Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state. Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices.
Adenoviridae ; genetics ; Animals ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Osteoclasts ; metabolism ; Osteoprotegerin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the cytotoxic activity against tumor cells and cytokines production of spleen cells induced in vitro by murine 4-1BBL gene transfected Hepa1-6.

METHODSThe eukaryotic expression vector pCDNA3.1(+)-m4-1BBL was transfected into murine hepatocellular carcinoma cell line Hepa1-6 by Liposomes. Then the transfected cells were selected in medium containing G418 (400 - 800 micro g/ml) and termed as Hepa1-6-m4-1BBL. The TCV-m4-1BBL was obtained by treating them with mitomycin (MMC). Cocultivation TCV with syngeneic murine spleen cells, then the lymphocytes were tested for cytotoxic activity against Hepa1-6-wt cells and the supernatants were harvested for detecting the cytokines (IL-2, TNF-alpha and GM-CSF).

RESULTSHepa1-6-m4-1BBL cells expressed 4-1BBL protein with highest cell surface level. The 4-1BBL mRNA could still be detected in the cells when cultured 48 h after treated with MMC (80 mg/L). Comparing with TCV-Hepa1-6, the tumor cell vaccine derived from Hepa1-6-m4-1BBL (TCV-m4-1BBL) could induce a more efficient cytotoxic activity of syngeneic murine lymphocyte against its parental tumor cell Hepa1-6 (P < 0.05), but not against non-parental tumor cell H22 and NIH3T3. Higher levels of IL-2, TNF-alpha and GM-CSF were released by the splencytes after stimulated by TCV-m4-1BBL.

CONCLUSIONSThese results suggest the expression of m4-1BBL by tumor cells is effective in inducing antitumor immune responses.

4-1BB Ligand ; Animals ; Cancer Vaccines ; immunology ; Female ; In Vitro Techniques ; Liver Neoplasms, Experimental ; immunology ; Mice ; Mice, Inbred C57BL ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Tumor Necrosis Factors ; genetics ; physiology

This paper aims to summarize the achievements during the implementation process of good agricultural practice (GAP) in Chinese Materia Medica (CMM), and on basis of analyzing the existing problems of GAP, to propose further implementation of GAP in TCM growing. Since the launch of GAP in CMM growing ten years ago, it has acquired great achievements, including: (1) The promulgation of a series of measures for the administration of the GAP approval in the CMM growing; (2) The expanded planting area of CMM; (3) The increased awareness of standardized CMM growing among farmers and enterprises; (4) The establishment of GAP implementation bases for CMM growing; (5) The improvement of theory and methodology for CMM growing; (6) The development of a large group of experts and scholars in GAP approval for CMM production. The problems existing in the production include: (1) A deep understanding of GAP and its certification is still needed; (2) The distribution of the certification base is not reasonable; (3) The geo-economics effect and the backward farming practices are thought to be the bottlenecks in the standardization of CMM growing and the scale production of CMM; (4) Low comparative effectiveness limits the development of the GAP; (5) The base of breeding improved variety is blank; (6) The immature of the cultivation technique lead to the risk of production process; (7) The degradation of soil microbial and the continuous cropping obstacle restrict the sustainable development of the GAP base. To further promote the health and orderly GAP in the CMM growing, the authors propose: (1) To change the mode of production; (2) To establish a sound standard system so as to ensure quality products for fair prices; (3) To fully consider the geo-economic culture and vigorously promote the definite cultivating of traditional Chinese medicinal materials; (4) To strengthen the transformation and generalization of basic researches and achievements, in order to provide technical support for the CMM production; (5) To deepen the understanding of GAP, to vigorously promote ecological planting and precision agriculture, in order to overcome the continuous cropping obstacle. The authors think that despite the fact that we are still facing with a huge array of management and technological problems, the GAP in the CMM growing has already enjoyed widespread support and showed great potential. In the future, with people's deeper understanding of GAP and the great progress of the science and technology, the GAP will constantly be fused with the theory, methodology and technology in the modern agriculture like precision agriculture, eco-agriculture and etc.
Agriculture ; economics ; methods ; standards ; trends ; China ; Drugs, Chinese Herbal ; analysis ; economics ; standards ; Humans ; Materia Medica ; chemistry ; economics ; standards ; Plants, Medicinal ; chemistry ; growth & development

OBJECTIVETo investigate the association between the polymorphism of the thromboxane synthase gene and Uigur patients with myocardial infarction (MI) in Xinjiang.

