Objective To analyze the effect of eukaryotic plasmids containing wild (sense) or anti- sense methylenetetrahydrofolate reductase (MTHFR) gene on cell viability and transcription level of tumor related genes in human gastric cancer cell line.Methods Human gastric cancer cell line MKN-45 was cultured.Recombinant plasmids containing wild MTHFR (W) or antisense MTHFR (A) gene, pCMV-W and pCMV-A,were constructed.Then pCMV-W,pCMV-A and pCMV blank plasmid were transfected into MKN45 cells respectively by using lipofect.Cell viability was analyzed by 3-(4,5-bime- thylthiazolyl-2)-2,5-diphenyhetrazolium dromide(MTT).The transcription levels of Dnmt 1,c-myc, p21~(WAF1) and hMLH1 genes were detected by real-time polymerase chain reaction(PCR).Results Cell vi- ability remarkably increased in those transfected with wild MTHFR (P＜0.01),which was contrary to those transfected with antisense MTHFR(P＜0.01).The expression of those tumor related genes mRNAs were all remarkably decreased in the MKN45-W cells in comparison with those in the MKN45-pCMV cells.No significant difference in the expressions of those tumor related genes mRNAs were found between the MKN45 cells transfected with pCMV-A and blank pCMV.Conclusion MTHFR influences cell viability and the expres- sion level of tumor related genes in human gastric cancer cell line MKN45.

OBJECTIVETo study the effects of extremely low frequency electromagnetic fields (ELF EMFs) on apoptosis and cell cycle of mouse brain and liver cells.

METHODSMice were exposed to 50 Hz, 0.2 mT or 6.0 mT electromagnetic fields for 2 weeks. TUNEL and flow cytometric methods were used to analyze apoptosis and cell cycle of brain and liver cells.

RESULTSAfter exposure to 0.2 mT and 6.0 mT ELF EMFs for 2 weeks, apoptosis rates of brain cells [(5.60 +/- 1.47)% and (4.73 +/- 0.48)% respectively] were higher than that of control [(2.90 +/- 0.75)%], and apoptosis rates of liver cells [(4.19 +/- 2.08)% and (3.38 +/- 0.65)% respectively] were higher than that of control [(1.84 +/- 0.76)%]. G0/G1 cell percentage of brain cells [(80.21 +/- 1.68)% and (79.54 +/- 0.56)% respectively] were higher than that of control [(76.85 +/- 0.83)%], and those of liver cells [(79.42 +/- 1.80)% and (80.47 +/- 1.79)% respectively] were higher than that of control [(73.36 +/- 3.10)%]. The above differences were all statistically significant as P < 0.05. At the same time S and G2 + M cell percentage of brain and liver cells were significantly decreased.

CONCLUSIONExposure to 50 Hz EMFs may alter cell cycle and induce apoptosis of mouse brain and liver cells.

Animals ; Apoptosis ; radiation effects ; Brain ; cytology ; radiation effects ; Cell Cycle ; radiation effects ; Electromagnetic Fields ; Flow Cytometry ; In Situ Nick-End Labeling ; Liver ; cytology ; radiation effects ; Male ; Mice

OBJECTIVETo evaluate the effect of PKC-delta inhibitor Rottlerin on human colon cancer cells and its mechanism.

METHODSHuman colon cancer cell line SW1116 cells were treated with Rottlerin. The transcriptional level of DNA methyltransferase (Dnmt)1, Dnmt3a and Dnmt3b was detected by real-time RT-PCR. Cell cycle distribution was evaluated by flow cytometry (FCM). In addition, cellular morphological changes were examined by light microscopy.

RESULTSPKC-delta inhibitor decreased the expression of Dnmt1, Dnmt3a mRNA, up-regulated APC, p21(WAF1) and p16(INK4A) mRNA. Demonstarted by flow cytometry, Rottlerin increased the percentage of cell cycle G0/G1 phase cell numbers (P = 0.02) and decreased the percentage of cell cycle G2/M phase cell numbers (P = 0.01). Remarkable changes of cellular morphology were observed under light microscope: The volume and cytoplasm of cells treated with Rottlerin were increased. The cell contour was not very clear, and mitotic figures were less frequently seen.

CONCLUSIONPKC-delta inhibitor Rottlerin inhibites cell division and proliferation of the colon cancer SW1116 cells through regulating DNA methylation and blocking the signaling pathway of mitogen-activated protein kinase (MAPK).

Acetophenones ; pharmacology ; Adenomatous Polyposis Coli Protein ; genetics ; Benzopyrans ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Protein Kinase C-delta ; antagonists & inhibitors ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects

OBJECTIVETo analyze the specificity, sensitivity and receiver operating characteristic (ROC) curve of plasma elafin for diagnosis of skin acute graft-versus-host disease (aGVHD), and to explore its clinical diagnostic value.

