ABSTRACT
Tanshinones are bioactive ingredients derived from the herbal plant Salvia miltiorrhiza and are used for treating diseases of the heart and brain, thus ensuring quality of S. miltiorrhiza is paramount. Applying the endophytic fungus Trichoderma atroviride D16 can significantly increase the content of tanshinones in S. miltiorrhiza, but the potential mechanism remains unknown. In the present study, the colonization of D16 effectively enhanced the levels of Ca2+ and H2O2 in the roots of S. miltiorrhiza, which is positively correlated with increased tanshinones accumulation. Further experiments found that the treatment of plantlets with Ca2+ channel blocker (LaCl3) or H2O2 scavenger (DMTU) blocked D16-promoted tanshinones production. LaCl3 suppressed not only the D16-induced tanshinones accumulation but also the induced Ca2+ and H2O2 generation; nevertheless, DMTU did not significantly inhibit the induced Ca2+ biosynthesis, implying that Ca2+ acted upstream in H2O2 production. These results were confirmed by observations that S. miltiorrhiza treated with D16, CaCl2, and D16+LaCl3 exhibit H2O2 accumulation and influx in the roots. Moreover, H2O2 as a downstream signal of Ca2+ is involved in D16 enhanced tanshinones synthesis by inducing the expression of genes related to the biosynthesis of tanshinones, such as DXR, HMGR, GGPPS, CPS, KSL and CYP76AH1 genes. Transcriptomic analysis further supported that D16 activated the transcriptional responses related to Ca2+ and H2O2 production and tanshinones synthesis in S. miltiorrhiza seedlings. This is the first report that Ca2+ and H2O2 play important roles in regulating fungal-plant interactions thus improving the quality in the D16-S. miltiorrhiza system.
Subject(s)
Abietanes , Calcium , Endophytes , Hydrogen Peroxide , Plant Roots , Salvia miltiorrhiza , Salvia miltiorrhiza/metabolism , Salvia miltiorrhiza/microbiology , Hydrogen Peroxide/metabolism , Abietanes/biosynthesis , Abietanes/metabolism , Endophytes/metabolism , Endophytes/genetics , Plant Roots/microbiology , Plant Roots/metabolism , Calcium/metabolism , Calcium Signaling/drug effects , Lanthanum/pharmacology , Lanthanum/metabolism , Gene Expression Regulation, Plant , Hypocreales/metabolism , Hypocreales/geneticsABSTRACT
The seeds of Vitex negundo L. (Verbenaceae) have been commonly used as a folk remedy for the treatment of rheumatism and joint inflammation in Traditional Chinese Medicine. This study aimed to evaluate the anti-arthritic activity of the extract of V. negundo seeds (EVNS) using Freund's complete adjuvant (CFA) induced arthritis (AA) in rat model. As a result, EVNS, with abundant phenylnaphthalene-type lignans, significantly inhibited the paw edema, decreased the arthritis score and spleen index, and reversed the weight loss of CFA-injected rats. Histopathological studies showed a marked decrease of synovial inflammatory infiltration and synovial lining hyperplasia in the joints of EVNS-treated animals. The remarkable decrement of serum inflammatory factors (TNF-α, IL-1ß and IL-6) were observed in EVNS-treated rats, whereas, IL-10, an anti-inflammatory cytokine, was found to be significantly increased by EVNS. The expressions of COX-2 and 5-LOX in PBMC were also inhibited by administration of EVNS. Our results demonstrated that V. negundo seeds possessed potential therapeutic effect on adjuvant induced arthritis in rats by decreasing the levels of TNF-α, IL-1ß and IL-6 and increasing that of IL-10 in serum as well as down-regulating the levels of COX-2 and 5-LOX, and therefore may be an effective cure for the treatment of human rheumatoid arthritis.
Subject(s)
Anti-Inflammatory Agents/therapeutic use , Arthritis, Experimental/drug therapy , Inflammation Mediators/blood , Inflammation/drug therapy , Phytotherapy , Plant Extracts/therapeutic use , Vitex/chemistry , Animals , Anti-Inflammatory Agents/pharmacology , Antirheumatic Agents/pharmacology , Antirheumatic Agents/therapeutic use , Arthritis, Experimental/blood , Arthritis, Experimental/pathology , Arthritis, Rheumatoid/drug therapy , Cyclooxygenase 2/blood , Down-Regulation , Edema , Freund's Adjuvant , Inflammation/blood , Inflammation/chemically induced , Interleukins/blood , Joints/drug effects , Joints/pathology , Leukocytes, Mononuclear/drug effects , Lignans/pharmacology , Lignans/therapeutic use , Lipoxygenases/blood , Male , Naphthalenes/pharmacology , Naphthalenes/therapeutic use , Plant Extracts/pharmacology , Plant Extracts/standards , Rats, Wistar , Seeds , Synovial Membrane/drug effects , Synovial Membrane/pathology , Tumor Necrosis Factor-alpha/blood , Weight Loss/drug effectsABSTRACT
During the course of paternity test, three samples in two cases were apparently homozygous at the CSF1PO locus using AmpFlSTRs Identifiler PCR Amplification kits, but using the PowerPlexs 16 kit, the three individuals were found to be heterozygous. This puzzling problem was solved by using multiple analytical approaches, including the use of different primer pairs and the characterization of the mutation causing the ''null allele.'' Dropout was caused by a single mutation event in the presumptive binding site of the forward primer. While the frequency of these silent alleles remains low (0.5% in our study), it is suggested that appropriate measures should be taken for database comparisons and that allelic dropout should be further investigated by sequence analysis and be reported to the forensic community.
