Drug Resistance, Neoplasm ; Humans ; Multiple Myeloma

Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

Human leukocyte antigen (HLA) is an important immunogenetic system. It plays a key role in immunoresponsiveness and hematopoietic stem cell and organ transplantation. The development of modern medicine and tissue typing promotes the combination of molecular biology techniques with HLA typing, and improves the accuracy and practicability of HLA typing. In this review, recent advances were concisely introduced in research of HLA typing on the application of restriction fragment length polymorphism (RFLP), sequence specific oligonucleotide probe (SSOP), single strand conformational polymorphism (SSCP), sequence specific primer (SSP), flow cytometry, gene chip, sequencing based typing, virtual DNA analysis and so on.
DNA ; analysis ; genetics ; Flow Cytometry ; methods ; HLA Antigens ; genetics ; Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational

In the present study, the antibacterial spectrum of turmeric extract was analyzed by measuring the minimum inhibitory concentration (MIC), and the antibacterial mechanism of turmeric extract was elaborated by determining its effects on the permeability and integrity of the cytoplasmic membrane, energy metabolism, and the morphology of the tested bacteria (Bacillus subtilis) with the highest susceptibility to the extract. The results showed that turmeric extract possessed broad-spectrum antibacterial activity against Gram-positive, Gram-negative bacteria and fungi, especially against Bacillus subtilis, with the MIC of 0.5 mg·mL^-1. Turmeric extract disrupted the liposome membrane and released calcein encapsulated within it. The permeability of the bacterial cell membrane was increased, leading to leakage of intracellular K⁺, Ca²⁺and cell wall proteins, and the integrity of the cell membrane was broken down, causing leakage of intracellular proteins and polysaccharides. Scanning electron microscope observation confirmed that the bacteria treated with turmeric extract was severely deformed. Simultaneously, cellular energy metabolism was affected, resulting in a significant reduction in the intracellular ATP content and ATPase activity in bacteria. In summary, the antibacterial mode of turmeric extract is closely related to its disruption of bacterial membrane structure.

Objective To investigate the expression of transforming growth factor β1,transforming growth factor beta receptor(TBR)Ⅰ,TβR Ⅱ,Smad4 and C-Jun in rats with nonalcoholic fatty liver disease(NAFLD)and to find out the mechanisms of liver fibrosis in patients with NAFLD.Methods A total of 18 male SD rats were randomly divided into normal control group(n=9)and model group(n=9).The rats in control group were fed with normal diet,and those in model group were fed with fat-rich diet(consisted of 10%lard oil+2%cholesterol).An rats were sacrificed at the 20th week.The levels of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were examined by RT-PCR.The expressions of TGFβ1 and Smad4 in liver tissue were detected by immunohistochemistry.The expression of C-Jun protein was detected by Western blotting.Results The NAFLD model was successfully established.The immunohistochemistry examination revealed that TGFβ1 and Smad4 were expressed weekly in control group,but strongly expressed in model group.RT-PCR showed that A values of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were 0.46±0.12,5.z4±2.70 and 3.35±1.95,respectively,in model group,which were higher than those in control group(0.21±0.09,1.36±0.77 and 0.52±0.19,all P values＜0.01).The Western blotting results demonstrated that the expression of C-Jun protein in model group(0.93±0.41)was higher than that in control group (0.32±0.25,P=0.001).Conclusion TGFβ1/Smad4 signal pathway might be involved in the development of hepatic fibrosis in NAFLD.Blocking TGFβ1/Smad4 signal pathway will be helpful in treatment of NAFLD.

Determination of postmortem interval (PMI) is one of the most valuable subjects in forensic practice. It, however, is often very difficult to accurately determine the PMI in daily practice. Forensic DNA technology has recently been used to estimate the PMI. It has certain advantage to traditional methods. This article reviews this technology with respect to its invention, development, advantage, disadvantage, and potential future applications with emphasis on correlation of DNA degradation and PMI.
Animals ; Bone Marrow Cells/metabolism* ; Cell Nucleus/metabolism* ; DNA/metabolism* ; Flow Cytometry ; Forensic Medicine/methods* ; Hepatocytes/metabolism* ; Humans ; Image Processing, Computer-Assisted/methods* ; Myocardium/metabolism* ; Postmortem Changes ; Spleen/metabolism* ; Time Factors

Objective To describe family cognition of breastfeeding and primiparas' breastfeeding behaviors 4 months postpartum and to explore the relationship between family cognition and new mother' breastfeeding behaviors. Methods A cross-sectional questionnaire survey was conducted to 200 primiparas at outpatient clinic in Beijing in terms of their breastfeeding behaviors and family cognition of breastfeeding. Results The rote of entire breastfeeding was 47.0％ ;and the score of family cognition was (23. 13 ±3. 19), which was at a moderate level. The total score of family cognition of primiparas who gave entire breastfeeding was significantly higher than those gave mixed or artificial feeding (F=8.26, df=2, P＜0. 01). Conclusions The breasffeeding rate in primiparas was relatively low,primipaaras who insists on giving entire breastfeeding have high level of family cognition. To help people form correct and active family cognition of breastfeeding and improve breastfeeding behavior in primiparas, health education should be launched toward not only primiparas but also their family numbers, especially the main caregiver.