METHODSIncidence of skin aGVHD from fifty-three patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) were observed prospectively in Guangdong General Hospital from Apr 2010 to Aug 2011. The plasma concentrations of elafin were detected by enzyme-linked immunosorbent assay (ELISA). Skin biopsies were taken from 28 patients with skin rash, and elafin expression in the skin was detected by immunohistochemistry. Positive expression was defined as significant staining of at 50% of the depth of the epidermis, excluding the granular cell layer and the acrosyringium.

RESULTSAmong 28 patients with skin rash, twenty-five were considered as skin aGVHD by clinical diagnosis, seventeen were confirmed as skin aGVHD by pathological biopsy. 11 cases were elafin positive by immunohistochemical staining. Elafin protein was overexpressed in aGVHD skin tissue (P = 0.001). Plasma concentrations of elafin were significantly higher in patients with skin aGVHD (positive) group than in those without skin aGVHD (negative) group (P = 0.005), among which there being no statistically significant difference in plasma elafin level between patients with grade I skin aGVHD group and negative group(P = 0.971), but being statistically significant difference compared patients with grade II-IV skin aGVHD group with those with grade I skin aGVHD group (P = 0.02) and with negative group (P = 0.008). Using the pathological diagnosis as the gold standard, the estimated specificity and the sensitivity of clinical diagnosis criteria were 27.3% and 100%, respectively, and those of tissue elafin protein level were 100% and 64.7%, respectively. The area under the ROC curve was 0.909 (0.797 - 1.021) when plasma concentrations of elafin was used in diagnosis of skin aGVHD. The sensitivity was 82.4% and the specificity was 81.8 % when the critical value was set at 1456.043 µg/L.

CONCLUSIONPlasma concentration of elafin is significantly higher at the onset of skin aGVHD. It can be used as biochemical marker of skin aGVHD and has higher value in diagnosis of skin aGVHD.

Adolescent ; Adult ; Elafin ; blood ; Female ; Graft vs Host Disease ; blood ; diagnosis ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; ROC Curve ; Sensitivity and Specificity ; Skin Diseases ; blood ; diagnosis ; etiology ; Young Adult

This study was purposed to investigate the efficacy and safety of intravenous injecting itraconazole (ITCZ) as empirical antifungal therapy in the patients with hematological malignancies. According to recommendation in IDSA guidebook, the patients suffered from fever during neutropenia and inefficacy of treatment using broad-spectrum antibiotics for 4 days should receive intravenous injection of ITCZ as empirical antifungal therapy. The results showed that the overall clinical response rate to ITCZ injection was 62.9% (22/35), and the success rate of achieving composite endpoints was 54.3% (19/35). Mild adverse reactions were observed in 6 patients (17.1%). The injection of ITCZ was stopped in 2 patents (5.7%) due to adverse reaction. Further analysis revealed that the response rate was higher in patients with fever prior to the start of ITCZ within five days than beyond five days (P = 0.031). The response rate was higher in patients with possible invasive fungus infection (IFI) than that in patients with probable and confirmed IFI (P = 0.002). The prophylactic antifungal treatment during neutropenia displayed no significant influence on efficacy of empirical antifungal therapy with itraconazole (P = 0.054). It is concluded that the good efficacy and safety of empirical ITCZ injection for hematological malignancies patients is efficient and safe.
Adolescent ; Adult ; Aged ; Antifungal Agents ; administration & dosage ; therapeutic use ; Female ; Hematologic Neoplasms ; drug therapy ; Humans ; Injections, Intravenous ; Itraconazole ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

The aim of this study was to investigate the renal function in 149 patients receiving myeloablative allogeneic hematopoietic stem cell transplantation (allo-HSCT) from June 2005 to June 2010 in our hospital, and analyze the risk factors resulting in kidney insufficiency and experience in diagnose and therapy. The creatinine clearance (CrCL) and serial creatinine level were evaluated before and after allo-HSCT within 100 days and 1 year. Non-radiation conditioning regimens were used for any patients. The acute kidney insufficiency (AKI) was defined as at least a 1.5-fold rise in serum creatinine level after allo-HSCT within the first 100 days. The chronic kidney insufficiency (CKI) was defined as the creatinine clearance < basal level within 3 months to 1 year after allo-HSCT. The results showed that the kidney insufficiency was found in 41 patients, in which the incidence of AKI was 32/149 (21.5%). CsA, amphotericin B (P = 0.025) and ES (P = 0.022) were defined as risk factors for AKI. The incidence of CKI was 18/138 (13%). cGVHD (P = 0.013) and TA-TMA (P = 0.012) were associated with the development of CKI. The 2-year survival was lower in patients with kidney dysfunction than that in patients without kidney dysfunction (39% vs 74.1%, P < 0.001). The main factors resulting in kidney insufficiency were defined as infection (52%), GVHD (20%), TA-TMA (12%) and tumor relapse (12%). It is concluded that kidney insufficiency is an important complication of allo-HSCT. Careful monitoring kidney function, minimizing the use of amphotericin B, prophylaxis and effective treatment of fungal infection, GVHD and TA-TMA may be effective preventive measures to decrease the incidence of kidney insufficiency.
Acute Kidney Injury ; etiology ; Adolescent ; Adult ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Renal Insufficiency ; etiology ; Retrospective Studies ; Risk Factors ; Transplantation, Homologous ; Young Adult

ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.
Apocynaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Flowers ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control

Objective:To study the predictive value of carotid plaque score for evaluating the severity of coronary artery lesions.Methods:A total of 152 patients with coronary angiography diagnosed coronary artery disease (CAD) in our hospital from 2016-01 to 2016-05 were selected.According to coronary branch involvement,the patients were divided into 3 groups:Single branch lesion group,n=72,Double branch lesion group,n=38 and Multi branch lesion group,n=42.The basic conditions as plaque detection rate,high-density lipoprotein cholesterol (HDL-C),body mass index (BMI) were collected and carotid ultrasound examination was conducted in all patients.Carotid plaque condition and coronary lesion degree were calculated by semi-quantitative score and Gensini score methods;the relationship between semi-quantitative score and Gensini score was compared.Results:Total cholesterol (TC),HDL-C,LDL-C and BMI were similar among different groups,P＞0.05.Compared with Single branch lesion group,Double branch lesion group and Multi branch lesion group had more smokers and the higher plaque detection rate,P＜0.05.Semi-quantitative score and Gensini score were elevating as coronary lesion branch increasing accordingly.Conclusion:The severity of coronary lesions could be evaluated and predicted by semi-quantitative calculation of carotid integral.

OBJECTIVETo investigate the effects of extremely low frequency electromagnetic fields (ELF EMFs) on male reproduction in mice.

METHODS94 adult male mice were exposed to 50 Hz sinusoidal electromagnetic fields of 0.2, 3.2 or 6.4 mT for 2 weeks or 4 weeks. Testicular histology and weight, sperm amount, sperm motility and morphology were measured. The percentages of different ploidy cells and cell phases, and DNA content of testis cells were estimated by flow cytometry. The micronucleus rate of bone-marrow cell was also observed.

RESULTSThe testicular weight of the mice exposed to 6.4 mT for 4 weeks [(76.06 +/- 32.25) mg] was significantly lower than that of the control [(111.44 +/- 19.99) mg, P < 0.05]; no significant histopathological changes were observed on the testis in EMFs exposed mice;the sperm amount was decreased after EMFs exposure for 4 weeks, and those of the mice exposed to 0.2 mT and 6.4 mT for 4 weeks [(4.87 +/- 0.94) x 10(6)/ml and (4.30 +/- 1.89) x 10(6)/ml respectively] were significantly lower than that of the control [(6.67 +/- 0.70) x 10(6)/ml, P < 0.05]; the rates of sperm motility also showed a decline. After 0.2, 3.2 or 6.4 mT EMFs exposure for 2 weeks, the deformity rates of sperm [(7.416 +/- 3.352)%, (6.862 +/- 2.947)% and (8.112 +/- 4.615)% respectively] were significantly higher than that of the control [(4.098 +/- 2.028)%, P < 0.01]. Similarly, those of the mice exposed for 4 weeks [(10.267 +/- 3.836)%, (11.027 +/- 7.059)%, (8.814 +/- 3.678)% respectively] were higher than that of the control [(3.714 +/- 1.830)%]. After 6.4 mT exposure for 2 weeks, the percentages of 1C testis cells [(69.56 +/- 4.07)%] was significantly lower than that of the control [(73.45 +/- 3.10)%, P < 0.05]. There were not any remarkable changes in those of 2C, 4C cells. DNA content in different ploidy cells of the mice exposed to 6.4 mT was decreased. Moreover, the cell percentage in S phase was increased significantly (P < 0.01).

CONCLUSIONELF EMFs exposure may have some adverse effects on reproduction in mice.

Animals ; DNA ; metabolism ; Electromagnetic Fields ; Male ; Mice ; Random Allocation ; Reproduction ; radiation effects ; Sperm Count ; Sperm Motility ; radiation effects ; Spermatozoa ; cytology ; metabolism ; radiation effects ; Testis ; cytology ; radiation effects

Matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF-IMS) has been a classical technique for studying proteomics in present and a tool for analyzing the distribution of proteins and small molecules within biological tissue sections. MALDI-TOF-IMS can analyze multiple unknown compounds in biological tissue sections simultaneously through a single measurement which can obtain molecule imaging of the tissue while maintaining the integrity of cellular and molecules in tissue. In recent years, imaging mass spectrometry technique develops relatively quickly in all biomedical domain. This paper based on the relevant data and reviews the present developing level of MALDI-TOF-IMS, the principle of imaging mass spectrometry, methology and the prospect in forensic pathology.
Diagnostic Imaging ; Forensic Sciences/methods* ; Humans ; Male ; Proteins ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization