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1.
Journal of Forensic Medicine ; (6): 267-279, 2022.
Article in English | WPRIM | ID: wpr-984120

ABSTRACT

In recent years, more and more forensic genetics laboratories have begun to apply massively parallel sequencing (MPS) technology, that is, next-generation sequencing (NGS) technology, to detect common forensic genetic markers, including short tandem repeat (STR), single nucleotide polymorphism (SNP), the control region or whole genome of mitochondrial DNA (mtDNA), as well as messenger RNA (mRNA), etc., for forensic practice, such as individual identification, kinship analysis, ancestry inference and body fluid identification. As the most widely used genetic marker in forensic genetics, STR is currently mainly detected by capillary electrophoresis (CE) platform. Compared with CE platform, MPS technology has the advantages of simultaneous detection of a large number of genetic markers, massively parallel detection of samples, the polymorphism of sequence detected by NGS makes STR have the advantages of higher resolution and system efficiency. However, MPS technology is expensive, there is no uniform standard so far, and there are problems such as how to integrate MPS-STR data with the existing CE-STR database. This review summarizes the current status of the application of MPS technology in the detection of STR genetic markers in forensic genetics, puts forward the main problems that need to be solved urgently, and prospects the application prospect of this technology in forensic genetics.


Subject(s)
DNA Fingerprinting/methods , Forensic Genetics/methods , Genetic Markers , High-Throughput Nucleotide Sequencing/methods , Microsatellite Repeats/genetics , Polymorphism, Single Nucleotide , Sequence Analysis, DNA , Technology
2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 188-195, 2021.
Article in Chinese | WPRIM | ID: wpr-906191

ABSTRACT

Objective:To evaluate the effect of Xuebijing injection on inflammatory indexes and immune function in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Method:PubMed,Cochrane Library,Embase,Wed of Science,CBM,CNKI,VIP and Wanfang Data Online Knowledge service platform were searched by computer,all of which were up to February 2020. After literature screening and quality evaluation by two researchers independently,relevant data were extracted and Meta-analysis was carried out by RevMan 5.3 software. Result:A total of 21 randomized controlled trials(RCT) were included,involving 1 618 patients. The results of Meta-analysis showed,Xuebijing injection adjuvant therapy was significantly better than the control group in total effective rate [relative risk(RR)=1.20,95% confidence interval(CI)(1.14,1.27),<italic>P</italic><0.000 01],C-reactive protein(CRP)[standard mean difference(SMD)=-1.58,95% CI(-1.98, -1.19),<italic>P</italic><0.000 01],total white blood cell (WBC)[mean difference(MD)=-1.44,95% CI(-1.84,-1.04),<italic>P</italic><0.000 01],procalcitonin(PCT)[SMD=-0.57,95% CI(-0.74,-0.41),<italic>P</italic><0.000 01],interleukin-6(IL-6)[SMD=-1.51,95% CI(-2.07,-0.96),<italic>P</italic><0.000 01],percentage of neutrophils(N%)[MD=-5.35,95% CI(-7.13,-3.58),<italic>P</italic><0.000 01],and tumor necrosis factor-<italic>α</italic>(TNF-<italic>α</italic>)[SMD=-1.52,95% CI(-2.23,-0.81),<italic>P</italic><0.000 1], and had positive effects in regulating cellular immune disorders. Conclusion:Xuebijing injection combined with routine treatment can improve the immune function of AECOPD patients,reduce the number of inflammatory markers,neutrophils and CD8<sup>+</sup> T cells, thus modulating the small airway microcirculation to promote inflammatory absorption and inhibit the progression of the disease, with high safety.

3.
Journal of Preventive Medicine ; (12): 988-993, 2021.
Article in Chinese | WPRIM | ID: wpr-905038

ABSTRACT

Objective@#To evaluate the impact of fine particulate matter ( PM2.5 ) on respiratory diseases and symptoms of community residents in Chun’an County, so as to provide the basis for air pollution treatment strategies. @*Methods@#Using the cluster random sampling method, the permanent residents in Qiandaohu Town of Chun’an County were investigated from 2017 to 2018. The demographic information, respiratory diseases and symptoms were collected by using Surveillance Plan for the Impact of Air Pollution ( Haze ) on health ( 2016 Edition ). The air pollutants and meteorological data were collected through Qiandaohu station of Hangzhou Ecology and Environment Monitoring Center and Hangzhou Meteorological Information Center. The effects of PM2.5 on respiratory diseases and symptoms of residents were analyzed with generalized estimating equation. @*Results@#Totally 1 181 people aged 6 months to 95 years were recruited, including 557 ( 47.16% ) males and 624 ( 52.84% ) females. Acute nasopharyngitis (common cold) and tracheitis/tonsillitis occurred most frequently, with 203 cases, accounting for 3.44%. The median of daily average concentration of PM2.5 was 24 μg/m3, with the standard exceeding rate of 2.80%. The results showed that PM2.5 increased the risk of acute nasopharyngitis ( common cold ), tracheitis/tonsillitis, cough, expectoration, runny nose, sore throat and nasal congestion ( lag 3 days, OR: 1.015-1.022, 95%CI: 1.001-1.037 ); the effect of PM2.5+PM10 ( OR: 1.020-1.040, 95%CI: 1.006-1.070 ) and PM2.5+O3 ( OR: 1.017-1.024, 95%CI: 1.005-1.035 ) was greater than that of PM2.5 alone on respiratory diseases and symptoms.@*Conclusion@#Atmospheric PM2.5 exposure in Chun’an County increases the risk of respiratory diseases and symptoms among community residents.

4.
Journal of Acupuncture and Tuina Science ; (6): 452-457, 2020.
Article in Chinese | WPRIM | ID: wpr-872437

ABSTRACT

Objective: To observe the clinical efficacy of heat-sensitive moxibustion plus Western medicine in treating patients with diabetic peripheral neuropathy (DPN). Methods: A total of 70 patients with DPN were divided into an observation group and a control group by sealed envelope method combined with the random number table method, with 35 cases in each group. The control group was treated with routine medicine, and the observation group was treated with heat-sensitive moxibustion on the basis of the treatment in the control group. After 2 courses of treatment, the scores of Toronto clinical scoring system (TCSS) and vibration perception threshold (VPT) in both groups were observed, and the clinical efficacy was compared. Results: During treatment, 3 cases dropped out in the control group and 4 cases in the observation group. After treatment, the total effective rate in the observation group was higher than that in the control group (P<0.05). The scores of TCSS and VPT in both groups decreased after treatment, and the intra-group comparison showed statistical significance (both P<0.05). The scores of TCSS and VPT in the observation group were lower than those in the control group, and the differences were statistically significant (both P<0.05). Conclusion: Heat-sensitive moxibustion plus Western medicine can improve the symptoms in patients with DPN, and has a better curative effect than the Western medicine alone.

5.
Journal of Forensic Medicine ; (6): 807-809, 2020.
Article in Chinese | WPRIM | ID: wpr-985179

ABSTRACT

Postmortem interval (PMI) estimation has always been an important and difficult issue in the field of forensic pathology. In recent years, research progress on the estimation of PMI using RNA specific variation patterns after death has been made by researchers at home and aboard. This paper summarizes the specific application methods of messenger RNA and non-coding RNA for PMI estimation based on the literatures and discusses the existing problems and development trends, in order to provide technical reference for related studies and estimation practice.


Subject(s)
Humans , Autopsy , Forensic Pathology , Postmortem Changes , RNA, Messenger/genetics , RNA, Untranslated , Time Factors
6.
Journal of Forensic Medicine ; (6): 387-392, 2019.
Article in English | WPRIM | ID: wpr-985022

