OBJECTIVE: To explore the feasibility of biological method to identify the biological attribute of samples at crime scene. METHODS: Thirty samples of ten blood stains, ten saliva stains and ten semen stains were selected, and all the samples were processed by the routine method and biomolecular method, respectively. Both RNA and DNA were isolated using DNA-RNA co-extraction technology and the mRNA was converted into cDNA using reverse transcription PCR (RT-PCR). Three pairs of specific primers were designed for blood stain, saliva stain and semen stain based on the different target genes in three specific tissues and the primers were amplified using real-time fluorescent quantitative PCR. The differences in these biological samples were evaluated by melting temperature (Tm) values and the size of the amplification fragment. RESULTS: The Tm values of blood stain, saliva stain and semen stain were (84.5 +/- 0.2) degrees C, (76.9 +/- 0.3) degrees C and (88.5 +/- 0.2) degrees C, respectively. The length of PCR fragments of them was 177bp, 134bp and 294bp, respectively. CONCLUSION Compared with the routine method, RT-PCR with real-time fluorescent quantitative PCR is highly specific, sensitive and reliable to identify the biological attribute of evidence, and can be potentially applied to determine evidence attribute in forensic practice.
Blood Stains ; DNA/isolation & purification* ; DNA Primers ; Forensic Medicine/methods* ; Genetic Markers ; Humans ; Male ; Polymerase Chain Reaction/methods* ; RNA/isolation & purification* ; RNA, Messenger/analysis* ; Saliva ; Semen ; Sensitivity and Specificity

PURPOSE: Recently, COMMD1 has been identified as a novel interactor and regulator of hypoxia-inducible factor-1 and nuclear factor kappa B transcriptional activity. The goal of this study was to determine the difference of COMMD1 expression in the placentas of women with normal and preeclamptic (PE) pregnancies. MATERIALS AND METHODS: Immnoperoxidase and immunofluorescent staining for COMMD1 was performed on nine normal and nine severe PE placental tissues, and COMMD1 mRNA expression was quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: The expression of mRNA of COMMD1 was significantly higher in the study group than in the control group. The immunoreactivity was higher especially in the syncytiotrophoblast of PE placentas than in the control group. CONCLUSION: This study demonstrated increased placental COMMD1 expression in women with severe preeclampsia compared to that found in women with normal pregnancies, and this finding might contribute to a better understanding of the pathophysiology of preeclampsia.
Adaptor Proteins, Signal Transducing/genetics/isolation & purification/*metabolism ; Adult ; Female ; Humans ; Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy ; RNA, Messenger/metabolism

OBJECTIVETo investigate the effect of cryptoporus polysaccharide(CP)on lipopolysaccharide(LPS)-induced production of monocyte chemoattractant protein-1(MCP-1)in human lung epithelial A549 cells.

METHODSA549 cells were stimulated with LPS in the presence or absence of CP. The protein concentration and mRNA expression of MCP-1 were determined by enzyme-linked-immunosobent assay(ELISA)and semi-quantitative RT-PCR, respectively.

RESULTThe protein concentration of MCP-1 was significantly increased by LPS 1000 microg/L at 24 h. There were no effects on the growth and viability of A549 cells in the presence of CP 100 microg/L or dexamethasone 1 mumol/L. However, CP 100 microg/L or dexamethasone 1 micromol/L significantly inhibited the protein concentration and mRNA expression of MCP-1 induced by LPS.

CONCLUSIONCP can regulate MCP-1 production, which may be associated with its effects on lung inflammation.

Cell Line ; Chemokine CCL2 ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Polyporaceae ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism

OBJECTIVETo explore the immuno-neurologic regulation of hypertension and its inherent law as well as the mechanisms of curing and systemic regulating effect of ZiShuiJiangHuoYin(ZSJHY).

METHODTo detect expression level of AT-1mRNA in two-kidney-one-clamp renal hypertension rat lymphocyte cell by means of RT-PCR.

RESULTThe level of AT-1 mRNA in lymphocyte was higher in group of 2K1C-RHR than that in normal group. ZSJHY (20 g/kg, 40 g/kg) had regulating effect on this.

