The field of forensic DNA typing,often referred to as“DNA fingerprinting,”has evolved and expanded considerably since its beginnings in the mid-1980s.Originally,forensic DNA typing was primarily used for individual ...The field of forensic DNA typing,often referred to as“DNA fingerprinting,”has evolved and expanded considerably since its beginnings in the mid-1980s.Originally,forensic DNA typing was primarily used for individual identification and criminal investigations,but it has evolved into a versatile discipline with a wide range of applications.This article addresses the growing scope of forensic genetics,which includes advances in DNA sequencing technologies,mixture analysis,body fluid identification,phenotypic profiling,forensic genealogy,microbiological analysis,exploration of novel markers,and ethical and legal considerations.These developments have enabled the analysis of difficult samples and provided comprehensive insights into the origins of biological evidence.In an ever-evolving landscape,forensic genetics continues to shape the future of forensic science by providing new tools and techniques that help deliver justice in an increasingly complex world.展开更多
DNA molecules are green materials with great potential for high-density and long-term data storage.However,the current data-writing process of DNA data storage via DNA synthesis suffers from high costs and the product...DNA molecules are green materials with great potential for high-density and long-term data storage.However,the current data-writing process of DNA data storage via DNA synthesis suffers from high costs and the production of hazards,limiting its practical applications.Here,we developed a DNA movable-type storage system that can utilize DNA fragments pre-produced by cell factories for data writing.In this system,these pre-generated DNA fragments,referred to herein as“DNA movable types,”are used as basic writing units in a repetitive way.The process of data writing is achieved by the rapid assembly of these DNA movable types,thereby avoiding the costly and environmentally hazardous process of de novo DNA synthesis.With this system,we successfully encoded 24 bytes of digital information in DNA and read it back accurately by means of high-throughput sequencing and decoding,thereby demonstrating the feasibility of this system.Through its repetitive usage and biological assembly of DNA movable-type fragments,this system exhibits excellent potential for writing cost reduction,opening up a novel route toward an economical and sustainable digital data-storage technology.展开更多
Purpose: To alert the medical community that whole exome sequencing can find accessory gene changes in well-known syndromes that alter preventive health care and management. Meaning: A collagen type VI gene change add...Purpose: To alert the medical community that whole exome sequencing can find accessory gene changes in well-known syndromes that alter preventive health care and management. Meaning: A collagen type VI gene change adds muscle weakness, hypermobility, and dysautonomia concerns to usual management considerations for Down syndrome. Methods: Commercial whole exome sequencing combined with clinical interpretation of DNA sequence change added new considerations to patient management and parental counsel. Results: An 11-year-old child with the trisomy 21 form of Down syndrome who was evaluated for extraordinary joint laxity had a heterozygous collagen type VI aspartic to glutamic acid (COL6A3 c.6360 C>G p.Asp2120Glu) gene change found by whole exome sequencing. The DNA variant was qualified as having strong relevance to the enhanced hypermobility due to prior association of collagen 6 gene changes with myopathy. Conclusions: Dual diagnosis of Ehlers-Danlos syndrome was not assigned because the patient lacked criteria like bruising, unusual scars, or selected dysautonomia symptoms. The concept of a hypermobility spectrum offers advantages for management of its constituent conditions if clinically guided ascertainment and DNA diagnostics are employed.展开更多