ABSTRACT

Objective Quantitative analysis and comparison of the expression of ribonucleic acid (RNA) from frozen organs and formaldehyde-fixed and paraffin-embedded (FFPE) tissues. Methods Frozen specimens of human brain, myocardium and liver tissues as well as FFPE samples at different postmortem intervals were collected and mass concentration of RNA was extracted and detected. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technology was used to analyze the amplification efficiency and relative expression of each RNA marker. Results The mass concentration and integrity of RNA extracted from FFPE samples were relatively low compared with frozen specimens. The amplification efficiency of RNA markers was related with RNA species and the length of amplification products. Among them, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin (ACTB) with relatively long amplification products failed to achieve optimal amplification efficiency, whereas 5S ribosomal RNA (5S rRNA) achieved ideal amplification efficiency and showed quite stable expression across various tissues, therefore it was chosen as internal reference marker. The expression quantity of GAPDH and ACTB in frozen specimens with longer postmortem intervals and in FFPE samples with relatively long amplification products was decreased. The expressions of tissue-specific microRNAs (miRNAs), GAPDH and ACTB with relatively short amplification products had consistency in the same tissues and FFPE samples. Conclusion Through standardizing the RT-qPCR experiment, selecting the appropriate RNA marker and designing primers of appropriate product length, RNA expression levels of FFPE samples can be accurately quantified.


Subject(s)
Humans , DNA Primers , Formaldehyde , Gene Expression Profiling , MicroRNAs/analysis , Myocardium , Paraffin Embedding , RNA/analysis , Real-Time Polymerase Chain Reaction/standards
7.
Journal of Experimental Hematology ; (6): 1765-1771, 2018.
Article in Chinese | WPRIM | ID: wpr-774388

ABSTRACT

OBJECTIVE@#To investigate the effects of different stimultors (PHA, PMA and IL-2) and culture systems (PBMC and whole blood) on the proliferation of human peripheral blood lymphocyte subsets, so as to provide the experimental basis for selecting the appropriate system according to the experimental purposes.@*METHODS@#A total of 10 ml serum samples were collected from healthy volunteers (n=6). The 300 μl whole blood was directly used to detect lymphocyte subsets by flow cytometry. The 400 μl whole blood were inoculated respectively with 3 different stimulators at 37℃ and 5% CO2 for 60 h; Three different stimulators were also added to the PBMC which were isolated from 2 ml whole blood. Then the proliferation ability of lymphocyte subsets was analyzed by flow cytometry.@*RESULTS@#After the PBMC were stimulated with PHA, CD4CD8CD3 lymphocytes were the most subset; The proportion of CD3CD4 T lymphocytes and CD3CD19 B lymphocytes decreased after being stimulated by PMA (P<0.01, P<0.05); the lymphocyte subset ratio had no significant change after being stimulated by IL-2. After the whole blood system was stimulated with PHA, the CD4/CD8 T lymphoblasts were main subsets, the counts of B lymphocytes and NK cells were reduced; after being stimulated with PMA, the number of CD8CD3 T lymphoblast and CD4CD8T lymphocytes increased, the B/NK cells were not distinguished with the surface markers; after the whole blood system was stimulated with IL-2, the proportion of NK cells significantly increased (P<0.05).@*CONCLUSION@#Lymphocyte proliferation stimulated by PMA is the fastest, while the effect of IL-2 on the lymphocyte subset proportion stimulated by IL-2 is the minimal. After being stimulated by PHA the division cycles of lymphocyte are the most.


Subject(s)
Humans , Cell Proliferation , Flow Cytometry , Killer Cells, Natural , Lymphocyte Activation , Lymphocyte Count , Lymphocyte Subsets
8.
Journal of Regional Anatomy and Operative Surgery ; (6): 326-329, 2018.
Article in Chinese | WPRIM | ID: wpr-702272

ABSTRACT

Objective To explore the corresponding rules between neural process and frontal lobe gyri and provide anatomic basis for locating frontal lobe gyri by neural process. Methods 20 normal cadaver heads were transected into brain slices with thickness of 6 mm after dyeing frontal lobe gyri. Typical planes were observed,while the correspondence between neural process and frontal lobe gyri on coronal sec-tions were analyzed and summarized. Results There was 1 or 2 processes for almost frontal lobe gyri,with the direction of neural process in different coronal sections being unchanged and symmetric. Conclusions The corresponding rules between neural processes and frontal lobe gyri may be obtained and the frontal lobe gyri on coronal sections may be located through neural processes.

9.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 400-406, 2018.
Article in Chinese | WPRIM | ID: wpr-695678

ABSTRACT

Objective·To compare the quality of RNA extracted from fresh and formalin-fixed paraffin-embedded (FFPE) brain tissues and to explore the long non-coding RNA (IncRNA) expression level.Methods·FFPE samples stored under various conditions and paired frozen brain tissues were collected and total RNA qualities were then detected.Amplification efficiency (AE) and expression stability of each RNA marker were calculated and analyzed based on real-time quantitative PCR.After selecting reference biomarkers,normalized △ Ct values of candidate makers within different amplicon size were measured to assess the possibility of lncRNA quantification in FFPE tissues.Results·The purity of RNA extracted from FFPE was relatively high,but the RNA integrity was lower than fresh samples.All biomarkers were successfully amplified and amplification efficiencies of long-chain RNA markers were correlated with amplicon sizes,sample treatment and preservation conditions,namely temperature and storage time.5S,miR-9 and miR 125b achieved optimal AE and showed quite stable expression in all specimens,therefore they were chosen as control markers.Compared with fresh samples,the △ Ct values of only 2 lncRNA (HAR1F and MALAT1-L,whose amplicon size were both higher than 200 bp,respectively) increased in the FFPE samples kept in 4 ℃,while in FFPE tissues kept in room temperature,increments of the △ Ct values were significant for most target genes except for short amplicon markers (<60 bp),which showed consistently stable expression in all brain specimens.Conclusion·RNA integrity is affected by sample treatment and preservation conditions,but IncRNA expression levels in FFPE tissues can be accurately quantificated by using optimal amplicon sizes and considerable reference markers.

10.
Journal of Acupuncture and Tuina Science ; (6): 96-103, 2018.
Article in Chinese | WPRIM | ID: wpr-712656

ABSTRACT

Objective:To observe the therapeutic efficacy of acupuncture plus Tai Ji Quan (Tai Chi) in recovering the neurological function and treating depression state in post-stroke depression patients,together with a 12-month follow-up.Methods:A total of 105 eligible post-stroke depression patients were randomized into an acupuncture plus Tai Ji group (53 cases) and a control group (52 cases) based on their visiting sequence.The patients all received routine treatment and rehabilitation training for stroke.In addition,the control group was given oral administration of citalopram hydrobromide tablets,1 month as a course of treatment,for 3 courses in total.Meanwhile,the acupuncture plus Tai Ji group received acupuncture and practiced Tai Ji Quan,for 1 month and 12 months respectively.Before the intervention,after 1-month intervention and 12 months later,the National Institute of Health stroke scale (NIHSS),Barthel index (BI) and Hamilton depression rating scale (HAMD) were adopted for efficacy evaluation.Results:Prior to the intervention,there were no significant differences in HAMD,NIHSS and BI scores between the two groups (all P>0.05);after 1-month intervention,there were significant between-group differences in NIHSS,BI and HAMD scores (P<0.05 or P<0.01);the 12-month follow-up revealed significant between-group differences in NIHSS,BI and HAMD scores (all P<0.01).In the treatment of stroke,the total effective rate was 84.4% in the acupuncture plus Tai Ji group,significantly higher than 68.9% in the control group (P<0.05);in the treatment of depression,the total effective rate was 86.7% in the acupuncture plus Tai Ji group,significantly higher than 77.8% in the control group (P<0.05).Conclusion:Acupuncture plus Tai Ji Quan can produce a significant efficacy in improving the limb motor function and depression in post-stroke depression patients.

11.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 400-406, 2018.
Article in Chinese | WPRIM | ID: wpr-843726