Subject(s)
DNA Primers , Mutation , Alleles , Binding Sites/genetics , China , Homozygote , Humans , Paternity , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
Numerous candidate genes have been proposed as susceptibility factors for the development of esophageal squamous cell carcinoma (ESCC). XRCC1 (X-ray cross-complementing group 1) codon 280 and ERCC2 (excision repair cross complementing group 2) codon 751 polymorphisms were studied in ESCC in a Chinese population. The aim of this study is to investigate the potential association between single-nucleotide polymorphisms (SNP) of XRCC1 codon 280 His and ERCC2 codon 751 Gln polymorphisms and ESCC. Peripheral blood samples of 200 cases and 200 age-and-gender matching controls were collected from a Chinese population and the two polymorphisms were studied by means of polymerase chain reaction (PCR) restriction fragment length polymorphism techniques. Our results showed that XRCC1 codon 280 His allele had no significant difference between ESCC patients and normal controls (P > 0.05), while ERCC2 codon 751Gln allele was associated with a borderline decrease of ESCC (odds ratio [OR] = 0.628, 95% confidence interval [CI]: 0.400-0.986).
Subject(s)
Carcinoma, Squamous Cell/genetics , DNA-Binding Proteins/genetics , Esophageal Neoplasms/genetics , Polymorphism, Single Nucleotide/genetics , Xeroderma Pigmentosum Group D Protein/genetics , Alleles , Asian People/ethnology , Carcinoma, Squamous Cell/ethnology , Case-Control Studies , Codon/genetics , DNA Repair/genetics , Esophageal Neoplasms/ethnology , Female , Genotype , Humans , Male , Middle Aged , X-ray Repair Cross Complementing Protein 1ABSTRACT
<p><b>OBJECTIVE</b>To report the results of curative and adverse effects of compound huangdai tablet (CHDT) as induction therapy for 193 patients with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>CHDT was administered 1.25 g orally three times a day after meal for three days, then the dosage was gradually increased to 7.5 g/d.</p><p><b>RESULTS</b>One hundred and ninety-three patients achieved complete remission (CR), 78.8% of whom in 30 to 60 days with an average time of 44.3 d. No serious infection, bleeding or DIC occurred during the treatment course. The major adverse effects were gastrointestinal symptoms. There was no change in lanine transaminase, urea, creatinine or electrocardiographic QTc interval in 110 APL patients observed before and after the treatment.</p><p><b>CONCLUSION</b>CHDT therapy is a modality of higher CR rate, good safety and tolerance without bone marrow suppression for APL patients.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Follow-Up Studies , Leukemia, Promyelocytic, Acute , Drug Therapy , Phytotherapy , Plant Preparations , Therapeutic Uses , Treatment OutcomeABSTRACT
OBJECTIVE: To study the application of PCR-SSCP in forensic mtDNA typing. METHODS: Primers flanking the mtDNA HV-I and HV-II regions were designed. By PCR-SSCP techniques, 70 family trios and 140 unrelated Wuhan Han individuals were investigated and analyzed. RESULTS: In 70 family trios, the SSCP profiles in region HV-I and HV-II of children were not same to that of their fathers in 98.57% and 97.13% respectively but were identical with their mothers. In 140 unrelated Wuhan Han individuals, 21 haplotypes were found in HVI, GD = 0.9556; 16 haplotypes were found in HVII, GD = 0.9356. CONCLUSION: PCR-SSCP technique may be useful in forensic mtDNA typing, especially for screening the suspects.
Subject(s)
DNA Fingerprinting/methods , DNA, Mitochondrial/genetics , Haplotypes , Polymerase Chain Reaction/methods , Polymorphism, Single-Stranded Conformational , DNA Primers , DNA, Mitochondrial/blood , Forensic Genetics/methods , Humans , Pedigree , Sequence Analysis, DNAABSTRACT
OBJECTIVE: To investigate the polymorphism of DXYS267 locus in China Han population and find the application and characters of its Y-specific single nucleotide substitutions. METHODS: The locus was analyzed by PCR and PAGE in silver-staining. The Y-specific STR was amplified with newly designed primers according to the Y-specific single nucleotide substitutions. RESULTS: Six alleles were detected in Han population in Wuhan. Exact tests demonstrated that genotype frequencies did not deviate from Hardy-Weinberg equilibrium. Heterozygosity of DXYS267 was 0.6706, discrimination power (DP) was 0.8433, and the probability of paternity exclusion (PE) was 0.5957. The Y-specific STR of DXYS267 was successfully amplified with the new primer. The 4 alleles for Y-STR were detected with haplotype diversity (HD) 0.6372. CONCLUSION: The DXYS267 locus is appropriate for individual identification and paternity testing. The new primer is useful for individual and paternity testing involving brothers and mixed stains.