To investigate the influence of Anxin granules combined with tirofiban on acute myocardial infarction (AMI) Patients after elective percutaneous coronary intervention (PCI). One hundred and twenty AMI patients were randomly divided into treatment group and control group. The patients in the two groups were all given Tirofiban 30mins before PCI . The treatment group was added Anxin granules 30 mins before and after PCI. Tissue factor (TF) and von willebrand factor (vWF) were tested at 6 hours after operation. Syndromatology alteration of traditional Chinese medicine (TCM) and bleeding complications were observed at 4 weeks after operation. Both TF and vWF at 6 hours after operation of the treatment group was lower than the control group significantly (P < 0.01), while the condition of myocardial ischemia at 90 mins after operation of the treatment group was better than control group with significance. The syndromatology alteration of TCM especially spontaneous perspiration and hypodynamia of the treatment group were improved significantly compared to control group 4 weeks after operation. All patients in both groups had no bleeding complications and thrombopenia. The study suggests that Anxin granules combined with tirofiba can improve the clinical efficacy and the endothelial function of AMI patients after PCI with no increase in bleeding events.
Aged ; Angioplasty, Balloon, Coronary ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Middle Aged ; Myocardial Infarction ; complications ; metabolism ; surgery ; Postoperative Hemorrhage ; drug therapy ; etiology ; metabolism ; prevention & control ; Thromboplastin ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo investigate the value of application of mandibular outer cortex as bone graft by comparing its bone absorption with cranial outer cortex.

METHODS8 minitype grown-up pigs at the age of 8 - 12 months underwent surgery of taking out the same size (2.5 cm x 1.0 cm) of outer cortex from mandible and craninium. The volume of the outer cortex was measured by volume-displacement method. Then the outer cortex of mandible and cranium were onlay grafted to the each side of the pig snout, respectively. 12 weeks later, 2 pigs were randomly selected for histological examination. The other 6 pigs were killed 24 weeks after surgery for measurement of the bone graft volume and histologic examination.

RESULTSThe bone graft absorption rate was (41 +/- 5)% for mandibular outer cortex and (46 +/- 12)% for cranial outer cortex, showing no significant difference between them (P = 0.51). The histologic examination results also had no marked difference in the bony healing and reforming between the two graft.

CONCLUSIONSMandibular outer cortex is a good donor site for onlay bone graft in craniofacial region.

Animals ; Bone Plates ; Bone Regeneration ; Bone Transplantation ; methods ; Female ; Male ; Mandible ; transplantation ; Skull ; transplantation ; Swine ; Swine, Miniature

OBJECTIVE: To study the effect of dhfr gene overexpression on ethanol-induced abnormal cardiac and vascular development in zebrafish embryos and underlying mechanisms. METHODS: dhfr mRNA was transcribed in vitro and microinjected into zebrafish fertilized eggs to induce the overexpression of dhfr gene, and the efficiency of overexpression was verified. Wild-type zebrafish were divided into a control group, an ethanol group, and an ethanol+dhfr overexpression group (microinjection of 6 nL dhfr mRNA). The embryonic development was observed for each group. The transgenic zebrafish Tg (cmlc2:mcherry) with heart-specific red fluorescence was used to observe atrial and ventricular development. Fluorescence microscopy was performed to observe the development of cardiac outflow tract and blood vessels. Heart rate and ventricular shortening fraction were used to assess cardiac function. Gene probes were constructed, and embryo in situ hybridization and real-time PCR were used to measure the expression of nkx2.5, tbx1, and flk-1 in the embryo. RESULTS: Compared with the ethanol group, the ethanol+dhfr overexpression group had a significant reduction in the percentage of abnormal embryonic development and a significant increase in the percentage of embryonic survival (P<0.05), with significant improvements in the abnormalities of the atrium, ventricle, outflow tract, and blood vessels and cardiac function. Compared with the control group, the ethanol group had significant reductions in the expression of nkx2.5, tbx1, and flk-1 (P<0.05), and compared with the ethanol group, the ethanol+dhfr overexpression group had significant increases in the expression of nkx2.5, tbx1, and flk-1 (P<0.05), which were still lower than their expression in the control group. CONCLUSIONS The overexpression of the dhfr gene can partially improve the abnormal development of embryonic heart and blood vessels induced by ethanol, possibly by upregulating the decreased expression of nkx2.5, tbx1, and flk-1 caused by ethanol.
Animals ; Ethanol ; Gene Expression Regulation, Developmental ; Heart ; Heart Ventricles ; Zebrafish ; Zebrafish Proteins