CONCLUSIONDepressing excessive expression of lymphocyte AT-1 mRNA may be one of the mechanisms where ZSJHY exert immunoregulation action.

Animals ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hypertension, Renovascular ; metabolism ; Lymphocytes ; metabolism ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Angiotensin ; biosynthesis ; genetics

OBJECTIVE: To explore the expression level of class I integrase (intI 1) mRNA in Acinetobacter baumannii from biofilm cells and planktonic cultured cells ,and to analyze the drug-resistance of Class I integron positive strains. METHODS: Acinetobacter baumannii were collected from hospitals,and Class I integron strains were screened by gene amplification. Total RNA of Class I integron positive strains was extracted, and the intI1 mRNA expression in the bioflim cells and planktonic cultured cells was measured by RT-PCR. Susceptibilities to antibiotics of Class I integron positive strains were also examined. RESULTS: The intI1 gene mRNA was expressed under 2 conditions, and the mRNA expressed in the biofilm cells was about 4 times higher than that in the planktonic cultured cells. Among the 64 strains of Acinetobacter baumannii, 46 strains were Class I integron positive strains. The antibiotic resistance of intI1 gene cassette-positive strains was higher than that of gene cassette-negative strains. CONCLUSION The intI1 gene mRNA can be up-regulated in Acinetobacter baumannii biofilm cells.Class I integron plays an important role in drug resistance. It is much easier to capture gene cassettes for bacteria under biofilm condition.
Acinetobacter Infections ; microbiology ; Acinetobacter baumannii ; genetics ; isolation & purification ; Base Sequence ; Biofilms ; Drug Resistance, Multiple ; genetics ; Humans ; Integrases ; biosynthesis ; genetics ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo explore the effect of Ray cartilage glycosaminoglycans (RCG) on the expression of MMP-9 in Lewis lung carcinoma of mice.

METHODThe model of mice with Lewis lung carcinoma was induced. The experimental mice were randomly divided into normal saline group, RCG groups at varied concentrations and CTX group. Tumor growth state was observed, and tumor inhibitory rate of primary tumor and number of lung metastasis focus were measured. The expression of MMP-9 mRNA and protein in Lewis lung carcinoma was determined with RT-PCR and Western blot.

RESULTAs compared with normal saline group, tumor growth curves in RCG groups were smooth, there were significant differences of inhibitory rates of primary tumor and number of lung metastasis focus between RCG groups and normal saline group, and MMP-9 mRNA and protein expression levels in RCG groups were reduced significantly.

CONCLUSIONRCG can inhibit effectively the growth and metastasis of implanted Lewis lung carcinoma in C57BL/6 mice, which is probably attributed to reducing the expression of MMP-9 mRNA and protein.

Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Lewis Lung ; enzymology ; pathology ; Cartilage ; chemistry ; Cell Line, Tumor ; Glycosaminoglycans ; isolation & purification ; pharmacology ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Skates (Fish)

OBJECTIVETo investigate the effects of high glucose on expression of matrix metalloproteinase-2 (MMP-2) and extracellular matrix metalloproteinase inducer (EMMPRIN) in human umbilical vein endothelial cells (HUVECs).

METHODSHUVECs were cultured in DMEM media containing high glucose with or without polyphenols for 24 hours respectively. The activity of MMP-2 in the supernatant was detected by zymography. The expression of MMP-2 mRNA and protein in HUVECs were detected by RT-PCR and Western blot respectively. The expression of EMMPRIN mRNA and protein in the cells were determined by RT-PCR as well as immunocytochemistry and Western blot respectively.

RESULTSThe expression of MMP-2 and EMMPRIN mRNA were suppressed by a high concentration of glucose. Both the MMP-2 activity and protein level of EMMPRIN expression were also significantly decreased. Polyphenols abolished all the above changes of HUVECs induced by a high glucose level (P < 0.05).

CONCLUSIONSAn acute high glucose stimulation down-regulates the activity of MMP-2, the expressions of MMP-2 and EMMPRIN at RNA and protein levels in the endothelial cells, which may play an important roles in diabetic vascular complications in the early phase. Polyphenols treatment can diminish the detrimental effects of high glucose on HUVECs.

