Objective To clone human bone morphogenetic protein 2 ( BMP-2) gene and construct the gene’s eukaryotic expression vector. Methods The total RNA was extracted from human osteosarcoma cells, the human BMP-2 cDNA was amplified by RT-PCR and inserted into pGEM-T vector. The positive clones were screened out, and the n the recombinant plasmid was confirmed by restriction enzyme digestion, PCR and the analysis of nucleotide sequence. The BMP-2 cDNA in the pGEM-T cloning vec tor was inserted into the pcDNA3.1(+) eukaryotic expression vector. Results The agarose electrophoresis showed that the fragments of BMP-2, pGEMT and pcDNA3.1(+) were 1.2 kbp, 4.0 kbp and 5.0 kbp, respectively. The result of nucleotide sequence confirmed that the cDNA sequence, which was inserted into pGEM-T and pcDNA3.1(+) plasmid was human BMP-2. Conclusion The pcDNA3.1(+)-hBMP-2 eukaryotic vector can be successfully constructed.
Objective To investigate the clinical significance of the level of serum amylase and serum IgA and total IgE in henoch-schonlein purpura patients with gastrointestinal involvement (also known as "Henoch purpura "). Methods Levels of serum amylase and serum IgA and total IgE in henoch-schonlein purpura patients with or without abdominal pain or patients with acute abdominal pain were compared. Results The average level (180.3 ± 15.8 IU) of serum amylase of Henoch purpura patients was significantly higher than HSP patients without abdominal pain and acute abdominal pain patients (F=32.214, P=0.009); Ratio of cases of increased level of serum IgA in henoch purpura abdomen patients was 44.2%, and there was no significant difference with HSP patients without abdominal pain. But ratio of two groups was respectively higher than the acute abdominal pain patients group (χ2=13.73, P=0.001); Ratio of cases of increased level of serum IgE in Henoch purpura abdomen patients accounted for 40.4%, but there was no significant difference among the three group (χ2=1.80，P=0.41). Conclusion Levels of serum amylase increase and serum IgA increase conduce to diagnose HSP patients with the onset of abdominal pain, and serum total IgE has little significance.
Objective To clarify the role of gastrokine 1 in the process of formation and development of gastric cancer. Methods The expressions of gastrokine 1 in gastric cancer and paracancerous tissues of 52 patients with gastriccancer were detected by real-time fluorescence quantitative polymerase chain reaction （RT-PCR） and immunohistochemistry. Meanwhile the relationship of the expression level of gastrokine 1 with clinicopathologic characteristics were analyzed. Results The expression levels of gastrokine 1 gene and protein in the gastric cancer tissues were significantly lower than those in the paracancerous tissues （P＜0.01）. No significant relationship was found between expression of gastrokine 1 gene and clinicopathologic features including tumor location， depth of invasion， differentiation， lymph node metastasis， tumor stage， gender， age， and preoperative peripheral blood CEA and CA19-9 levels （P＞0.05，respectively）. What’s more， the expression level of gastrokine 1 gene in gastric cancer tissues of Helicobacter pylori （HP）-positive patients was lower than that in the negative ones （P＜0.05）. Conclusions Gastrokine 1 may play a significant role as an anti-oncogene in the process of the formation and development of gastric cancer. Its effect may become weak due to HP infection in gastric cancer patients.