ABSTRACT

Objective: To compare the quality of RNA extracted from fresh and formalin-fixed paraffin-embedded (FFPE) brain tissues and to explore the long non-coding RNA (lncRNA) expression level. Methods: FFPE samples stored under various conditions and paired frozen brain tissues were collected and total RNA qualities were then detected. Amplification efficiency (AE) and expression stability of each RNA marker were calculated and analyzed based on real-time quantitative PCR. After selecting reference biomarkers, normalized △ Ct values of candidate makers within different amplicon size were measured to assess the possibility of lncRNA quantification in FFPE tissues. Results: The purity of RNA extracted from FFPE was relatively high, but the RNA integrity was lower than fresh samples. All biomarkers were successfully amplified and amplification efficiencies of long-chain RNA markers were correlated with amplicon sizes, sample treatment and preservation conditions, namely temperature and storage time. 5S, miR-9 and miR-125b achieved optimal AE and showed quite stable expression in all specimens, therefore they were chosen as control markers. Compared with fresh samples, the △ Ct values of only 2 lncRNA (HAR1F and MALAT1-L, whose amplicon size were both higher than 200 bp, respectively) increased in the FFPE samples kept in 4 ℃, while in FFPE tissues kept in room temperature, increments of the △ Ct values were significant for most target genes except for short amplicon markers (<60 bp), which showed consistently stable expression in all brain specimens. Conclusion: RNA integrity is affected by sample treatment and preservation conditions, but lncRNA expression levels in FFPE tissues can be accurately quantificated by using optimal amplicon sizes and considerable reference markers.

12.
Pakistan Journal of Pharmaceutical Sciences. 2017; 30 (2): 363-367
in English | IMEMR | ID: emr-186495

ABSTRACT

A complex of low-molecular cationic peptides, extracted from human urine by a combination membrane ultrafiltration and weak cation exchange chromatography, was characterized in this study. It provides a simpler solution for the development of novel antimicrobial peptides from biological liquid waste

13.
Journal of Southern Medical University ; (12): 205-209, 2016.
Article in Chinese | WPRIM | ID: wpr-232483

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the correlation between a diverse of clinical factors and bone metastases of prostate cancer.</p><p><b>METHODS</b>The clinical data of 80 patients with prostate cancer were collected and analyzed. The correlations of age, alkaline phosphotase (ALP), prostate specific antigen (PSA), erythrocyte sedimentation rate (ESR), Gleason score, and expressions of androgen receptor (AR) and Ki-67 with bone metastases were analyzed by one-way ANOVA and Logistic regression analysis. The cutoff value, sensitivity and specificity of the independent correlation factors were calculated.</p><p><b>RESULTS</b>Forty-five of the 80 patients (56%) were found to have bone metastasis, who had significantly older age and higher levels of ALP, PSA, ESR, Gleason score, and expressions of AR and Ki-67 than those without bone metastasis (P<0.05). Logistic regression analysis identified PSA, Gleason score and AR expression as independent factors correlated with bone metastasis with OR (95% CI) of 1.005 (1.001, 1.009) (P=0.008), 5.356 (1.431, 20.039) (P=0.013), and 18.594 (2.460, 140.524) (P=0.005), respectively. The cutoff values of PSA, Gleason Score and AR were 67.1 ng/ml, 7.5, and 2.5, respectively; their sensitivities were 55.6%, 75.6%, and 84.0% for predicting bone metastasis with specificities of 97.1%, 82.9%, and 91.4%, respectively.</p><p><b>CONCLUSION</b>Of the factors analyzed, PSA, Gleason score and AR expression, but not age, ALP, PSA, ESR, or Ki-67 expression, are the predictive factors of bone metastasis of prostate cancer.</p>


Subject(s)
Humans , Male , Alkaline Phosphatase , Metabolism , Bone Neoplasms , Diagnosis , Neoplasm Grading , Predictive Value of Tests , Prostate-Specific Antigen , Blood , Prostatic Neoplasms , Pathology , Receptors, Androgen , Metabolism , Sensitivity and Specificity
14.
International Eye Science ; (12): 2166-2168, 2016.
Article in Chinese | WPRIM | ID: wpr-638069

