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 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

AIMTo study the chemical composition of Opuntia dillenii Haw.

METHODSMany kinds of chromatography methods were used to separate the chemical constituents. Their structures were determined by NMR and MS spectral analysis.

RESULTSA new compound, together with five known compounds, were isolated from the 80% ethanolic extract of the stems.

CONCLUSIONThe new compound was identified as 4-ethoxyl-6-hydroxymethyl-alpha-pyrone. Compounds 1, 3, 4 and 5 were obtained for the first time from the genus of Opuntia, and they were: 3-O-methyl isorhamnein, 1-heptanecanol, vanillic acid, isorhamnetin-3-O-beta-D-rutinoside. Ruin was isolated from this plant for the first time.

Molecular Structure ; Opuntia ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Pyrones ; chemistry ; isolation & purification ; Rutin ; chemistry ; isolation & purification ; Vanillic Acid ; chemistry ; isolation & purification

Objective To investigate the expression and correlation of NKG2D and sMICA in lung cancer patients. Methods By collecting 30 lung cancer patients as the test group,and taking 30 healthy volunteers as the contrast group, the expression of NKG2D and sMICA in the two groups were examined separately by FACS and ELISA method. Results The expressions of NKG2D in the two groups were (81.56±8.78) %, (85.63±6.62) %. The lung cancer patients were high remarkable. There was a significant difference between the two groups (P <0.05). The expression of sMICA in the two groups were (354.13 ±80.575) pg/ml,(216.53±48.175) pg/ml. The lung cancer patients were low remarkable. There was a significant difference between the two groups (P <0.01). There was a significant relation between the two groups (r =-0.349, P =0.006). Conclusion The expression of NKG2D and sMICA may provid one of the immune targets for diagnosing that can forecast the immune state and malignant metastasis of the lung cancer patients. The significant relation between NKG2D and sMICA may take on main role in the immune escaping of tumor. It may provide the suitable target of the patients for tumor organisms and immune treatment.