Effects of Shenggu Injection (生骨注射液) on mRNA Expression of Vascular Endothelia Growth Factor in Rat Osteoblasts in vitro

Objective： To study the effects of Shenggu injection （生骨注射液,SGI） on mRNA expression of vascular endothelia growth factor （VEGF） in rat osteoblasts in vitro and to explore its possible molecular mechanisms in promoting fracture healing. Methods： Rat osteoblasts cultured in vitro were stimulated with SGI according to the protocol. The expression levels of VEGF mRNA in the cells in every group were examined by reverse-transcriptase ploymerase chain reaction （RT-POR）. Results： When osteoblasts were stimulated with different concentrations of SGI for 5 days, the expression of VEGF mRNA peaked with 1 mg/ml SGI on the 5th day. When treated with 1 mg/ml SGI from the 1st to the 5th day, the expression of VEGF mRNA increased gradually with the increase of culturing time. Conclusion： SGI could promote significantly the expression of VEGF mRNA in rat osteoblasts in vitro. The levels of expression of VEGF mRNA changed along with different concentrations and stimulating time of SGI.

Cloning and Identification of A Novel Variant of Human Vascular Endothelial Growth Factor

A novel variant of human vascular endothelial growth factor (h'VEGF165) cDNA was amplified by nested PCR method from the HL60 1 cells and was cloned into a eukaryotic expressing vector pcDNA 3 to construct a recombinant plasmid pCD-h'VEGF165. The amplified h'VEGF165 cDNA fragment was identified by enzyme digestion and DNA sequencing methods. Also, wild-type hVEGF165 cDNA was obtained, identified and cloned into a eukaryotic expressing vector pcDNA3 by using the same methods. The results of DNA sequencing showed that h'VEGF165 cDNA cloned from HL60 1 was 600 bp in size with 8 % of the base sequence in h'VEGF165 cDNA being changed as compared with the base sequence in the wild-type hVEGF165 cDNA. The results of sequencing of hVEGF165 which was cloned from HL60 by us were consistent with the reports completely.

Effects of recombinant adeno-associated viruses expressing human vascular endothelial growth factor 165 on spinal cord injury

Numerous studies have confirmed that vascular endothelial growth factor （VEGF） improves the function of neural cells following spinal cord injury （SCI）. However, some studies have also verified that VEGF cannot significantly induce the increase in vascular density at or surrounding the lesion, and that VEGF therapy exacerbated secondary damage following SCI. Based on the dual effects of VEGF on SCI, we constructed the recombinant adeno-associated viruses （rAAV）-hVEGF165-IRES-human recombinant green fluorescent protein （hrGFP） （AAV-VEGF） and rAAV-IRES-hrGFP （AAV-GFP）. Our results suggested that rAAV expressed hVEGFles, and a low dose of VEGF relieved increased vascular permeability, improved microcirculation in the local spinal cord, lessened spinal cord edema, and decreased neuronal apoptosis. These results verified that the releasing effects of the rAAV virus vector had protective effects on the spinal cord.

Effects of Danshen Injection on the Malignant Obstructive Jaundice in the SD Rat Model

To observe the effects of Danshen on the growth of hepatocellular carcinoma in the SD rats, a model of malignant obstructive jaundice was established by inoculation of transplanted tumor into the hepatic portal with the walker-256 hepatocarcine line, which resulted in the obstruction by the infiltration and metastasis of hepatocellular carcinoma. SD rats were divided into 4 groups： the rats were treated by 0.9 % NS （n=24, control group）, inosine＋vitamin C （n=40, InV group）, Danshen （n=40, DS group） and 5-FU （n=40, 5-FU group）, respectively. The liver function, morphological changes and the expressions of PCNA, VEGF and ICAM-1 in carcinoma foci, peri-carcinoma tissues, adjacent lobe （left-internal lobe） and lung tissues were observed after the treatment with the 4 agents. Our results showed that the protective effect of Danshen on liver function was significantly better than that of NS and 5-FU （P〈0.01）. No significant difference in protective effect was observed between DS group and lnV group （P〉0.05）. Danshen also provided protective effect on the morphological damage of liver caused by obstructive jaundice. The rates of carcinoma-inhibition and metastasis inhibition were significantly higher than those of NS and inosine＋vitamin C （P〈0.01）. No significant difference in this regard existed between DS group and 5-FU group （P〉0.05）. The expressions of PCNA,VEGF and ICAM-1 PCNA, VEGF and ICAM-1 in carcinoma foci, peri-carcinoma tissues, adjacent lobe （left-internal lobe） and lung tissues were lower than those in control group and InV group, with the differences being significant （P〈0.01）. No significant differences were found between DS group and 5-FU group in the expression levels of PCNA and VEGF （P〉0.05） but ICAM-1 （P〈0.05）. It is concluded that Danshen injection not only has protective effects on liver injury caused by obstructive jaundice, but can inhibit the proliferation and growth of hepatocarcinoma, interfere with the vascularization of tumors, prevent recurrence and metastasis of hepatocarcineoma.