s To investigate the DNA polymorphism of Sporothrix schenckii ( S schencki i ) and to find the relationship between DNA patterns and geographic areas and clinical manifestations Method The total DNA was ...s To investigate the DNA polymorphism of Sporothrix schenckii ( S schencki i ) and to find the relationship between DNA patterns and geographic areas and clinical manifestations Method The total DNA was extracted with hexadecyltrimethyl ammonium bromide Random A mplification of Polymorphic DNA (RAPD) assay was used to study DNA typing of 24 strains of S schenckii collected from different areas and isolated from di fferent clinical types Results Of seven random primers used, three primers (OPAA11, OPD18 and OPB07) gave good reactions, the sequences of which were 5' ACCCGACCTG 3', 5' GAGAGCCAAC 3', 5 ' GGTGAC^GCAG 3' respectively The RAPD patterns of the 24 isolates were not completely identical, showing certain degrees of hereditary variability Differ ent isolates showed a common conserved DNA band with the same primer Different clinical types showed different genotypes Conclusion RAPD analysis is useful in DNA typing of S schenckii , the DNA band type of which is related to geographic origin and Clinical manifestation展开更多
The binding between indirubin and calf thymus DNA in vitro has been verified by meansof the isotope labelling method, spectrophotometric method and thermal denaturation meas-urements. The λmax 207 nm of indirubin shi...The binding between indirubin and calf thymus DNA in vitro has been verified by meansof the isotope labelling method, spectrophotometric method and thermal denaturation meas-urements. The λmax 207 nm of indirubin shifted toward longer wave length with decrease ofabsorbance after the incubation of indirubin with DNA. The escalation of Tm value of DNAinduced by indirubin was about 2.4°C and it was reproducible. The binding force between themwas rather weak, as indirubin molecules were easily released during the precipitation withalcohol or the gel filtration. The binding was not affected by sodium chloride even at high con-centration but greatly decreased (to 20-30% of the control) in the presence of 8 M urea.These results showed that the binding between indirubin and DNA might be of hydrogen bondrather than ionic. The amount of bound 3H-indirubin was directly proportional to the con-centration of indirubin. However, it increased abruptly when the concentration of indirubinreached 1.5×10-4 M.展开更多
Objective To test the immunogenicity of recombinant plasmid DNA containing human papillomavirus type 16 L1 (HPV16 L1) coding sequence of mice Methods The HPV16 L1 encoding sequence was generate...Objective To test the immunogenicity of recombinant plasmid DNA containing human papillomavirus type 16 L1 (HPV16 L1) coding sequence of mice Methods The HPV16 L1 encoding sequence was generated by polymerase chain reaction (PCR), and inserted into TA cloning vector PCR Ⅱ, then cloned in the eukaryotic expression vector pcDNA3 1 with CMV promoter The recombinant plasmid DNA pcDNA L1 was transferred into Cos 7 cells and used to immunize BALB/c mice via muscular injection The expression of HPV16 L1 in transferred cells was identified by immunospot and immunocytochemistry, which tested specific anti HPV16 L1 antibody in the serum of immunized mice Results Using the immunospot technique, we found L1 protein expression in pcDNA L1 transferred cells The immunocytochemistry studies demonstrated that the L1 protein was located in nuclei In immunized mice, specific anti HPV16 L1 antibodies could be detected by immunospot and immunocytochemistry 28 days after the first immunization and last at least 41 days Conclusions We constructed HPV16 L1 eukaryotic expressing plasmid whose DNA could induce immuno humoral response in mice This observation will be helpful in designing HPV16 prophylactic vaccine展开更多
Allele frequencies for 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) were obtained from 7,636 unrelated ...Allele frequencies for 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) were obtained from 7,636 unrelated individuals of Chinese Han population living in Qinghai and Chongqing, China. Totally 206 alleles were observed, with the corresponding allele frequencies ranging from 0.0001-0.4982. Chi-square test showed that all of the STR loci agreed with the Hardy-Weinberg equilibrium. We also compared our data with previously published population data of other ethnics or areas. The results are valuable for human identification and paternity testing in Chinese Han population.展开更多
