[ ABSTRACT] AIM:To investigate the therapeutic effects of lentivirus-mediated shRNA targeting growth hormone secretagogue receptor 1a (GHSR1a) on colorectal cancer cell line SW480 both in vitro and in vivo.METHODS: Human GHSR1a sequence was used for the design of shRNA targeting GHSR1a, which was introduced to lentivirus, followed by transfection into SW480 cells.CCK-8 assay was performed to detect cell viability.The mRNA expression of GHSR1a and PTEN in colorectal cancer cells was detected by RT-PCR.The protein levels of GHSR1a, ghrelin, PTEN, p-AKT and p53 were determined by Western blot.Stable GHSR1a silencing in SW480 xenografts in nude mice was established.After the mice were sacrificed and weighted, immunohistochemistry was used to detect the positive expression of Ki-67 and PTEN in the tumors.RESULTS:GHSR1a was over-expressed in the malignant cells in comparison with the normal cells in vitro. The tumor specific lentivirus-mediated shRNA targeting GHSR1a gene and GHSR1a knockdown SW480 cells were success-fully constructed.After transfection with GHSR1a shRNA, the expression of GHSR1a at mRNA and protein levels was markedly inhibited in the SW480 cells.Furthermore, GHSR1a silencing by specific shRNA showed increased PTEN, inhi-bition of AKT phosphorylation and promotion of p53 release in the SW480 cells.In vivo results demonstrated that down-re-gulation of GHSR1a in the SW480 cells significantly decreased the expression of Ki-67 and increased the expression of PTEN in the tumor tissues, leading to a marked reduction in tumor weight in comparison to blank control or negative control
tumors.CONCLUSION:Down-regulation of GHSR1a by shRNA technique inhibits the growth of colorectal cancer cell line and xenograft tumor through activation of the PTEN/PI3K/AKT signaling pathways.

Understanding inter-species transmission of influenza viruses is an important research topic. Scientists try to identify and evaluate the functional factors determining the host range of influenza viruses by generating the recombinant viruses through reverse genetics in laboratories, which reveals the viruses' molecular mechanisms of infection and transmission in different species. Therefore, the reverse genetic method is a very important tool for further understanding the biology of influenza viruses and will provide the insight for the prevention and treatment of infections and transmission. However, these recombinant influenza viruses generated in laboratories will become the potential threat to the public health and the environment. In this paper, we discussed the biological safety issues of recombinant influenza viruses and suggested we should set up protocols for risk management on research activities related to recombinant highly pathogenic influenza viruses.
Influenza A Virus, H5N1 Subtype ; genetics ; Laboratories ; Microbiology ; Orthomyxoviridae ; genetics ; Public Health ; Reassortant Viruses ; genetics ; Recombination, Genetic ; Safety

Objective To explore the method and superiority of laparoscopic management of children's varicocele. Methods High position ligation of varicocele under laparoscope was performed and internal spermatic arteries were reserved intraoperatively. Results 32 cases had smooth recovery after operation, and no complication occurred. The hospital stay was (1～3)days postoperatively. 29 cases had been followed up for 6 months～1 year and no relapse and testis atrophy occurred. Conclusions The method has the advantages of simpleness, minimal invasion and quicker recovery. Reservation of internal spermatic artery can prevent testis atrophy.

OBJECTIVE:To observe the effect of Shuanghuang tongfeng capsule(SHTF) on the gouty arthritis.METH-ODS:The gouty arthritis model of rats and rabbits were induced using sodium urate(MSU). The swollen degree of the rat foot,and the amount of synovial fluid,the leucocyte count in synovial fluid and the degree of inflammation of the synovial tissue of joints in rabbits were measured.The effect of SHTF on gouty arthritis was observed. RESULTS:The swollen degree of the foot of rat was lessened obviously by SHTF capsule(5、10、20 g?kg-1,ig).The amount of synovial fluid and the leucocyte count in synovial fluid of the rabbit were decreased significantly by SHTF capsule (4、8、16 g?kg-1,po).The inflammation of the synovial tissue in rabbits was improved obviously by SHTF capsule (8 and 16 g?kg-1,po). CONCLUSION: SHTF capsule have remarkable protective effect on the gouty arthritis induced by sodium urate.

Objective To investigate whether the effect of contrast media on renal tubular cell apoptosis is mediated via mitogen activated protein kinase(MAPK) activation. Methods NRK 52E cells were incubated in cell culture media[+1%fatel calf serum (FCS)] containing buffer (control) or increasing concentrations of diatrizoate or iohexol for various time periods. In separate experiments, NRK 52E cells were incubated with diatrizoate in the presence of 10 or 20 ?mol/L SB203580, an inhibitor of p38 MAPK, for 4 h. Cell membrane integrity was assessed by trypan blue exclusion experiment. Apoptosis was assessed by flow cytometric DNA analysis, FITC Annexin Ⅴbinding/PI staining and electron microscopy. MAPK activity and phosphorylation was evaluated by immunoprecipitation and Western blot analysis. Results Diatrizoate, a kind of hyperosmolal contrast media, induced apoptosis in cultured NRK 52E cells in an osmotic pressure and duration dependent manner. Iohexol, another kind of less hyperosmolal contrast media, could not induce NRK 52E cells apoptosis. In NRK 52E cells incubated with diatrizoate, the level of p38 MAPK phosphorylation and activity increased after 15 min, and remained elevated even at 1 h. In contrast, the level of total p38 protein was not changed in whole experimental period. SB203580 significantly inhibited apoptosis in NRK 52E cells incubated with diatrizoate. The level of p44/p42 MAPK phosphorylation was not changed in whole experimental period. Conclusions Contrast media induces apoptosis in cultured NRK 52E cells in an osmotic pressure and duration dependent manner, which is related to the hypertonicity of contrast media. Contrast media induces apoptosis in NRK 52E cells via activation of p38 MAPK.