METHODSThree hundred and fifteen patients with MI and 218 healthy control subjects were detected by polymerase chain reaction and restriction fragment length polymorphism. The serum thromboxane B2 (TXB2) in all subjects was detected with radioimmunoassay kit.

RESULTSThe genotype distributions of the MI group and control group were in Hardy-Weinberg equilibrium (Chi-square=0.375,0.029, P>0.05). The frequencies of CC and TC were 0.933 and 0.067 in MI group while they were 0.977 and 0.023 in controls. There was significant difference in frequencies of the TC genotype and T allele but no difference in frequencies of CC genotype between controls and MI cases. There was significant difference in serum TXB2 level between the MI and control group (P<0.05), and between individuals of the TC and CC genotypes (P<0.05). The serum TXB2 level in the MI cases with TC genotype was increased compared with that of other genotypes (P<0.05).

CONCLUSIONThe TC genotype and T allele of thromboxane synthase gene might be risk factors of MI in Uigur population in Xinjiang, which might result from the increased serum TXB2 level.

Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; Base Sequence ; Case-Control Studies ; China ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Myocardial Infarction ; blood ; enzymology ; ethnology ; genetics ; Polymorphism, Genetic ; Thromboxane B2 ; blood ; Thromboxane-A Synthase ; genetics

OBJECTIVETo investigate the association between the polymorphism of thromboxane synthase gene (CYP5A1) and myocardial infarction (MI) of Uigur nationality patients in Xinjiang.

METHODSRs10487667 site polymorphism in CYP5A1 gene of 318 patients with MI (MI group) and 232 healthy control subjects (control group) were analyzed by polymerase chain reaction and restriction fragment length polymorphism. The serum thromboxane B(2)(TXB(2)) concentration was also detected in all subjects. The relationship of multiple factors and myocardial infarction was evaluated comprehensively by non-condition logistic regression analysis.

RESULTSThe frequencies of CYP5A1 gene Rs10487667 site polymorphism in MI group and control group were: GG type 0.204 (65/318) and 0.155 (36/232), GT type 0.553 (176/318) and 0.466 (106/232), TT type 0.242 (77/318) and 0.379 (88/232), respectively. There was significant difference in frequencies of GG genotype (χ(2) = 12.193, P = 0.002) between two groups and G allele frequency in MI group (0.481 (306/636)) was significant higher than control group (0.388 (180/464)) (χ(2) = 9.449, P = 0.021), but no difference in frequencies of GT and TT genotypes (χ(2) = 0.699, P > 0.05)between controls and MI cases. There was significant difference in serum TXB(2) level between MI ((184.3 ± 34.7) pg/ml) and control ((124.3 ± 28.1) pg/ml) groups (t = 5.503, P = 0.034). In the case and control group, the serum TXB(2) level of the person with GT + GG genotype ((164.21 ± 22.56) and (134.26 ± 19.83) pg/ml)) was significant higher than those of TT genotypes ((113.67 ± 54.23) and (98.54 ± 13.11) pg/ml) (t values were 5.433 and 5.108, respectively, both P values < 0.05). Logistic regression analysis showed that the T allele of the CYP5A1 gene was one independent risk factor of MI (OR = 1.673, 95%CI: 1.020 - 2.156) after adjustment of risk factors.

CONCLUSIONRs10487667 polymorphism in CYP5A1 gene might be a risk factor of MI in Uigur population in Xinjiang, which maybe related with the significant high serum TXB(2) level.

Alleles ; Case-Control Studies ; China ; epidemiology ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; epidemiology ; genetics ; Polymorphism, Genetic ; Thromboxane B2 ; blood ; Thromboxane-A Synthase ; genetics

OBJECTIVEIn order to explore the existence of SARS coronavirus (Co-V) and/or its RNA in sewage of hospitals administered SARS patients.