ABSTRACT

AIM: To discuss the clinical efficacy of fresh amniotic membrane ( AM ) during the microscopic adjustable suture surgery in children's intercommunity strabismus, in order to guide clinical treatment. METHODS: With the clinical randomized control study (RCT), 60 (112 eyes) cases of patients in childhood who received microscopic strabismus surgery in our hospital were divided them into two different groups from Jan. 2010 to Oct. 2015. According to the application of AM on the basis of ophthalmology outpatient number, 30 cases (58 eyes) in group A were treated with rectus muscle recession surgery combined adjustable suture combined with AM. The other 30 cases (54 eyes) in group B were treated with rectus muscle recession surgery combined adjustable suture only. All patients in two groups were followed-up over 6mo after the strabismus surgery. RESULTS:Twenty-seven cases ( 48 eyes ) of all the strabismus patients must be adjusted after strabismus surgery, and the eye position adjustment rate was 42.9%. At 1mo after surgery, eye position of 18 cases (29 eyes) can be adjusted in all patients, and 44. 8% (16 cases, 26 eyes ) in group A with the average of adjustment lengths was 2. 56±0. 64mm, and 5. 6% ( 2 cases, 3 eyes ) in group B, with the average of adjustment lengths was 0. 52±0. 28mm, the differences of the adjustment rate and the average of adjustment amount were both high statistically significant (χ2 =22.477, P CONCLUSION: The application of fresh AM in the microscopic adjustable suture strabismus surgery is exactly effective in treatment of children's intercommunity strabismus. It can significantly extend the adjustment time and increase the adjustment amount, and it also can statistically improve the controllability and achievement ratio for children's strabismus surgery.

15.
Journal of Forensic Medicine ; (6): 245-249, 2016.
Article in Chinese | WPRIM | ID: wpr-984839

ABSTRACT

OBJECTIVES@#To explore the correlation between the expression levels of several RNA markers in human brain tissue and early postmortem interval (PMI).@*METHODS@#Twelve individuals with known PMI (range from 4.3 to 22.5 h) were selected and total RNA was extracted from brain tissue. Eight commonly used RNA markers were chosen including β-actin, GAPDH, RPS29, 18S rRNA, 5S rRNA, U6 snRNA, miRNA-9 and miRNA-125b, and the expression levels were detected in brain tissue by real-time fluorescent quantitative PCR. The internal reference markers with stable expression in early PMI were screened using geNorm software and the relationship between its expression level and some relevant factors such as age, gender and cause of death were analyzed. RNA markers normalized by internal reference were inserted into the mathematic model established by previous research for PMI estimation using R software. Model quality was judged by the error rate calculated with estimated PMI.@*RESULTS@#5S rRNA, miRNA-9 and miRNA-125b showed quite stable expression and their expression levels had no relation with age, gender and cause of death. The error rate of estimated PMI using β-actin was 24.6%, while GAPDH was 41.0%.@*CONCLUSIONS@#5S rRNA, miRNA-9 and miRNA-125b are suitable as internal reference markers of human brain tissue owing to their stable expression in early PMI. The expression level of β-actin correlates well with PMI, which can be used as an additional index for early PMI estimation.


Subject(s)
Humans , Actins/analysis , Autopsy , Brain/metabolism , MicroRNAs/analysis , Models, Theoretical , Postmortem Changes , RNA Stability , RNA, Ribosomal, 18S/analysis , RNA, Ribosomal, 5S/analysis , RNA, Small Nuclear/analysis , Real-Time Polymerase Chain Reaction , Software
16.
Journal of Forensic Medicine ; (6): 165-170, 2016.
Article in English | WPRIM | ID: wpr-984828

ABSTRACT

OBJECTIVES@#To explore the correlation between early postmortem interval (PMI) and eight RNA markers of rat's brain at different temperatures.@*METHODS@#Total 222 SD rats were randomly divided into control group (PMI=0 h) and four experimental groups. And the rats in the experimental groups were sacrificed by cervical dislocation and respectively kept at 5 ℃, 15 ℃, 25 ℃ and 35 ℃ in a controlled environment chamber. The RNA was extracted from brain tissues, which was taken at 9 time points from 1 h to 24 h postmortem. The expression levels of eight markers, β-actin, GAPDH, RPS29, 18S rRNA, 5S rRNA, U6 snRNA, miRNA-9 and miRNA-125b, were detected using real-time fluorescent quantitative PCR, respectively. Proper internal reference was selected by geNorm software. Regression analysis of normalized RNA markers was performed by SPSS software. Mathematical model for PMI estimation was established using R software. Another 6 SD rats with known PMI were used to verify the mathematical model.@*RESULTS@#5S rRNA, miR-9 and miR-125b were suitable as internal reference markers for their stable expression. Both β-actin and GAPDH had well time-dependent degradation patterns and degraded continually with prolongation of PMI in 24 h postmortem. The mathematical model of the variation of ΔCt values with PMI and temperature was set up by R software and the model could be used for PMI estimation. The average error rates of model validation using β-actin and GAPDH were 14.1% and 22.2%, respectively.@*CONCLUSIONS@#The expression levels of β-actin and GAPDH are well correlated with PMI and environmental temperature. The mathematical model established in present study can provide references for estimating early PMI under various temperature conditions.


