Objective To establish a luciferase labeled McA-RH7777 hepatoma rat model,which could be used for gross observation to further observe the effect of selective ligation of the portal vein and bile duct on tumor growth and metastasis.Methods The luciferase gene was transfected into rat McA-RH7777 hepatoma cells with pCDH-puromycin-CMV as the carrier,which were subcutaneously inoculated into Buffalo rats.Tumor pieces were then heterotransplanted into the left lateral lobe of the allogenic rat liver to observe the tumor growth in vivo.After the successful hepatoma modeling,the rats were randomly divided into three groups,namely the implanted portal vein group with combined portal vein and bile duct ligation,the implanted portal vein group with single portal vein ligation and sham operation group.The rats were executed at the 1 st week and 2nd week after ligation,and the livers were dissected to record the tumor growth and metastasis inside and outside the liver,respectively.Results The tumor formation rates of Buffalo rats after subcutaneous and intrahepatic implantation were both 100％.The fluorescence signal implanted into the liver lobe could be observed in vivo after the intrahepatic implantation of luciferase transfected Luc-McA-RH7777 at 2nd week,the range and intensity of which increased over time.Only local tumor growth could be found at the 4th week,without obvious intrahepatic and lung metastasis.However,both an increased in situ tumor volume and the pulmonary metastasis could be observed in the implanted portal vein group with combined portal vein and bile duct ligation at 2nd week after the ligation.Immunohistochemistry showed AFP positive immunoreactions in the vast majority of intrahepatic tumor cells and Luc positive immunoreactions in part of tumor cells.Conclusion Luc-McA-RH7777 cells could be used to establish the heptoma rat model and the in vivo analysis within the Buffalo rat liver demonstrated that the combined ligation of the portal vein and bile duct can accelerate the development and metastasis of liver cancer.

Adult ; Epilepsy ; chemically induced ; therapy ; Humans ; Male ; Nitrobenzenes ; poisoning ; Occupational Exposure ; Poisoning ; complications ; therapy

Aim: Clone and characterize of the 5′- flanking region of the nitrate reductase (NR) gene derived from Dunaliella salina(D. salina). Methods : The genomic DNA from D. salina was respectively digested with BamHI, EcoRI, HindIII, Pst I, Sal I and Xba I. A genomic walking cassette was ligated to the ends of the digested DNA fragments, and then genomic walking libraries comprising BL, EL, HL, PL, SL and XL were constsucted. The 5′- flanking region of the NR gene from genomic walking libraries of D. salina was amplified by LA-PCR. The DNA sequences were analyzed with the software - Promoter Predictions. Isolated 5′-flanking regions fused to the GUS gene were tested for transient expression in the alga. Results: A single specific PCR product of about 1200bp in length from the HL library was generated. Also, several conserved motifs, such as CAAT-box, GAGA-box were found, which are related to regulation of transcription, and the putative binding sites of transcriptional factors such as EBP, EFII, NF-E1 and LV. BLAST showed that the DNA sequences shared high homology with 5′-upstream region of the NR gene from Dunaliella viridis. The isolated 5′-flanking regions were able to strongly drive GUS reporter gene expression, suggesting that it contains the promoter elements necessary for the transcription of the NR gene. The expression pattern of the GUS gene and the NR gene were similar, both ware induced by nitrate and repressed by ammonium. Conclusion: The cloned 5′- flanking sequences of NR gene derived from D. salina might be a specific promoter with the ability to“switch on or off” an expression of the heterologous gene in transgenic D. salina.

Solitary fibroma is a rare spindle cell tumor.This article reports a case of bladder solitary fibroma to introduce its clinical diagnosis and treatment.The patient,male,43 years old,was admitted to the hospital because of "discovery of gross hematuria for 2 days".CT scan suggested the space-occupying lesion of the bladder area.Pathological results showed the spindle cell tumor.Further immunohistochemistry suggested lowgrade malignant solitary fiber tumor.

