when pulling out the needle set of steel wire hood,besides,these three kinds of instances above were all exaggerated by the accordion effect.For the displacement within 2 cm,the lesion can be excised toward the pathological change direction according to the position that steel wire prompted and re-place the second steel wire,putting the J-shaped steel wire into the needle hood and taking it out of the body.After repositioning,2 cases had the steel wire prolapse during operation,which resulted from the over-lifting of the steel wire.After placing the steel wire, the radiologist should give an accurate description on the depth and direction to the surgeon and the notch should be taken for incision from the steel wire head end which is proximate to skin.The postoperative specimen from one case had no calcification,which might be related to the condition that the calcification was located in the gland body,which got destruction from the surgical electrical electrotome.The excisionscope should be extended and the short term reexamination is recommended to make sure the complete excision of the calcification.Conclusion It is the gold standard method that implanting the steel wire with the stereotactic mammography to guide the surgical dissecting technique to diagnose non-palpable breast lesion(NPBL).Thorough understanding of the displacement manifestation of implanting steel wire with stereotactic technique and the treatment methods will be helpful in the surgical dissecting guidance.

Alkaloids ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Anticonvulsants ; pharmacology ; Benzodioxoles ; Oils, Volatile ; isolation & purification ; pharmacology ; Piper ; chemistry ; Piper nigrum ; chemistry ; Piperidines ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Polyunsaturated Alkamides

Objective To investigate the expression of inregrin?_3 and integrin?_1 in breast cancer and its bio- logical significance.Methods Immunohistochemical assay was used to determine the expression of integrin?_3 and integrin?_1 in the breast cancer(32 cases).Results In normal breast tissue,the positive expression rates of integrin?_3 and integrin?_1 were 0 % and 25 %.In the breast cancer tissue,the positive expression rates of integrin?_3 and inte- grin?_1 were 36 % and 81%.Conclusion The integrin?_3 and integrin?_1 are close associated with the biological sig- nificance of breast cancer.To examine its expression is useful to evaluate the aggressive degree,metastatic potential and prognosis in patients with breast cancer.

OBJECTIVETo isolate and elucidate the constituents from the roots of Commercial Ginseng.

METHODColumn chromatography and HPLC were used to isolate chemical constituents. Physico-chemical characters and spectr-oscopic analysis were employed for structural identification.

RESULTSixteen compounds were identified as: notoginsenoside-R2(1), ginsenoside-Rg2(2), 20 (R)-Rg2 (3), ginsenoside-Rg1 (4), -Rf(5), -Re(6), -Rd(7), -Rc(8), -Rb1(9), -Rb2(10), -Rb3(11), -Ra3(12), -Ra2(13), -Ra1 (14), notoginsenoside-R4(15) and ginsenoside -Ro(16).

CONCLUSIONCompound 1 was obtained from the plant for the first time.

Ginsenosides ; chemistry ; isolation & purification ; Panax ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Objective To investigate the effect of OX40/OX40L interaction on the nuclear factor of activated T cells c1(NFATc1) in ApoE-/- mice.Methods Lymphocytes were prepared from mouse spleens after Collar-treated Surgery, then incubated with a range of agonistic anti-OX40 mAbs and inhibitory anti-OX40L mAb to stimulate or inhibit OX40-OX40L interaction in vitro. The expression of NFATc1 mRNA and protein in lymphocytes of ApoE-/- mice was measured by Real Time PCR and flow cytometry, respectively.Results (1)After stimulating OX40-OX40L signal pathway, the expression of NFATc1 mRNA and protein in leukocytes of ApoE-/- mice was significantly increased, with maximal effect occurring at 20 μg/ml anti-OX40 mAb- stimulated, and peaked at 24 h at any concentration(P<0.01). (2)Anti-OX40L mAb significantly suppressed the expression of NFATc1 in leukocytes of ApoE-/- mice, with maximal effect occurring at 20 μg/ml anti-OX40L mAb, and peaked at 24 h(P<0.001).ConclusionsOX40-OX40L interaction can regulate the mRNA and protein expressions of NFATc1 in lymphocytes of ApoE-/- mice.

