Fusion of human orphan G protein-coupled receptors GPR45, GPR85 or GPR174 with Gi1α and their expression in insect Sf9 cells / 军事医学科学院院刊

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Laparoscopy-assisted D2 total gastrectomy in advanced gastric cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and advantages of laparoscopy-assisted total gastrectomy (LATG) with D2 dissection of lymph nodes versus conventional open D2 total gastrectomy (OTG) in advanced gastric cancer.</p><p><b>METHODS</b>One hundred and twenty-five patients with advanced gastric cancer in the middle or upper third of the stomach were operated on from July 2005 to March 2007. Of the patients, 59 cases received LATG and 66 OTG with D2 lymph nodes dissection. Clinical data were recorded and compared between the two groups.</p><p><b>RESULTS</b>No patient in the LATG group converted to conventional operation with laparotomy. No operation mortality and no severe morbidity occurred in LATG group. As compared with OTG group, in LATG group operation time was longer [(330 +/- 71) min vs. (261 +/- 54) min, P =0.005] in LATG group, but with similar number of lymph node retrieval (36 +/- 13 vs. 34 +/- 16, P =0.450), less operation blood loss [(175 +/- 101) ml vs. (359 +/- 210) ml, P =0.003], earlier recovery of bowel activity (P = 0.015), and a shorter duration of fever after operation (P = 0.024).</p><p><b>CONCLUSIONS</b>LATG with D2 lymph node dissection in advanced gastric cancer is safe and technically feasible with better operative access and visual field, less operation blood loss and earlier recovery.</p>

Isolation of a down-regulated novel gene with lower abundance in gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To clone novel gene from suppression subtraction library established for screening down-regulated genes in gastric carcinoma, and the effects of novel gene on gastric tumorigenicity were analyzed.</p><p><b>METHODS</b>Sequencing results of 860 positive colonies chosen randomly were compared by Blast program in GenBank. Novel gene fragment was amplified by rapid amplification of cDNA ends (RACE). The mRNA expression of novel gene was detected by Northern blot and semi-quantitative PCR in 25 cases of gastric carcinoma tissue and counterpart normal gastric mucosa. The structure and chromosomal location of novel gene were investigated by Bio-message technique.</p><p><b>RESULTS</b>A 233 bp novel gene fragment was screened out from 860 clones and a 802 bp novel gene was obtained by RACE. The novel gene was named as GDDM, registered in the number of AF494508 by GenBank. The mRNA expression of GDDM in gastric carcinoma tissue (4.496+/-0.637) was significantly lower than that in the counterpart normal gastric mucosa (36.919+/-6.290)(P<0.01). Chromosomal location of GDDM gene was at 4q31.</p><p><b>CONCLUSION</b>The cloned novel gene, GDDM, is down-regulated in gastric carcinoma, and it is likely to be involved in gastric tumorigenicity.</p>

Lycopene liposomes: lycopene release in vitro and pharmaceutical behaviors and antioxidation in vivo / 药学学报

Lycopene liposomes were prepared by conventional rotary-evaporated film-ultrasonication method. The release of lycopene from lycopene liposome was evaluated in vitro. The pharmacokinetic parameters of lycopene liposomes (L-LYC) and lycopene (LYC) oil, the effect of LYC and L-LYC on antioxidation were also investigated in rats. HPLC method was used to assay the concentration of lycopene in rat's plasma. Pharmacokinetic parameters were estimated by 3P97 program. The release of L-LYC and LYC were measured in the artificial stomach liquid and bowel liquid. After 4 weeks of L-LYC or LYC feeding, the activity of SOD, T-AOC, GSH-Px, MDA and CAT in serum and liver were measured separately. The pharmacokinetic parameters of LYC oil and L-LYC in a single dose were 4.45 and 7.45 h for Tmax; 0.473 and 0.654 microg x mL(-1) for Cmax; 12.38 and 21.67 mirog x h x mL(-1) for AUC,respectively. The activities of GSH-Px and T-AOC in serum and liver of the L-LYC group increased (P < 0.05) and the concentrations of MDA and CAT decreased significantly (P < 0.05). It could be concluded that lycopene liposomes could prolong the time of absorption. L-LYC could increase antioxidative effect and reduce lipid peroxidation obviously compared with LYC in rats.