OBJECTIVETo use a new kind of fixing material, i.e. Sol-Gel organic-inorganic hybridized material to immobilize bacterium to detect Biochemical oxygen demand quickly.

METHODSThe biosensor was fabricated using a thin film in which Hansenula anomala was immobilized by sol-gel and an oxygen electrode. The optimum measurement for biochemical oxygen demand was at pH 7.0; 28 degrees C; response time 3 - 12 min. Pure organic compound, sewage and rate of recovery were detected with the biosensor.

RESULTSIt shows that the BOD biosensor can be used to detect many organic compounds such as amino acid, glucide. It is suitable to monitor sewage and industrial waste water which has low level alcohols and phenols. The microbial membrane can work 3 months and remain its 70% activity. It is measured that the rate of recovery of BOD is between 90% to 105% in sewage.

CONCLUSIONThe study confirmed the effectiveness and usefulness of BOD sensor, which is quick, convenient, low cost and reliable with little interference.

Bacteria ; Biosensing Techniques ; instrumentation ; Cells, Immobilized ; Gels ; Membranes, Artificial ; Nylons ; Oxygen ; analysis ; Sewage ; analysis ; microbiology

ZM-1 tissue microarrayer designed by our group is manufactured in stainless steel and brass. It features an easier and faster preparation for tissue microarrays. By means of it, a group of biopsy needles are used to punch the donor tissue specimens respectively, and all the needles with the punched specimen cylinders are arranged into the array-board, where small holes have been digged to fit the needles. All the specimen cylinders arraying and the tissue microarray block's shaping are finished simultaneously. ZM-1 tissue microarrayer with a lower cost of manufacture, is capable of preparing the tissue microarrays conveniently, efficiently and quality-controllably.
Equipment Design ; Tissue Array Analysis ; instrumentation

OBJECTIVETo investigate the effects of microencapsulated Chinese hamster ovary (CHO) cells modified with maspin gene on the motility and adhesiveness of breast carcinoma cells Bcap37 and to explore the possibility and feasibility of its clinical application in treatment of malignant tumors.

METHODSAfter the Bcap37 cells were co-cultured with the microencapsulated CHO cells modified with maspin gene, their motility and adhesion to vascular endothelial cells (ECV304), changes in CD44v6 and E-cadherin expression were examined.

RESULTSAfter the treatment, the motility of Bcap37 cells, their adhesion to vascular endothelial cells ECV304 and the CD44v6 expression were significantly reduced. The adhesiveness of Bcap37 cells and their E-cadherin expression were significantly enhanced.

CONCLUSIONThe microencapsulated CHO cells modified with maspin gene decrease motility and adhesiveness of breast carcinoma cells Bcap37, which help explain the anti-metastatic effects of maspin.

Animals ; Breast Neoplasms ; pathology ; CHO Cells ; Capsules ; Cell Adhesion ; Cell Movement ; Coculture Techniques ; Cricetinae ; Cricetulus ; Female ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Hyaluronan Receptors ; biosynthesis ; genetics ; Serpins ; genetics ; Tumor Cells, Cultured

The ZM-1 tissue microarrayer designed by our groups is manufactured in stainless steel and brass and contains many features that make TMA (tissue microarray) paraffin blocks construction faster and more convenient. By means of ZM-1 tissue microarrayer, biopsy needles are used to punch the donor tissue specimens respectively. All the needles with the punched specimen cylinders are arrayed into the array-board, with an array of small holes dug to fit the needles. All the specimen cylinders arraying and the TMA paraffin block shaping are finished in only one step so that the specimen cylinders and the paraffin of the TMA block can very easily be incorporated and the recipient paraffin blocks need not be made in advance, and the paraffin used is the same as that for conventional pathology purpose. ZM-1 tissue microarrayer is easy to be manufactured, does not need any precision location system, and so is much cheaper than the currently used instrument. Our method's relatively cheap and simple ZM-1 tissue microarrayer technique of constructing TMA paraffin block may facilitate popularization of the TMA technology.
Biopsy, Needle ; instrumentation ; Equipment Design ; Female ; Humans ; Immunohistochemistry ; Male ; Neoplasms ; enzymology ; Paraffin ; Tissue Array Analysis ; instrumentation