OBJECTIVEIn order to clarify the exact molecular weight of tumor metastasis suppressor gene-1 (TMSG-1) protein and its cellular localization, a monoclonal antibody against TMSG-1 was prepared, characterized and applied to evaluate the metastatic potential of human tumors.

METHODSA dominant epitope-TMSG-1(15)-derived from TMSG-1 was synthesized based on Fmoc method, and the hapten was conjugated to Imject Maleimide activated mcKLH as a carrier protein. The antigen preparation was used to immunize BAL B/C mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, which were further characterized by western blotting and immunohistochemical staining.

RESULTSOne hybridoma cell line secreting anti-TMSG-1 antibody, designated as C8, was eventually established after primary ELISA screening, followed by rapid limited dilution procedure. It was confirmed that C8 was of IgM isotype. Result of competitive inhibition assay showed that the antibody was TMSG-1 specific. Using this antibody, an expected protein band of about 45,000 (relative molecular mass) was detected in the non-metastatic variants PC(3)-2B4 and PG-LH7 cells by Western blotting, but not in the isogenetic metastatic variants of PC3-1E8 and PG-BE1 cells. Immunohistochemistry using C8 showed a positive staining of cell membrane and cytoplasm of 2B4 and LH7 cells, whereas 1E8 and BE1 cells were non-reactive. Immunostaining using C8 of paraffin sections of 52 breast carcinomas and 41 colon cancers demonstrated a strong positivity in non-metastatic tumors, but none to weakly reactive in metastatic tumors (P < 0.05).

CONCLUSIONC8 monoclonal antibody against the synthetic peptide is TMSG-1 specific and is effective for Western blot and immunohistochemistry assays to detect TMSG-1 expression in cancer cells. TMSG-1 protein is about 45 000 (relative molecular mass) at cell membrane and cytoplasm of tumor cells. Expression of TMSG-1 protein correlates well, inversely with the tumor metastatic potential.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Cytoplasm ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hybridomas ; immunology ; secretion ; Male ; Membrane Proteins ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Neoplasm Metastasis ; Sphingosine N-Acyltransferase ; Tumor Suppressor Proteins ; immunology ; metabolism

OBJECTIVETo investigate the clinical effect of atlantoaxial pedicle screw internal fixation for treatment of atlantoaxial dislocation.

METHODSSixteen patients with atlantoaxial dislocation were treated from Dec. 2005 to June 2007, included 10 males and 6 females, aged from 38 to 45 years old (means 40.5 years). Among them 12 patients combined with nerve injury, according to ASIA grade: there were 3 cases in grade B, 5 cases in grade C, 4 cases in grade D. All patients received preoperative CT, radiograph and skull traction. Intraoperative posterior approach general spine pedicle screw-rod orthopaedics fixation system used and iliac bone block were implanted in space of posterior atlantal arch and axial vertebral plate. The outcome and complications were observed in the near future.

RESULTSThere was no vascular or neural injury found. The patients were followed up for 12 to 24 months (means 18 months). All head pain, acid storm symptoms were improved after operation. According to the Odom's clinical efficacy evaluation standard, 12 cases were excellent, 4 were good. Eleven cases of 12 with nerve injury recovered significantly. By ASIA classification: 1 cases was in grade B, 2 cases were in grade C, 5 cases were in grade D, 4 cases were in grade E. No looseness or breakage of screw occurred. Bony fusion was achieved in all cases.

CONCLUSIONPosterior approach atlantoaxial pedicle screw internal fixation have the advantages of direct screw placement, short-segment fusion, intraoperative reduction, fixation reliable, high fusion rate, and it can restablish the upper cervical vertebrae stability and help to recover the spinal cord and nerve function.

Adult ; Atlanto-Axial Joint ; diagnostic imaging ; injuries ; physiopathology ; surgery ; Bone Screws ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Joint Dislocations ; diagnostic imaging ; physiopathology ; surgery ; therapy ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo investigate the frequency of EML4-ALK fusion gene in non-small-cell lung cancer (NSCLC) patients, and its correlation with clinicopathologic features.

