3.DNA quantification of blood samples pre-treated with pyramidon.
Chuan-Hong ZHU ; Dao-Li ZHENG ; Rao-Zhi NI ; Hai-Sheng WANG ; Ping NING ; Hui FANG ; Yan LIU
Journal of Forensic Medicine 2014;30(3):188-190
OBJECTIVE:
To study DNA quantification and STR typing of samples pre-treated with pyramidon.
METHODS:
The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology.
RESULTS:
In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples.
CONCLUSION
Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology*
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Blood Stains
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DNA/isolation & purification*
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DNA Fingerprinting
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Forensic Medicine
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Humans
;
Polymerase Chain Reaction
;
Reproducibility of Results
;
Specimen Handling
4.The difference in distribution of HCV genotypes between patients infected with HCV by transfusion and non-transfusion routes.
Yao XIE ; Hui ZHAO ; Wei-ni OU ; Jie YAN ; Shao-cai DU ; Dao-zhen XU
Chinese Journal of Experimental and Clinical Virology 2004;18(3):247-250
OBJECTIVETo investigate the HCV genotypes distribution in northern and southern cities in China and the difference between patients infected with HCV by transfusion and non-transfusion routes.
METHODSThe HCV genotypes of the patients with chronic hepatitis C from 9 cities belonging to different regions were genotyped by the PCR products of 5 prime untranslated region NTR digested with restriction endonucleases, and the HCV genotypes distribution among different cities or between the patients infected with HCV through transfusion and other routes was analyzed.
RESULTSThe HCV genotypes of 214 in 219 cases were determined; 197 patients were infected with monogenotype HCV. The major epidemic genotypes of HCV isolates in China were 1b (76.64%) and 2a (18.22%), but 5.14% of patients were infected with HCV belonging to genotype 3b and this was the first report that there is genotype 4a in China. The HCV genotype distribution was not different in northern and southern areas, but was significantly different between patients infected with HCV through transfusion and non-transfusion routes (P=0.036). In patients infected trough transfusion, the rates of monogenotype HCV infection and genotype 1b were 93.88% and 76.87%, respectively, which were higher than those (86.57% and 58.21%) in the patients infected with HCV through non-transfusion routes. The rate of patient infected with mixed genotype HCV strains in non-transfusion group was 13.43%, which was higher than that (6.12%) of patients in transfusion group.
CONCLUSIONThe HCV genotype distribution in northern and southern regions were similar, but was significantly different between the patients infected through transfusion and other routes.
5' Untranslated Regions ; Adolescent ; Adult ; Aged ; China ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; isolation & purification ; Hepatitis C, Chronic ; etiology ; genetics ; transmission ; Humans ; Male ; Middle Aged ; Transfusion Reaction
5.Apoptosis and apoptosis-related genes in experimental autoimmune inner ear disease.
Liang CHAI ; Yang GAO ; Zhi-yan GU ; Dao-feng NI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2005;40(8):561-565
OBJECTIVETo investigate the protein and mRNA expression patterns of apoptosis-related genes, together with evidence of apoptosis, in relation to experimental autoimmune inner ear disease (AIED).
METHODSMale C57BL/6 mice at 4 weeks age (n = 80) were randomly assigned to one of the five group (n = 16). The inbred mice were given a single subcutaneous injection of diluted solution of pertussis and an emulsion containing equal parts of complete Freund adjuvant (CFA) and inner ear antigens (IEAg) extracted form guinea pig. The animals were sacrificed for inner ear examination at a defined time after the immunization (7, 14, 21 or 28 days). An autoimmune inner ear diseases model was established. Apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (d-UTP) nick end-laying (TUNEL) method. Using immunohistochemical techniques and reverse transcriptase polymerase chain reaction to clarify the profile of Fas, FasL, and bcl-2.
RESULTSUnder normal conditions, no TUNEL-positive cell was observed in the cochlea except for a few positive cells in the supporting cells of Corti's organ and macula sacculi. Inner ear antigens administration induced TUNEL-positive reactions in a wide variety of cells such as inner hair cells, supporting cells, stria vascularis and spiral ligament fibrocytes. No positive staining was evident in outer hair cells, spiral ganglion cells and Scarpa's ganglion cells during the whole period. Fas proteins were expressed in a wide range of cells in inner ear. The levels of Fas mRNA were no significant differences between normal and AIED mice. FasL and bcl-2 proteins could be detected in spiral ganglion cells and Scarpa's ganglion cells both in normal and AIED mice. FasL positive cells increased in number in inner ear of AIED mice. bcl-2 positive cells were not detectable in inner hair cells, stria vascularis and spiral ligament both in normal and AIED mice. The mRNA of three kinds of apoptosis-related genes was detectable in the normal and AIED mice. FasL mRNA was expressed at low levels in normal, being maximal at 14 d post inoculation and decreased gradually to steady levels by 2 weeks. The levels of bcl-2 mRNA increased significantly during the period of AIED.
CONCLUSIONApoptosis mediated by Fas/FasL signal system may play a role in the initiation and maintenance of AIED. bcl-2 has a crucial role in the regulation of the process of apoptosis in the inner ear of AIED mice.
Animals ; Apoptosis ; Autoimmune Diseases ; genetics ; immunology ; metabolism ; pathology ; Fas Ligand Protein ; genetics ; metabolism ; Guinea Pigs ; In Situ Nick-End Labeling ; Labyrinth Diseases ; genetics ; immunology ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; Spiral Ganglion ; metabolism ; fas Receptor ; genetics ; metabolism
6.Breast intraductal lesion resection under breast fiberoptic ductoscopy.
Wei WU ; Xiao-rong LI ; Kai-yan YANG ; Bo-ni DONG ; Dao-jin CHEN
Journal of Central South University(Medical Sciences) 2008;33(1):81-84
OBJECTIVE:
To explore the feasibility and to sum up the experience of breast intraductal neoplasm resection under breast fiberoptic ductoscopy (FDS).
METHODS:
FDS was performed on 548 patients with nipple discharge from Sep.2004 to Nov.2006. The clinical data of breast intraductal neoplasm found by FDS in patients who underwent tumor resection were analyzed, and the breast intraductal neoplasm image characteristics, diagnosis, operative type and postoperative pathological results were analyzed.
RESULTS:
Of the 548 patients with nipple discharge, intraductal neoplasm was found in 187 cases (34.1%), intraductal papilloma in 159 cases (29.0%), intraductal papillomatosis in 12 cases (2.2%), and breast carcinoma in 16 cases (2.9%). One hundred thirty-five patients were operated on in our hospital, of whom 91 were performed tumor resection or segmentectomy under the localization by FDS, and the other 44 were performed segmentectomy after breast duct infusion of methylene blue. The diagnostic rate under FDS in the FDS group (97.8%) was higher than that in the breast duct infusion methylene group (86.4%) (chi2=6.96, P=0.008).
CONCLUSION
FDS is not only an accurate diagnosis for breast intraductal lesion, but also an assistance to localize the breast intraductal neoplasm and to remove them in the operation.
Adult
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Aged
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Aged, 80 and over
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Breast Neoplasms
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diagnosis
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surgery
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Carcinoma, Ductal, Breast
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diagnosis
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surgery
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Endoscopy
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methods
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Female
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Fiber Optic Technology
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methods
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Humans
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Middle Aged
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Papilloma, Intraductal
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diagnosis
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surgery
7.Effects of telmisartan on endoplasmic reticulum stress induced cardiomyocyte apoptosis in abdominal aortic banded rats
Jia-Rong TANG ; Xiao-Ni YAN ; Chang-Qing ZHOU ; Li NI ; Jin-Jie HOU ; Dao-Wen WANG
Chinese Journal of Cardiology 2008;36(9):838-842
Objective To investigate the effects of telmisartan on endoplasm retiedum (ER) stress signal pathways and eardiomyocyte apoptosis in abdominal aortic banded rats.Methods Male SD rats were randomly divided into sham-operated group,abdominal aortic banding group(AAB)and AAB+telmisartan (5 mg·kg-1·d-1 per gavage,beginning at l week before AAB for 8 weeks,n=10 each).Ten weeks post AAB,hemodynamie measurements were performed,whole heart and left ventricular weights were obtained. Cardiomyocyte apoptosis was measured by TUNEL method.Myocardial GRP78 and CHOP protein expresssions were detected by Western blot and immunohistochemistry.Results The ratio of left ventricular weight to body weisht,the ratio of heart weight to body weisht,left ventrieular end diastolic pressure and the apoptosis index were significantly increased while left ventfieular end systolic pressure and±dp/dtmax were significantly decreased in AAB group than those in sham-operated group (all P<0.01),these changes could be significantly attenuated by telmisartan (all P<0.01).Moreover,myocardial GRP78 and CHOP expressions were significantly upregulated in AAB group than those in sham-operated group and telmisartan could significantly downregulate the increased GRfp78,CHOP expressions (all P<0.01).Conclusions Increased ER stress might be responsible for enhanced eardiomyocyte apoptosis in AAB rats.Telmisartan effectively attenuated the cardiomyocyte apoptosis and cardiac hypertrophy in AAB rats possibly through reducing ER stress.
8.Long-term efficacy of infant hepatitis B immunization program.
Jian GONG ; Rong-cheng LI ; Jin-ye YANG ; Yan-ping LI ; Xiu-rong CHEN ; Zhi-yi XU ; Chong-bai LIU ; Hui-lin CAO ; Kai ZHAO ; Dao-ming NI
Chinese Journal of Hepatology 2003;11(4):203-205
OBJECTIVETo evaluate the long-term efficacy of infant hepatitis B (HB) immunization program on preventing hepatitis B virus (HBV) infection, and to assess its impact on the incidence of HB in children.
METHODSSince 1986, the universal HB vaccination for newborn babies with standard, pediatric dose had been launched without serologic prescreening of pregnant women for HBsAg, in a high endemic county of Long-An. A hepatitis surveillance system was set up to evaluate the possible impact on the incidence of hepatitis B. To serologically evaluate the effectiveness of the program, a stratified random sampling of 1000 children in 1987 birth cohorts, who received plasma-derived HB vaccine, was recruited for long-term follow up at the age of 1 to 13 years. A cross-sectional seroepidemiological survey was conducted in the county in 1985, before the program, and in 2001, for 1551 children born in 1996-2000 who were administered yeast recombinant HB vaccine.
RESULTSDuring the 1 to 13 years after the program, the rates of HBsAg-positive were 0.7% to 2.9% with an average of 1.7% and the protective rates were 83.5% to 96.6%. HBV infection rates were 1.1% tp 5.1% with an average of 2.4% and the protective rates were 93.5% to 98.4%. For the population aged 1 to 4 years who were immunized with recombinant HB vaccine, HBsAg positive rates were 1.8% to 2.4% with an average of 2.0% and the protective rates were 78.4 to 85.2%. 14 years after the program, the cumulative incidence of acute hepatitis B in the children aged 1 to 14 years fell to 1.5 cases per 100,000 children, down 91.8% as compared with that in 1985 to 1987. However, the cumulative incidence of 14.4 cases per 100,000 population in unvaccinated children was not significantly different from that in the history controls. Acute hepatitis B children had not been reported, showing that the vaccination program was 100% protective in children.
CONCLUSIONThe universal infant HB vaccination program in a hyperendemic area has proved to be effective in controlling HBV infection and decreasing the incidence of acute hepatitis B in children. Booster dose is unnecessary in 13 years after the immunization. The protective efficacy of yeast recombinant HB vaccine is similar to that of plasma-derived HB vaccine.
Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Follow-Up Studies ; Hepatitis B ; epidemiology ; prevention & control ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B Vaccines ; immunology ; Humans ; Immunization Programs ; Incidence ; Infant ; Infant, Newborn ; Male ; Pregnancy ; Seroepidemiologic Studies ; Vaccination
9.Effects of drying temperature, growing area and plucking time on two saponins from leaves of Hedera helix
Li-Jun NI ; Deng LI ; Shao-Rong LUAN ; Li-Guo ZHANG ; Mei-Peng ZHU ; Zhi-Wen JIN ; Dao-Rong YAN ; Han-Tai JIN
Chinese Traditional Patent Medicine 2018;40(4):875-880
AIM To investigate the effects of drying temperature,growing area and plucking time on hederacoside C,α-hederin from leaves of Hedera helix L..METHODS Three batches of H.helix leaves plucked in different time from two growing areas were dried in a vacuum oven to the constant weight at 60 ℃,70 ℃,80 ℃,90 ℃ and 105 ℃,respectively.Two saponins in the processed leaves were determined by HPLC.The powders of the processed H.helix leaves of different batches were mixed with proper ratios,which were determined by least squares optimization method with constraints.RESULTS The content of hederacoside C in the processed H.helix leaves of the three batches increased while that of α-hederin decreased with increasing temperature.The relative error between measured value and desired contents of hederacoside C and α-hederin in the mixed H.helix leaves was less than 5.5%.CONCLUSION The effects of three factors on the content of two saponins in the H.helix leaves are in the order of drying temperature,growing area and plucking time.Mixing processed H.helix leaves of different quality statues reasonably can control the contents of two saponins in a certain range.
10.Clinical research of patients with congenital anosmia.
Jian-feng LIU ; Hui YOU ; Dao-feng NI ; Qia-hang ZHANG ; Xue-yan WU ; Na-ya WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2007;42(10):749-752
OBJECTIVETo report on a series of patients with congenital anosmia, and to discuss its classification and clinical characteristics.
METHODSEight patients with congenital anosmia were reviewed retrospectively. Four of eight cases were congenital anosmia with other abnormalities, including three cases with Kallmann's syndrome, one with hypoplasia of nasal cavity and nasal sinus. Four cases were isolated congenital anosmia. A thorough medical and chemosensory history, physical examination, nasal endoscopy, T&T olfactory testing, olfactory event-related potentials and sinonasal computed tomography scan were performed in all patients. Magnetic resonance image of olfactory pathway was available in seven cases, and olfactory biopsies were done in two cases.
RESULTSAll patients reported had never been able to smell anything. ENT physical examination and nasal endoscopy were normal, except one case with hypoplasia of nasal cavity. Subjective olfactory test indicated all of them were anosmia. No olfactory event-related potentials to maximum stimulus were obtained. Magnetic resonance imaging revealed the absence of olfactory bulbs and tracts in six cases, hypoplasia of bilateral olfactory bulbs and tracts in one case. Computed tomography scan indicated normal except hypoplasia of nasal cavity and sinus in one case. Three cases with Kallmann syndrome showed poor development of both primary and secondary sexual characteristics and had decreased serum luteinizing hormone, follicle-stimulating hormone, testosterone and estradiol.
CONCLUSIONSDiagnosis of congenital anosmia is established on chief complain, physical examination, nasal endoscopy, olfactory testing and olfactory imaging. Magnetic resonance imaging of olfactory pathway is indispensable.
Adolescent ; Adult ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Olfaction Disorders ; classification ; diagnosis ; Olfactory Pathways ; pathology ; Retrospective Studies ; Tomography, X-Ray Computed ; Young Adult