Objective:To observe the effects of Shen Hong Bu Xue Granule(SHBXG) on the expression levels of erythropoietin(EPO)mRNA in kidney tissue and granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA in bone marrow tissue of the mice with blood deficiency,and to investigate the protective effect of SHBXG on the blood deficiency mice and its mechanism.Methods:The mouse model of blood deficiency was established with acetylphenylhydrazine(APH) and cytoxan(CTX).A total of 60 mice were divided into blank control group,model group,Compound E-jiao Slurry group and low,middle,high doses of SHBXG groups(n=10).The serum,kidney and bone marrow tissues from all the mice were collected after 14 d consecutive administration.The levels of red blood cells(RBC),hemoglobin level(HGB),hematocrit(HCT),leukocytes(WBC),and platelet(PLT) in blood of the mice were detected by automatic blood analyzer;the levels of serum EPO and GM-CSF of the mice in various groups mice were detected by ELISA method;the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice were detected by RT-PCR method.Results:The results of automatic blood analyzer showed that the levels of RBC,HGB,HCT,and PLT of the mice in Compound E-jiao Slurry group and different doses of SHBXG groups were increased significantly compared with model group (P<0.05 or P<0.01);the levels of WBC of mice in Compound E-jiao Slurry group and high dose of SHBXG group were increased significantly compared with model group(P<0.05).The ELISA results showed that the levels of serum EPO and GM-CSF of the mice in Compound E-jiao Slurry group and different doses of SHBXG groups were increased significantly compared with model group(P<0.05).The PC-PCR results showed that the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice in Compound E-jiao Slurry group and middle and high doses of SHBXG groups were increased significantly compared with model group(P<0.05 or P<0.01).Conclusion:SHBXG could improve the blood deficiency symptom in the mice with blood deficiency,and its mechanism may be related to increasing the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice.

Based on the work characteristics of disease and control of People's Liberation Army (PLA), the paper discusses the establishment of quality control system at Institute of Disease and Control of PLA and the subsequent effects. This could provide some reference for improvement disease control of PLA.

Objective To study the type variation of microglial activation in spinal dorsal horn of rats after sciatic nerve injury.Methods Healthy adult male Sprague-Dawley rats were randomly divided into the control and experimental groups, 24 rats in each group.The experimental group underwent ligation of sciatic nerve trunk to generate nerve injury in the rats.The pain behavior in the rats was measured at the 1th, 7th and 14th postoperative days, and the changes of microglial activation in the rat lumbar spinal cord dorsal horn was detected by immunofluorescence staining.qRT-PCR assay was used to validate the activation trends of M1 and M2 types of microglia cells.Results No significant changes were found in the microglial cells in the spinal cord dorsal horn of rats in the sham-operation group during 14 days after operation.In the sciatic nerve ligation group at 1 day after operation, no significant change was observed in the number of microglial cells, but the expression of marker of M1 microglia was significantly increased.At 7 and 14 days after operation, the number of microglial cells and the expression of M1 microglia marker in the spinal cord dorsal horn were increased significantly.Conclusions Microglia activation in the spinal dorsal horn starts at the first day after sciatic nerve injury, and lasts at least for two weeks after the operation.M1 microglia activation dominates during this period.

Objective To discuss the surgical treatment for huge hepatoblastoma in children,and the technique of hepatectomy without blockade of the blood supply to the remained liver lobes.Methods We reviewed 12 cases of huge hepatoblastorna who had been operated from July 2001 to January 2009 in our hospital.The mean age of the children was 3.2 years(range,11 months to 12 years).The diameter of the tumor was from 10 to 23 cm.3～7 cycles of chemotherapy was routinely administrated before operation.When the tumor reduced to a certain size that radical resection could be performed safely,regular hepatectomy was conducted.Hepatoblastoma resection without blocking the blood supply to the remained liver lobes was performed in every patient.Results The operations were successfully accomplished in all the 12 children.5 cases received right trihepatectomy (segment Ⅳ,Ⅴ～Ⅷ),4 cases received right hemihepatectomy(segment Ⅴ～Ⅷ),and the other 3 cases received Ⅳ,Ⅶ,Ⅷ segmentectomy,right Ⅴ,Ⅵ segmentectomy,and left hemihepatectomy respectively.The intraoperative hemodynamic parameters were stable,and there was no perioperative mortality.Postoperative chemotherapy wag routinely administrated.The follow-up period varied from 2 to 92 months.11 children survived and were disease free,among those 6 children have survived for more than 5 years.One child had brain and lung metastasis 5 months post operation,and died 7 months post operation. Conclusion Preoperative chemotherapy administrated to children with huge hepatoblastoma can reduce the tumor size and render tumor reseetable.Hepatoblastoma resection without blocking the blood supply to the remained liver lobes is a safe and feasible surgical technique.

Objective To observe age-dependent feature of damage of hippocampus to different maturational stages rats after kindling repeated seizures.Methods The effects of 5 daily pentylenetetrazol-induced convulsions in different rats beginning at postnatal day 10,20,60(P10,P20,P60)were evaluated.In the 3 groups,Thionin staining method was utilized to observe morphological changes and cell counting of dentate granule cells,CA3,CA1,and hilar neurons.Timm's method of silver sulfide staining was adopted to observe the mossy fiber sprouting.Results 1.Cell counting of CA1,CA3 and hilar neurons in P10 and P20 groups demonstrated no differences from controls in rats,whereas P60 with daily seizures had a significant decrease in CA1,CA3 neurons(8.22?1.88,5.62?1.68 vs 6.31?1.50,3.62?1.40)(t=2.246,2.587 Pa