This study evaluated the performance of the Wuxi AGCU ScienTech Incorporation(HuiShan,Wuxi,China)AGCU Expressmarker 16(EX 16)and 22(EX22)short tandem repeat(STR)amplification kits in reduced reaction volumes using dir...This study evaluated the performance of the Wuxi AGCU ScienTech Incorporation(HuiShan,Wuxi,China)AGCU Expressmarker 16(EX 16)and 22(EX22)short tandem repeat(STR)amplification kits in reduced reaction volumes using direct polymerase chain reaction(PCR)amplification workflows.The commercially available PowerPlex21(PP21)System(Promega,Wisconsin,USA),which follows similar direct workflows,was used as a reference.Anticoagulate blood applied to chemically impregnated FTATM Micro Cards(GE Healthcare UK Limited,Amersham Place,Little Chalfont,Buckinghamshire,HP79NA,UK)was used to represent a complex biological sample.Allelic concordance,first‑pass success rate,average peak heights,heterozygous peak height ratios(HPHRs),and intracolor and intercolor peak height balance were determined.In reduced volume PCR reactions,the performances of both the EX16 and EX22 STR amplification kits were comparable to that of the PP21 System.The level of performance was maintained at PCR reaction volumes,which are 40%of that recommended.The EX22 and PP21 System kits possess comparable overlapping genome coverage.This study evaluated the performance of the AGCU EX16 and EX22 STR amplification kits in reduced PCR reaction volumes using direct workflows in combination with whole blood applied to FTATM Micro Cards.Allelic concordance,first‑pass success rate,average peak heights,HPHRs,and intracolor and intercolor peak height balance were determined.A concordance analysis was completed that compared the performance of the EX16 and EX22 kits using human blood applied to FTA Micro Cards in combination with full,half,and reduced PCR reaction volumes.The PP21 System(Promega)was used as a reference kit.Where appropriate,the distributions of data were assessed using the Shapiro‑Wilk test.For normally‑distributed data,statistics were calculated using analysis of variance(ANOVA)and for nonparametric data the Wilcoxon/Kruskal‑Wallis test was used.Statistical significance was set at P<0.05.Confidence intervals for mean values were set at 95%.On using reduced volume PCR reactions in combination with dried blood spots applied to FTA sample collection cards,both the EX16 and EX22 kits were shown to generate STR profiles of sufficient quality to allow entry into National DNA databases.The performance of both EX16 and EX22 was comparable to that of the PP21 System.This study demonstrates the successful use of the Wuxi AGCU ScienTech Incorporation EX16 and EX22 kits in reduced PCR reaction volumes with complex biological samples applied to chemically impregnated FTA sample collection cards.展开更多
The EX 16+22Y system is a polymerase chain reaction(PCR)-based amplification kit that enables typing of 15 autosomal short tandem repeat(STR)loci(i.e„D3S1358,D13S317,D7S820,D16S539,TPOX,TH01,D2S133&CSF1PO,D19S433,...The EX 16+22Y system is a polymerase chain reaction(PCR)-based amplification kit that enables typing of 15 autosomal short tandem repeat(STR)loci(i.e„D3S1358,D13S317,D7S820,D16S539,TPOX,TH01,D2S133&CSF1PO,D19S433,vWA,D18S51,D21S11,D8S1179,D5S81&and FGA)and 22 widely used Y chromosome STR(Y-STR)loci(DYS391,DYS527a/b,DYS635,DYS458,DYS456,DYS385a/b,DYS43&DYS44&DYS437,DYS19,DYS576,DYS533,DYS393,DYS389I/n,DYS439,DYS392,Y_GATA_H4,DYS390,and DYS481)which contains 20 core Y-STR recommended by the Ministry of Public Security and amelogenin.This multiplex system was designed for the simultaneous analysis of amelogenin-Y allele mutation,single-source searches,kinship(including familial searching),mixture profiles,international data sharing,and other forensic applications.In this study,the multiplex system was validated for sensitivity of detection,species specificity,DNA mixtures,stability,sizing precision,stutter,reproducibility,and PCR-based conditions according to the Scientific Working Group on DNA Analysis Methods developmental validation guidelines and Chinese criteria for the human fluorescent STR multiplex PCR reagent.The results show that the EX16+22Y system is a robust and reliable amplification kit which can be used for human identification testing.展开更多
OBJECTIVE: Homeopathic nosodes have seldom been scientifically validated for their anticancer effects. This study was conducted to examine if a recently developed hepatitis C nosode has demonstrable anticancer potent...OBJECTIVE: Homeopathic nosodes have seldom been scientifically validated for their anticancer effects. This study was conducted to examine if a recently developed hepatitis C nosode has demonstrable anticancer potential in cancer cells in vitro.METHODS: Anticancer effects of Hepatitis C 30C(Hep C 30), if any, were initially tested on three cancer cell lines, HepG2(liver cancer), MCF-7(breast cancer) and A549(lung cancer) and one normal liver cell line WRL-68 cells and subsequently a more thorough study using further scientific protocols was undertaken on HepG2 cells(against WRL-68 cells as the normal control) as HepG2 cells showed better anticancer response than the other two. Three doses, one at 50% lethal dose(LD50) and the other two below LD50, were used on HepG2 cells subsequently. Protocols like apoptosis induction and its possible signaling mechanism were deployed using immunoblots of relevant signal proteins and confocal microscopy, with particular reference to telomerase and topoisomerase Ⅱ(Top Ⅱ) activities, two strong cancer biomarkers for their direct relationship with divisional activities of cells and DNAs. RESULTS: Hep C 30 induced apoptosis, caused distorted cell morphology typical of apoptotic cells, increased reactive oxygen species generation and produced increased DNA nicks. Further it enhanced pro-apototic signal proteins like Bax, cytochrome c and inhibited anti-apoptotic signal proteins, Bcl-2, cytochrome c and caspase-3, changed mitochondrial membrane potential and caused externalization of phosphatidylserine. The drug also decreased expression of two cancer biomarkers, Top Ⅱ and telomerase, consistent with its anticancer effect. CONCLUSION: Hep C 30 has demonstrable anticancer effects against liver cancer cells in vitro.展开更多
A consensus sequence, encoding a putative DNA polymerase type B derived from a Polinton transposon, was assembled from the sex determination region of Xiphophorus maculatus. This predicted protein, which is 1,158 aa i...A consensus sequence, encoding a putative DNA polymerase type B derived from a Polinton transposon, was assembled from the sex determination region of Xiphophorus maculatus. This predicted protein, which is 1,158 aa in length, contains a DNAA_pol_B_2 domain and a DTDS motif. The DNA polymerase type B gene has about 10 copies in the haploid X. maculatus genome with one Y-specific copy. Interestingly, it has specific copies on the W chromosome in the X. maculatus Usumacinta strain (sex determination with female het- erogamety), which represent new markers for this type of sex chromosome in platyfish. This marker with W- and Y-specific copies suggests relationship between different types of gonosomes and allows comparing male and female heterogameties in the platyfish. Further molecular analysis of the DNA polymerase type B gene in X. maculatus will shed new light on the evolution of sex chromosomes in platyfish.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.82030058).
文摘The field of forensic DNA typing,often referred to as“DNA fingerprinting,”has evolved and expanded considerably since its beginnings in the mid-1980s.Originally,forensic DNA typing was primarily used for individual identification and criminal investigations,but it has evolved into a versatile discipline with a wide range of applications.This article addresses the growing scope of forensic genetics,which includes advances in DNA sequencing technologies,mixture analysis,body fluid identification,phenotypic profiling,forensic genealogy,microbiological analysis,exploration of novel markers,and ethical and legal considerations.These developments have enabled the analysis of difficult samples and provided comprehensive insights into the origins of biological evidence.In an ever-evolving landscape,forensic genetics continues to shape the future of forensic science by providing new tools and techniques that help deliver justice in an increasingly complex world.