OBJECTIVETo construct a recombinant adenovirus expression vector containing the anti-oncogene PTEN and to investigate the effects of the PTEN gene on the proliferation of prostate cancer PC-3 cells and the expressions of cyclin D1 and p21 in the PC-3 cells.

METHODSThe PTEN gene was amplified from the rat hippocampus by RT-PCR and cloned into the shuttle plasmid pEN-TR2A. The plasmids were constructed and amplified in 293A cells. Prostate cancer PC-3 cells were cultured in vitro and infected with the adenoviral vector carrying the PTEN gene (Ad-PTEN). The up-regulation of the PTEN protein was measured by indirect immuno-fluorescence assay; the expressions of PTEN, cyclin D1 and p21 in the cells infected with Ad-PTEN and Ad-LacZ were determined by

RESULTSThe Western blot; and the effect of PTEN on the cell proliferation was detected by MTT assay and plate colony formation. recombinant adenoviral vector Ad-PTEN was successfully constructed. Western blot showed a significantly increased expression of the PTEN protein in the PC-3 cells infected with Ad-PTIEN (0.215 +/-0.065) as compared with that in the control ([0.052 +/-0.009], t = 4. 30, P <0.05) and the Ad-LacZ group ( [0. 056 +/- 0.008 ] , t =4.21, P <0.05). The expression of cyclin D1 was significantly lower in the Ad-PTEN-infected PC-3 cells (0. 256 +/- 0. 072) than in the control ( [0. 502 +/- 0. 087 ], t = 3.77, P < 0.05) and the Ad-LacZ group ([0.498 +/-0.081] , t =3.87, P <0.05), while the expression of p21 remarkably higher in the Ad-PTEN-infected PC-3 cells (0.589 +/-0. 076) than in the control ([0. 146 +/-0.026] , t = 9.55, P<0. 01) and the Ad-LacZ group ([0. 163 +/-0. 024] , t = 9.26, P <0.01). Ad-PTEN significantly inhibited the growth of the PC-3 cells (21.98%) at 48 h (t = 6.80, P <0.01). The colony formation rate of the PC-3 cells was (37.4 +/-4. 18)% in the Ad-PTEN group, significantly lower than (54.9 +/-4.81)% in the control (t =4.76, P<0.01) and (56.5 +/- 5.42)% in the Ad-LacZ group (t=4.83, P<0.01).

CONCLUSIONThe expression of PTEN induced by Ad-PTEN can significantly inhibit the proliferation of PC-3 cells, down-regulate the expression of cyclin D1, and up-regulate the expression of p21.

Adenoviridae ; genetics ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Humans ; Male ; PTEN Phosphohydrolase ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo prepare traditional Chinese medicine formula Qiqi pellets by extrusion-spheronization and study the optimal formulation and process.

METHODQiqi pellets were prepared by a new style extrusion-spheronization equipment, the optimal formulation and process was obtained by the studies of influenitial factors and L9 (3(4)) orthogonal design, the micromeritic properties and product yield of pellets were determined.

RESULTThe formula Qiqi pellets prepared by extrusion-spheronization were all spheral with smooth surface; the product yield was high.

CONCLUSIONExtrusion-spheronization was suitable to produce Chinese Herbal Medicine pellets. The preparation process was simple and feasible; The quality of the prepared pellets was excellent. It was worth further study.

Astragalus membranaceus ; chemistry ; Cellulose ; chemistry ; Drug Combinations ; Drug Compounding ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethanol ; chemistry ; Lycium ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; instrumentation ; methods ; Water ; chemistry