METHODSA novel electropositive filter was used to concentrate the SARS-CoV from the sewage of two hospitals administered SARS patients in Beijing, including twelve 2,500 ml sewage samples from the hospitals before disinfection, and ten 25,000 ml samples after disinfection; as well as cell culture, RT-PCR and sequencing of gene to detect and identify the viruses from sewage.

RESULTSThere was no live SARS-CoV detected in the sewage in this study. The nucleic acid of SARS-CoV had been found in the 12 sewage samples before disinfection from both hospitals by semi-nested PCR. After disinfection, SARS-CoV RNA could only be detected from the samples from the 309th Hospital, and the others were negative.

CONCLUSIONIt provides evidence that there is no live SARS-Cov in the sewage from hospitals with SARS patients though SARS-CoV RNA can be detected.

Hospitals ; Humans ; Nucleocapsid ; analysis ; RNA, Viral ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; virology ; Sewage ; virology

OBJECTIVE: To investigate the effect of galectin-1 on biological behaviors of lung adenocarcinoma cells and the underlying mechanism. METHODS: The expression levels of galectin-1 mRNA were detected in 8 pairs of lung adenocarcinoma tissues and adjacent normal tissues and in lung adenocarcinoma cell lines A549 and H1299 and normal bronchial epithelial cell line BEAS-2b using qRT-PCR. The effect of galectin-1 knockdown by RNA interference on the proliferation, invasion and migration abilities and apoptosis of lung adenocarcinoma cells were examined using CCK8 assay, Transwell assay, scratch assay and flow cytometry. Western blotting was performed to detect the expressions of apoptosis-related proteins BAX, BCL-2, and caspase-3 and the proteins involved in the AKT and ERK pathways. RESULTS: The mRNA expression of galectin-1 was significantly increased in lung cancer tissues and lung adenocarcinoma cell lines (P < 0.05). In lung adenocarcinoma cells, galectin-1 knockdown significantly inhibited cell proliferation, migration and invasion and obviously increased cell apoptosis rate (P < 0.05), causing also significant reduction of the phosphorylation level of ERK signaling pathway (P < 0.05). CONCLUSION Galectin-1 knockdown inhibits proliferation, migration, and invasion and promotes apoptosis of lung adenocarcinoma cells, and this effect is mediated probably by inhibition of the phosphorylation levels of the ERK pathway.
Humans ; Adenocarcinoma of Lung ; Apoptosis ; Lung Neoplasms ; MAP Kinase Signaling System ; Cell Proliferation