基金supported by the National Key Research and Development Program of China(2018YFA0900100)the Natural Science Foundation of Tianjin,China(19JCJQJC63300)Tianjin University。
文摘DNA molecules are green materials with great potential for high-density and long-term data storage.However,the current data-writing process of DNA data storage via DNA synthesis suffers from high costs and the production of hazards,limiting its practical applications.Here,we developed a DNA movable-type storage system that can utilize DNA fragments pre-produced by cell factories for data writing.In this system,these pre-generated DNA fragments,referred to herein as“DNA movable types,”are used as basic writing units in a repetitive way.The process of data writing is achieved by the rapid assembly of these DNA movable types,thereby avoiding the costly and environmentally hazardous process of de novo DNA synthesis.With this system,we successfully encoded 24 bytes of digital information in DNA and read it back accurately by means of high-throughput sequencing and decoding,thereby demonstrating the feasibility of this system.Through its repetitive usage and biological assembly of DNA movable-type fragments,this system exhibits excellent potential for writing cost reduction,opening up a novel route toward an economical and sustainable digital data-storage technology.
文摘Purpose: To alert the medical community that whole exome sequencing can find accessory gene changes in well-known syndromes that alter preventive health care and management. Meaning: A collagen type VI gene change adds muscle weakness, hypermobility, and dysautonomia concerns to usual management considerations for Down syndrome. Methods: Commercial whole exome sequencing combined with clinical interpretation of DNA sequence change added new considerations to patient management and parental counsel. Results: An 11-year-old child with the trisomy 21 form of Down syndrome who was evaluated for extraordinary joint laxity had a heterozygous collagen type VI aspartic to glutamic acid (COL6A3 c.6360 C>G p.Asp2120Glu) gene change found by whole exome sequencing. The DNA variant was qualified as having strong relevance to the enhanced hypermobility due to prior association of collagen 6 gene changes with myopathy. Conclusions: Dual diagnosis of Ehlers-Danlos syndrome was not assigned because the patient lacked criteria like bruising, unusual scars, or selected dysautonomia symptoms. The concept of a hypermobility spectrum offers advantages for management of its constituent conditions if clinically guided ascertainment and DNA diagnostics are employed.
基金ThisworkwasfinanciallysupportedbyagrantfromtheEducationCommissionofLiaoningProvince (No 990 2 2 1 0 69)
文摘s To investigate the DNA polymorphism of Sporothrix schenckii ( S schencki i ) and to find the relationship between DNA patterns and geographic areas and clinical manifestations Method The total DNA was extracted with hexadecyltrimethyl ammonium bromide Random A mplification of Polymorphic DNA (RAPD) assay was used to study DNA typing of 24 strains of S schenckii collected from different areas and isolated from di fferent clinical types Results Of seven random primers used, three primers (OPAA11, OPD18 and OPB07) gave good reactions, the sequences of which were 5' ACCCGACCTG 3', 5' GAGAGCCAAC 3', 5 ' GGTGAC^GCAG 3' respectively The RAPD patterns of the 24 isolates were not completely identical, showing certain degrees of hereditary variability Differ ent isolates showed a common conserved DNA band with the same primer Different clinical types showed different genotypes Conclusion RAPD analysis is useful in DNA typing of S schenckii , the DNA band type of which is related to geographic origin and Clinical manifestation
文摘The binding between indirubin and calf thymus DNA in vitro has been verified by meansof the isotope labelling method, spectrophotometric method and thermal denaturation meas-urements. The λmax 207 nm of indirubin shifted toward longer wave length with decrease ofabsorbance after the incubation of indirubin with DNA. The escalation of Tm value of DNAinduced by indirubin was about 2.4°C and it was reproducible. The binding force between themwas rather weak, as indirubin molecules were easily released during the precipitation withalcohol or the gel filtration. The binding was not affected by sodium chloride even at high con-centration but greatly decreased (to 20-30% of the control) in the presence of 8 M urea.These results showed that the binding between indirubin and DNA might be of hydrogen bondrather than ionic. The amount of bound 3H-indirubin was directly proportional to the con-centration of indirubin. However, it increased abruptly when the concentration of indirubinreached 1.5×10-4 M.