METHODSReal-time PCR was used to detect the presence of EML4-ALK fusion gene in 268 cases of NSCLCs using paraffin-embedded tissue samples(among which 164 samples were re-validated by Sanger sequencing). Related clinicopathological correlation was analyzed.

RESULTSEML4-ALK fusion gene was found in 4.1% (11/268) of the cases. One hundred and sixty four samples were verified by Sanger sequencing, and the overall coincidence of the results of two methods (Sanger sequencing and Real-time PCR) was 100%. Female patients (5.9%, 5/85), ≤ 60 years of age (4.3%, 6/140), non-smokers (6.8%, 8/118) and adenocarcinomas (7.6%, 10/132) had a higher mutation rate than that in male patients (3.3%, 6/183), > 60 years of age (4.0%, 5/124), smokers (1.6%, 2/132) and squamous cell carcinomas (1.3%, 1/79), although no statistical significance in age (P = 0.918), gender (P = 0.503), smoking history (P = 0.092) and histological type (P = 0.094).

CONCLUSIONSChinese NSCLC patients have a 4.1% detection rate of EML4-ALK fusion gene in the tumor tissues. Female, non-smoker and adenocarcinoma histological subtype tend to be associated with a higher rate of EML4-ALK gene fusion.

Adenocarcinoma ; genetics ; metabolism ; pathology ; surgery ; Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; surgery ; Female ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; metabolism ; Sex Factors ; Smoking ; Young Adult

OBJECTIVETo study the histopathologic features, differential diagnosis and prognosis of epithelioid angiomyolipoma (EAML) of kidney.

METHODSTwo cases of EAML (including one case with recurrence) of kidney were retrieved from the archival files of Departments of Pathology, Navy General Hospital of PLA and Health Science Center of Peking University. The clinicopathologic features, immunohistochemistry, ultrastructural findings and follow-up data were studied and literature reviewed.

RESULTSHistologically, the tumors were predominantly composed of epithelioid cells with marked cellular pleomorphism. Focal perivascular arrangement was seen. Hemorrhage and necrosis were present and tumor emboli were found in the venous structures. The renal hilar lymph nodes were also involved by tumor cells. Immunohistochemical study showed that the tumor cells (including those in the hilar lymph nodes) were strongly and diffusely positive for HMB45, smooth muscle actin, neuron-specific enolase and vimentin. They were focally positive for S-100 protein, melan-pan and CD68. The staining for epithelial membrane antigen, AE1/3, CK7, CD117, muscle-specific actin, desmin, leukocyte common antigen, CD20, CD45RO, CD30, CD15, chromogranin A, synaptophysin, bcl-2, estrogen receptor, progesterone receptor and p53 were negative. Ultrastructural examination revealed the presence of melanosome-like dense granules, myofilaments and dense bodies in the tumor cell cytoplasm. Discontinuous and focally thickened basal lamina was seen surrounding the tumor cells. On follow up, both patients remained well and disease-free 10 months after operation.

CONCLUSIONSEAML is predominantly or almost entirely composed of epithelioid cells. Perivascular cellular arrangement, focal features of otherwise classic angiomyolipoma, as well as coexpression of HMB-45 and smooth muscle actin are clues to the correct diagnosis. The degree of cytologic atypia, presence of hemorrhage and necrosis and high mitotic activity may indicate the malignant potential of this tumor. On the other hand, the presence of lymph node involvement and even tumor emboli in renal veins may represent multifocaltumorigenicity rather than true malignancy. Definitive evidence of malignancy requires demonstration of distant metastasis.

Adult ; Angiomyolipoma ; metabolism ; pathology ; surgery ; Antigens, Neoplasm ; metabolism ; Diagnosis, Differential ; Epithelioid Cells ; pathology ; ultrastructure ; Humans ; Immunohistochemistry ; Kidney ; pathology ; ultrastructure ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Melanoma-Specific Antigens ; Microscopy, Electron ; Neoplasm Proteins ; metabolism