Study on novel gene GDDR related to gastric cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To confirm the GDDR cDNA property of novel down-regulated full-length gene in gastric cancer, structure of genomic GDDR DNA and its promotor region. To predict its transcription factors and transcription factor binding sites. To explore function of GDDR gene in vitro.</p><p><b>METHODS</b>GDDR mRNA was located by in situ mRNA hybridization of gastric mucous membranes, and was amplified in 13 human organs and tissues. The structure and location of GDDR on chromosome, property of protein encoded by full-length GDDR were investigated by Bio-message technique. Promotor region of GDDR was confirmed, and transcription factors or their binding sites were predicted in software Gene2promoter and Matinspector of Genomatix. Both of vector pcDNA3.1/Myc-His(-)A inserted by GDDR ORF and control vector pcDNA3.1/Myc-His(-)A were respectively transfected into gastric cell lines 7901 by lipofectamin. Growth curve, MTT test and a morphological analysis were respectively performed.</p><p><b>RESULTS</b>GDDR mRNA was located in gastric mucous epithelial cells, and only was expressed in gastric tissue. 7739 bp genomic GDDR DNA located on chromosome 2p13.3, 21701 bp away from CA11-one stomach-specific gene related to gastric cancer. 618 bp promotor region of GDDR located at position +96 bp,and -419 bp of transcription start site of GDDR. The structure of genomic DNA or cDNA between gene GDDR and CA11 was mostly similar. Sequences of their promotor region were different, transcription factors and their binding sites also varied between gene GDDR and CA11. GDDR encoded protein including a trans-membrane peptide homologed to CA11 that have been proven to encode secrete protein. GDDR was another new member of BRCHOS family just was found. Gastric cell lines 7901 transfected by GDDR showed a marked decrease in growth rate by growth curve and MTT test (72 h, 0.341 +/- 0.014 vs 0.488 +/- 0.015 A, P < 0.01).</p><p><b>CONCLUSIONS</b>Stamoch-specific, novel down-regulated gene GDDR in gastric cancer locates in gastric mucous epithelial cells can markedly inhibit growth of gastric cancer cell lines 7901, GDDR is another new member of BRICHOS family related to gastric cancer except CA11.</p>

Researches on the establishment of chronic nephropathy models and the effect of TCM on these models / 中国中西医结合杂志

Following contents were reviewed in this article: More and more experimental studies related with chronic uratic nephropathy were carried out in recent years. In most of these studies, the animal models were established mainly from viewpoints of gene recombination, urinary uric acid inhibition and blood uric acid production promoting. TCM showed good effects in lowering blood uric acid, regulating levels of cytokines and postponing interstitial fibrosis. However, further studies on Chinese herbs and their extracts is necessary.

The classification and management of pancreatic duct stone / 中华外科杂志

<p><b>OBJECTIVE</b>To set up a stand for surgical classification of pancreatic duct stone and evaluate the benefits of different management according to the classification.</p><p><b>METHODS</b>Retrospectively analysis the diagnosis and prognosis of different management of 33 cases pancreatic duct stones to establish a new standard of classification and strategy of management of pancreatic duct stone.</p><p><b>RESULTS</b>According to the results of imaging examination (B-US, CT, ERCP) and finding during surgery, pancreatic duct stone can be classified into four different types: Type I: The stones mainly located in the head of pancreas. Endoscopic pancreas drainage and remove of stones is the first line choice of treatment. If it fail the Whipple procedure should be applied. Type II, The stones mainly located in the body of pancreas. It can be treated by Pusetow procedure. Type III, The stones mainly located in the tail of pancreas. The resection of the tail of pancreas or combined with spleenectomy was recommended for the management of this type stones. Type IV, The stones can be found from the head to tail of the main duct of pancreas. The Pusetow-Gillesby procedure or dividing of the neck of pancreas removing stones from both ends of pancreatic duct and reconstructed by two ends pancreatic duct-ileostomy in Roux-en-Y fashion are the choice of management.</p><p><b>CONCLUSION</b>The invadulaized strategy of the management based upon correct diagnosis and classification play the most important role in the treatment of pancreatic duct stone.</p>