文摘Objective To test the immunogenicity of recombinant plasmid DNA containing human papillomavirus type 16 L1 (HPV16 L1) coding sequence of mice Methods The HPV16 L1 encoding sequence was generated by polymerase chain reaction (PCR), and inserted into TA cloning vector PCR Ⅱ, then cloned in the eukaryotic expression vector pcDNA3 1 with CMV promoter The recombinant plasmid DNA pcDNA L1 was transferred into Cos 7 cells and used to immunize BALB/c mice via muscular injection The expression of HPV16 L1 in transferred cells was identified by immunospot and immunocytochemistry, which tested specific anti HPV16 L1 antibody in the serum of immunized mice Results Using the immunospot technique, we found L1 protein expression in pcDNA L1 transferred cells The immunocytochemistry studies demonstrated that the L1 protein was located in nuclei In immunized mice, specific anti HPV16 L1 antibodies could be detected by immunospot and immunocytochemistry 28 days after the first immunization and last at least 41 days Conclusions We constructed HPV16 L1 eukaryotic expressing plasmid whose DNA could induce immuno humoral response in mice This observation will be helpful in designing HPV16 prophylactic vaccine
文摘Allele frequencies for 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) were obtained from 7,636 unrelated individuals of Chinese Han population living in Qinghai and Chongqing, China. Totally 206 alleles were observed, with the corresponding allele frequencies ranging from 0.0001-0.4982. Chi-square test showed that all of the STR loci agreed with the Hardy-Weinberg equilibrium. We also compared our data with previously published population data of other ethnics or areas. The results are valuable for human identification and paternity testing in Chinese Han population.
文摘This study evaluated the performance of the Wuxi AGCU ScienTech Incorporation(HuiShan,Wuxi,China)AGCU Expressmarker 16(EX 16)and 22(EX22)short tandem repeat(STR)amplification kits in reduced reaction volumes using direct polymerase chain reaction(PCR)amplification workflows.The commercially available PowerPlex21(PP21)System(Promega,Wisconsin,USA),which follows similar direct workflows,was used as a reference.Anticoagulate blood applied to chemically impregnated FTATM Micro Cards(GE Healthcare UK Limited,Amersham Place,Little Chalfont,Buckinghamshire,HP79NA,UK)was used to represent a complex biological sample.Allelic concordance,first‑pass success rate,average peak heights,heterozygous peak height ratios(HPHRs),and intracolor and intercolor peak height balance were determined.In reduced volume PCR reactions,the performances of both the EX16 and EX22 STR amplification kits were comparable to that of the PP21 System.The level of performance was maintained at PCR reaction volumes,which are 40%of that recommended.The EX22 and PP21 System kits possess comparable overlapping genome coverage.This study evaluated the performance of the AGCU EX16 and EX22 STR amplification kits in reduced PCR reaction volumes using direct workflows in combination with whole blood applied to FTATM Micro Cards.Allelic concordance,first‑pass success rate,average peak heights,HPHRs,and intracolor and intercolor peak height balance were determined.A concordance analysis was completed that compared the performance of the EX16 and EX22 kits using human blood applied to FTA Micro Cards in combination with full,half,and reduced PCR reaction volumes.The PP21 System(Promega)was used as a reference kit.Where appropriate,the distributions of data were assessed using the Shapiro‑Wilk test.For normally‑distributed data,statistics were calculated using analysis of variance(ANOVA)and for nonparametric data the Wilcoxon/Kruskal‑Wallis test was used.Statistical significance was set at P<0.05.Confidence intervals for mean values were set at 95%.On using reduced volume PCR reactions in combination with dried blood spots applied to FTA sample collection cards,both the EX16 and EX22 kits were shown to generate STR profiles of sufficient quality to allow entry into National DNA databases.The performance of both EX16 and EX22 was comparable to that of the PP21 System.This study demonstrates the successful use of the Wuxi AGCU ScienTech Incorporation EX16 and EX22 kits in reduced PCR reaction volumes with complex biological samples applied to chemically impregnated FTA sample collection cards.