Basigin ; genetics ; metabolism ; Cells, Cultured ; Endothelial Cells ; metabolism ; Flavonoids ; isolation & purification ; pharmacology ; Glucose ; metabolism ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Phenols ; isolation & purification ; pharmacology ; Polyphenols ; RNA, Messenger ; metabolism ; Tea ; chemistry ; Umbilical Veins ; cytology

AIMTo determine the effect of lysophosphatidylcholine (LPC) on the expression of vascular endothelial growth factor (VEGF) in human umbilical veins endothelial cell line (ECV304) and the inhibitory effect of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucoside (ST I) in vitro.

METHODSExposure to 2.5 mg x L(-1) LPC or LPC + ST I for 24 hours, VEGF protein was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile, VEGF mRNA expression in ECV304 was examined by in situ hybridization. VEGF165 mRNA was examined by RT-PCR and Realtime RT-PCR.

RESULTSLPC upregulated VEGF protein and VEGF mRNA expression in the ECV304 cells. ST I was shown to markedly inhibit the LPC-induced increase of VEGF protein and VEGF165 mRNA (P < 0.001).

CONCLUSIONLPC can induce a strong expression of VEGF in ECV304 cells and ST I can inhibit it.

Cells, Cultured ; Endothelial Cells ; metabolism ; Glucosides ; isolation & purification ; pharmacology ; Humans ; Lysophosphatidylcholines ; antagonists & inhibitors ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Stilbenes ; isolation & purification ; pharmacology ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

AIMTo investigate the protective effects of shark hepatic stimulator substance (sHSS) against acute hepatic injury induced by acetaminophen (AAP) in mice.

METHODSAcute hepatic injury model of Balb/c mice was induced by a single intraperitoneal injection of AAP (200 mg.kg-1, i.p.). Serum ALT and AST activities were analyzed. The changes of microstructure and ultrastructure of hepatocyte were observed under optical and electronic microscope. The hepatocyte apoptosis was analyzed by flow cytometer and the expression level of Fas mRNA was determined by RT-PCR.

RESULTSThe activities of serum ALT and AST were significantly decreased and both necrosis and inflammatory infiltration were improved in the mice treated with sHSS 3.0 and 1.5 mg.kg-1. sHSS (3.0 mg.kg-1) prevented the ultrastructural changes of hepatocytes caused by AAP, decreased the percentage of apoptotic cells, and downregulated the expression level of Fas mRNA.

CONCLUSIONsHSS protected hepatocytes from AAP-induced injury, which might be associated with its protection of the mitochondria and inhibition of apoptosis and expression of Fas mRNA in hepatocytes.

Acetaminophen ; Animals ; Apoptosis ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; pathology ; Female ; Growth Substances ; isolation & purification ; pharmacology ; Mice ; Mice, Inbred BALB C ; Peptides ; isolation & purification ; pharmacology ; Protective Agents ; pharmacology ; RNA, Messenger ; genetics ; Random Allocation ; Sharks ; fas Receptor ; biosynthesis ; genetics

This study is purposed to investigate the effects of praeruptorin A (PA) and praeruptorin C (PC) on UGT1A1 in HepG2 cells through hCAR pathway. PA and PC were incubated with HepG2 cells for 24 h and 48 h, mRNA and protein expressions of UGT1A1 were determined by real-time PCR and Western blotting assays. Additionally, effects of PA and PC on UGT1A1 mRNA and protein expressions were also measured after transient transfection of a specific CAR siRNA for 72 h in HepG2 cells. UGT1A1 mRNA and protein expression levels were significantly increased by PA and PC after incubation for 48 h. Moreover, the mRNA and protein up-regulations of UGT1A1 were attenuated by transient transfection of a specific CAR siRNA, suggesting the induction was mediated by CAR. The results suggest that PA and PC can significantly up-regulate UGT1A1 expression partially via the CAR-mediated pathway.
Apiaceae ; chemistry ; Coumarins ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Glucuronosyltransferase ; genetics ; metabolism ; Hep G2 Cells ; Humans ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; Signal Transduction ; Transfection