AIM: To investigate the role of SMADs singal pathway in rat myocardial hypertrophy. METHODS: The rat model of myocardial hypertrophy was produced by constriction of the abdominal aorta. The wet left vertricular/body weight ratio (LVMI) was measured. The expression of TGF-beta l and Smad 2, 3, 7 mRNA were assessed by RT-PCR. RESULTS: The LVMI and the expression of TGF-beta l and Smad 2, 3, 7 mRNA in cardiomyothy were increased in 3 day after the operation and continued at last 4 weeks. The peak expression of TGF-beta l and Smad 2, 3, 7 mRNA was in 2 weeks after operation. The expression of Smad 7 was increased in 3 days after operation, but the peak was in 1 week after operation, then decreased. CONCLUSION: The signal protein Smad 2, 3, 7 are involved in the progress of rat myocardial hypertrophy produced by constriction of abdominal aorta. [

OBJECTIVE:To study the comprehension of drug instructions in licensed physicians in order to promote the rational drug use.METHODS:The licensed physicians were investigated in a randomized sampling questionnaire.RESULTS:All the investigated doctors were found to lack comprehension of drug instructions,the overall rate of which was 11.33%.The rate of not understanding was significantly different between the physicians and the physicians higher than attending physicians in rank(P

Additive manufacturing (AM) is a collection of technologies based on the layer-by-layer manufacturing. Characterized by its direct manufacturing and rapidity, it has been regarded by the Economist Journal as one of the key techniques which will trigger the third industry reformation. The present article, beginning with a brief introduction of the history of AM and the process of its major technologies, focuses on the advantages and disadvantages and medical applications of the technique.
Medicine ; Printing, Three-Dimensional

Baculovirus gene expression is the most popular method to make target protein in cultured insect cells. To fast determine the generation of recombinant virus in cultured cells, donor plasmid of pFastBacI was modified by introducing egfp cassette. In the modified vector, egfp cassette was under the control of ie1 promoter, and target gene cassette was under the control of polyhedron promoter. To evaluate the convenience of the genetically modified donor plasmid used in eukaryotic expression, ns1 gene from Bombyx mori bidensovirus was ligated into the donor plasmid to generate recombinant plasmid pFastBacI-[P(ie1)-egfp-sv40]-[P(polh)-ns1-sv40]. Then the plasmid was transformed into DH10B competent cells containing Bm-Bacmid vector to produce the final recombinant Bm-Bacmid with the help of transposase. The resulting recombinant Bm-Bacmid was transfected into BmN cells to generate recombinant virus, which was easily and rapidly judged by green fluorescent signal observed in BmN cells. After infection for 96 h, the BmN cells were harvested and the total protein extracted from the infected BmN cells was subjected to Western blotting analysis. The result showed that a specific protein band about 36 kDa was detected, indicating that NS1 protein was successfully expressed in the BmN cells. In conclusion, the expression of NS1 protein with the modified expression system is useful for further research on the function of NS1 protein.
Animals ; Baculoviridae ; genetics ; Bombyx ; virology ; Cell Line ; Cells, Cultured ; Gene Expression ; Genetic Vectors ; Plasmids ; Promoter Regions, Genetic ; Transfection ; Viral Nonstructural Proteins ; biosynthesis ; genetics

Objective:To survey the expression of MMP-1,MMP-2 of human periodontal ligament cells(HPDLCs)treated by tea polyphenols(TP)and lipopolysaccharide(LPS).Methods:HPDLCs were in vitro cultured in vitro and treated by TP(200 μg/ml) and /or LPS(100 μg/ml)for 24,48 and 72 h respectively,the secretion of MMP-1 and MMP-2 were examined by ELISA,MMP-1 and MMP-2 mRNA expression was examined by real-time PCR.Results:The secression and mRNA expression of MMP-1 and MMP-2 of HPDLCs increased by LPS treatment and significantly inhibited by TP at the different times.Conclusion:TP can inhibit the col-lagen degradation of HPDLCs mediated by LPS.

To improve the expression of heterologous genes using baculovirus expression system, we constructed a novel shuttle vector based on the Bm-Bacmid. In the Bm-Bacmid, partial sequences of Chitinase and Cystein Protease were replaced with a tandem cassette of Cm and egfp through homologous recombination. Bombyx mori bidensovirus (BmBDV) ns1 under the control of polyhedrin promoter was inserted into the modified Bm-bacmid by transposition. For comparison, BmBDV ns1 under the control of polyhedrin promoter was also cloned in the wild type Bm-bacmid. The resulting Bm-bacmids were transfected into the cultured BmN cells to prepare recombinant virus to infect silkworms for expression of BmBDV ns1. Total proteins of hemocyte from infected silkworms were subjected to Western blotting and ELISA analysis. The yield of BmBDV NS1 1 with the modified vector was three times as much as that with the unmodified vector. The method to improve the yield of BmBDV NS1 in silkworms will facilitate the function and three-dimensional structure study of BmBDV NS1.
Animals ; Baculoviridae ; Bombyx ; virology ; Cell Line ; Chitinases ; Cysteine Proteases ; Genetic Vectors ; Insect Viruses ; Promoter Regions, Genetic ; Transfection

Bombyx mori bidensovirus (BmBDV) has been identified as causing chronic densonucleosis in Bombyx mori specifically. The replication mechanism of BmBDV remains unknown. Its genome comprises two single stands DNA (VD1 and VD2). In order to rescue infectious virions in vitro, we obtained the total viral DNA extracted from the BmBDV-infected larvae midguts, subsequently cloned the full-length sequence of BmBDV genome fragments by PCR and constructed recombinant plasmids pMD18T-VD1 and pUC-VD2. The linear genome fragments were obtained by digesting recombinant plasmids with corresponding restriction enzymes, and then collectively transfected BmN cells by the method of liposome-embedding. We determined the replication of the virus gene by PCR with the template of demethylated total DNA extracted from the post-transfect BmN cells. Meanwhile, we collected the total proteins from the post-transfect BmN cells and the larvae midgut of feeding the post-transfect BmN cells to perform Western blotting analysis, and detected the expression of viral genes. Here we firstly confirm that infectious virions can be rescued in BmN cells by linear co-transfect method.
Animals ; Bombyx ; DNA, Viral ; Densovirus ; growth & development ; Larva ; Transfection ; Virion ; Virus Cultivation

Objective:To study the effects of Ophiocordyceps xuefengensis on proliferation of DC -CIK cells and the activity of killing HepG-2 cells by DC-CIK cells.Methods:Peripheral blood mononuclear cells were routinely isolated and induced into DC and CIK.DC and CIK co-cultured by 1∶5 for 7 days,then Ophiocordyceps xuefengensis were added into medicine group ,observed the mor-phology of the cells on the tenth day and counted the DC-CIK number of each group.DC-CIK cells acted as effector cells and the HepG-2 cells as target cells , cck-8 method for the detection of DC-CIK in the killing rate of HepG-2.Results: The Ophiocordyceps xuefengensis was able to proliferate the DC-CIK dramatically ,the optimal concentration was 0.1 mg/ml.Cultivation of Ophiocordyceps xuefengensis induced DC-CIK cells on HepG-2 cells killing effect was better than that of the routine method of DC-CIK cells; the effection of Ophiocordyceps xuefengensis killing HepG-2 cells was not obviously.Conclusion: Ophiocordyceps xuefengensis can enhance the anti-tumor activity of DC-CIK mainly by promoting the proliferation of it.

Objective To establish a comprehensive evaluation indexing system to appraise the implications of prevention and treatment of HIV/AIDS ,and to calculate the weight of each indicator .Methods Based on the idea of performance and input-out-put ,professional consultation ,and Delphi method was determined as the evaluation index system ,analytical hierarchy process (AHP) was used to calculate the weight value for each indicator .Results The evaluation indexing system had been established af-ter three rounds of professional consultation .It contained two 1st class indicators ,six 2nd class indicators and thirty-one in 3rd class indicators .The weight value of each indicator was calculated .Conclusion The evaluation indexing system that has been established and the weight value quantities are of completeness ,practicality ,operability and logic .They have important value for application in the future .

Objective To investigate the safety and efficacy of decitabine combined with CAG regimen in treatment of acute myeloid leukemia (AML) ineligible for conventional chemotherapy. Methods The data of 20 cases with AML ineligible for conventional chemotherapy from January 2013 to May 2015 were retrospectively analyzed. Decitabine combined with CAG regimen was used during induction therapy. The primary induction regimen was used 26 times after remission, the standard 3+7 regimen were used 7 times, and intermediate-dose cytarabine were used 3 times. The total course of treatment included 2-8 cycles. Results All of the 20 patients completed the first cycle of induction therapy, including 11 cases of complete remission (CR), 5 cases of partial remission and no response in 4 cases, and the overall response rate (ORR) was 80 % (16/20). ORR was 69.2 % (9/13) and 100.0 % (7/7) in high-risk group and middle-low risk group respectively. ORR was 60.0%(6/10) in AML evolving from MDS. 8 patients were infected during the induction therapy and the infection rate was 40.0% (8/20). 2 patients were died of pulmonary infection. The median number of suspended red blood cell and platelet infused were (9.1±5.7) U and (57.5±51.9) U respectively. Neutrophil recovery time was (8.7±5.6) days during induction therapy. All patients were followed up for at least 1 year, and 12 cases were dead. Overall survival rate was 85.0%at 3 months, 80.0%at 6 months, and 40.0%at 1 year. While in 12 CR patients relapse-free survival rate was 75.0%at 3 months, 75.0%at 6 months,and 65.6%at 1 year respectively. Conclusion Decitabine combined with CAG regimen with high remission rate and well tolerance, can be used as a first therapy for AML ineligible for conventional chemotherapy.