文摘The EX 16+22Y system is a polymerase chain reaction(PCR)-based amplification kit that enables typing of 15 autosomal short tandem repeat(STR)loci(i.e„D3S1358,D13S317,D7S820,D16S539,TPOX,TH01,D2S133&CSF1PO,D19S433,vWA,D18S51,D21S11,D8S1179,D5S81&and FGA)and 22 widely used Y chromosome STR(Y-STR)loci(DYS391,DYS527a/b,DYS635,DYS458,DYS456,DYS385a/b,DYS43&DYS44&DYS437,DYS19,DYS576,DYS533,DYS393,DYS389I/n,DYS439,DYS392,Y_GATA_H4,DYS390,and DYS481)which contains 20 core Y-STR recommended by the Ministry of Public Security and amelogenin.This multiplex system was designed for the simultaneous analysis of amelogenin-Y allele mutation,single-source searches,kinship(including familial searching),mixture profiles,international data sharing,and other forensic applications.In this study,the multiplex system was validated for sensitivity of detection,species specificity,DNA mixtures,stability,sizing precision,stutter,reproducibility,and PCR-based conditions according to the Scientific Working Group on DNA Analysis Methods developmental validation guidelines and Chinese criteria for the human fluorescent STR multiplex PCR reagent.The results show that the EX16+22Y system is a robust and reliable amplification kit which can be used for human identification testing.
文摘OBJECTIVE: Homeopathic nosodes have seldom been scientifically validated for their anticancer effects. This study was conducted to examine if a recently developed hepatitis C nosode has demonstrable anticancer potential in cancer cells in vitro.METHODS: Anticancer effects of Hepatitis C 30C(Hep C 30), if any, were initially tested on three cancer cell lines, HepG2(liver cancer), MCF-7(breast cancer) and A549(lung cancer) and one normal liver cell line WRL-68 cells and subsequently a more thorough study using further scientific protocols was undertaken on HepG2 cells(against WRL-68 cells as the normal control) as HepG2 cells showed better anticancer response than the other two. Three doses, one at 50% lethal dose(LD50) and the other two below LD50, were used on HepG2 cells subsequently. Protocols like apoptosis induction and its possible signaling mechanism were deployed using immunoblots of relevant signal proteins and confocal microscopy, with particular reference to telomerase and topoisomerase Ⅱ(Top Ⅱ) activities, two strong cancer biomarkers for their direct relationship with divisional activities of cells and DNAs. RESULTS: Hep C 30 induced apoptosis, caused distorted cell morphology typical of apoptotic cells, increased reactive oxygen species generation and produced increased DNA nicks. Further it enhanced pro-apototic signal proteins like Bax, cytochrome c and inhibited anti-apoptotic signal proteins, Bcl-2, cytochrome c and caspase-3, changed mitochondrial membrane potential and caused externalization of phosphatidylserine. The drug also decreased expression of two cancer biomarkers, Top Ⅱ and telomerase, consistent with its anticancer effect. CONCLUSION: Hep C 30 has demonstrable anticancer effects against liver cancer cells in vitro.
基金supported by the grants from the Biofuture Programme of the German Federal Ministry for Education and Research (BMBF,to JNV),the French Research Agency (ANR to JNV)the Scientific Research Foundation for the Returned Overseas Chinese Scholars,the Ministry of Education of China (to QZ)
文摘A consensus sequence, encoding a putative DNA polymerase type B derived from a Polinton transposon, was assembled from the sex determination region of Xiphophorus maculatus. This predicted protein, which is 1,158 aa in length, contains a DNAA_pol_B_2 domain and a DTDS motif. The DNA polymerase type B gene has about 10 copies in the haploid X. maculatus genome with one Y-specific copy. Interestingly, it has specific copies on the W chromosome in the X. maculatus Usumacinta strain (sex determination with female het- erogamety), which represent new markers for this type of sex chromosome in platyfish. This marker with W- and Y-specific copies suggests relationship between different types of gonosomes and allows comparing male and female heterogameties in the platyfish. Further molecular analysis of the DNA polymerase type B gene in X. maculatus will shed new light on the evolution of sex chromosomes in platyfish.