ObjectiveTo clarify the medication regularity of WU Zhao-dong，a famous chief physician in traditional Chinese medicine （TCM） of Jiangxi province， and investigate the potential mechanism of potential new prescriptions against chronic renal failure （CRF）. MethodThe outpatient prescriptions of WU Zhao-dong from July 2019 to July 2021 were collected. Data mining was carried out by using the Traditional Chinese Medicine Inheritance Auxiliary Platform （V 2.5） to analyze the medication frequency and drug association and obtain potential new prescriptions. The interaction between drug targets in new prescriptions was analyzed by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP），STRING，and Kyoto Encyclopedia of Genes and Genomes（KEGG）， followed by verification by molecular docking and experiments. ResultA total of 200 prescriptions were screened out， with 217 Chinese medicinal drugs involved， and eight new potential prescriptions were derived. To be specific， Prescription 1： Armeniacae Semen Amarum-Astragali Radix-Platycodonis Radix-Angelicae Sinensis Radix-Smilacis Glabrae Rhizoma-fried Atractylodis Macrocephalae Rhizoma， Prescription 2： Saposhnikoviae Radix-Schizonepetae Herba-Kochiae Fructus-Asteris Radix et Rhizoma-Menthae Haplocalycis Herba，Prescription 3：Armeniacae Semen Amarum-Asteris Radix et Rhizoma-Platycodonis Radix-Eriobotryae Folium-prepared Ephedrae Herba， Prescription 4：Perillae Caulis-Codonopsis Radix-Coptidis Rhizoma-Pseudostellariae Radix， Prescription 5：Ecliptae Herba-Astragali Radix Praeparata Cum Melle-Dryopteridis Crassirhizomatis Rhizoma-Rosae Laevigatae Fructus-Coicis Semen-Ligustri Lucidi Fructus， Prescription 6： Lycopi Herba-Lonice Raejaponicae Caulis-Trachelospermi Caulis et Folium-Alismatis Rhizoma， Prescription 7：Scutellariae Radix-Hirudo-Paeoniae Radix Rubra-Eriobotryae Folium-Glehniae Radix， Prescription 8：Glycyrrhizae Radix et Rhizoma-Scrophulariae Radix-Chrysanthemi Indici Flos-Smilacis Glabrae Rhizoma- Serissae Herba. In Prescription 1，18 main chemical components were screened out. Eighty targets of active components of Prescription 1 were predicted， and 37 potential targets for the treatment of CRF were obtained， including interleukin （IL）-6， vascular endothelial growth factor （VEGF）， cysteinyl aspartate-specific protease-3 （Caspase-3）， nitric oxide synthase 3 （NOS3）， and hypoxia-inducible factor-1 （HIF-1）. The KEGG pathways involved in the targets of Chinese medicinal drugs and disease mainly included the signaling pathways of lipid and atherosclerosis，NF-κB， Toll-like receptors， and HIF-1. Prescription 1 significantly decreased serum creatinine and urea nitrogen， and increased the content of NO and NOS3 in renal tissues of CRF rats. ConclusionPrescription 1 shows the multi-component and multi-target characteristics of action，and its mechanism may be related to its inhibition of renal fibrosis，anti-inflammation，improvement of intestinal microecology，and improvement of renal hypoxia and ischemia.

OBJECTIVESTo evaluate the changes of the position of medulla oblongata and cerebellum following posterior fossa decompression (PFD), and to investigate their influences on the prognosis of the syringomyelia in adolescents with Chiari malformation (CM).

METHODSA retrospective review was performed on all CM patients between September 2006 and September 2011. A subset of 46 patients, including 25 male and 21 female patients, was finally enrolled according to the inclusion criteria. The initial age and duration of follow-up averaged 13.9 years (range, 10-17 years) and 13 months (range, 6-52 months), respectively. On mid-sagittal MRI, the following parameters were evaluated pre- and postoperatively (follow-up ≥ 6 months): the longitudinal and transverse position of bulbopontine sulcus, the fourth ventricle vertex, the lower extreme of cerebella tonsil, the cervico-medullary angle, the maximal syrinx/cord(S/C) ratio and the syrinx length. Changes in these parameters were analysed using the paired samples t test, and for these reaching statistical significances, an additional bivariate correlation analysis was performed to investigate their relation with syrinx resolution.

RESULTSAt the latest follow-up, upward shifting of the bulbopontine sulcus was observed in 31 patients(67.4%), with upward shifting of the lower extreme of cerebella tonsil presenting in 35 patients(76.0%). The maximal S/C ratio and the syrinx length were significantly improved postoperatively (t = 7.114 and 7.816, P = 0.000).Significant resolution of the syrinx was demonstrated in 40 patients(86.9%), and more specifically, the average improvement rates of the maximal S/C ratio and the syrinx length were 32% ± 30%and 43% ± 33%, respectively. In addition, the bivariate correlation analysis revealed that syrinx resolution was significantly correlated with the upward shifting of the bulbopontine sulcus (r = 0.332, P = 0.027) and lower extreme of cerebella (r = 0.298, P = 0.044) .

CONCLUSIONThe upward shifting of the bulbopontine sulcus and the lower extreme of cerebella tonsil might be implicated in the mechanisms of postoperative syrinx resolution.

Adolescent ; Arnold-Chiari Malformation ; complications ; surgery ; Child ; Decompression, Surgical ; Female ; Humans ; Male ; Medulla Oblongata ; pathology ; Postoperative Period ; Retrospective Studies ; Syringomyelia ; complications ; surgery ; Treatment Outcome