Subject(s)
Animals , Rats , Actins/metabolism , Autopsy , Brain/pathology , Genetic Markers , MicroRNAs , Models, Theoretical , Postmortem Changes , RNA Stability , RNA, Small Nuclear , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Regression Analysis , Software , Temperature , Time Factors
17.
Journal of Experimental Hematology ; (6): 1791-1795, 2015.
Article in Chinese | WPRIM | ID: wpr-272520

ABSTRACT

CCAAT/enhancer binding protein alpha gene (CEBPA) is an important transcription factor in maintenance of differentiation of granulocyte series of hematopoietic system. It plays a key role in regulating cell proliferation and differentiation. CEBPA mutation easily occurs in M1 and M2 type of acute myeloid leukemia, about 5%-14% in adult acute myeloid leukemia and 7.9% in children with acute myeloid leukemia. At present, domestic CEBPA mutation research is far less than abroad. This review focuses on the structual characteristics and detection method of CEBPA, CEBPA clinical features, the effect of CEBPA mutation on the prognosis of patients and the choice of treatment.


Subject(s)
Humans , CCAAT-Enhancer-Binding Proteins , Leukemia, Myeloid, Acute , Mutation , Prognosis
18.
Journal of Forensic Medicine ; (6): 245-249, 2014.
Article in Chinese | WPRIM | ID: wpr-983910

ABSTRACT

OBJECTIVE@#To explore the correlation between postmortem interval (PMI) and five RNA markers of rat's skin--β-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S ribosomal RNA(18S rRNA), 5S ribosomal RNA (5S rRNA), and microRNA-203 (miR-203), at different temperatures.@*METHODS@#Eighteen SD rats were randomly divided into three environmental temperature groups: 4 °C, 15 °C and 35 °C, respectively. Skin samples were taken at 11 time points from 0 h to 120 h post-mortem. The total RNA was extracted from the skin samples and the five RNA levels were detected by real-time fluorescent quantitative PCR. Proper internal reference was selected by geNorm software. Regression analysis of the RNA markers was conducted by GraphPad software.@*RESULTS@#5S rRNA and miR-203 were most suitable internal references. A good linear relationship between PMI and RNA levels (β-actin and GAPDH) was observed in two groups (4 °C and 15 °C), whereas the S type curve relationship between the expression levels of the two markers (β-actin and GAPDH) and PMI was observed in the 35 °C group. The partial linear relationship between 18S rRNA and PMI was observed in the groups (15 °C and 35 °C).@*CONCLUSION@#Skin could be a suitable material for extracting RNA. The RNA expression levels of β-actin and GAPDH correlate well with PMI, and these RNA markers of skin tissue could be additional indice for the estimation of PMI.


Subject(s)
Animals , Rats , Actins , Autopsy , Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Postmortem Changes , RNA , RNA Stability , RNA, Ribosomal, 18S , Real-Time Polymerase Chain Reaction , Regression Analysis , Skin , Temperature
19.
Journal of Forensic Medicine ; (6): 7-12, 2014.
Article in Chinese | WPRIM | ID: wpr-983871

ABSTRACT

OBJECTIVE@#To observe the changes of relative expression of myocardial various RNAs in rats died of different causes and their relationship with PMI.@*METHODS@#The rat models were established in which the rats were sacrificed by broken neck, asphyxia, and hemorrhagic shock. Total RNAs were extracted from myocardium. The quantitative real time PCR was used to calculate threshold cycle values of RNAs including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-actin, inducible nitric oxide synthase (iNOS), hypoxia-inducible factor-1 (HIF-1), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and U6 small nuclear RNA (U6 snRNA) and to study the changes of the relative expressions of various indexes with PMI.@*RESULTS@#U6 snRNA with stable expression level could be used as appropriate internal control. In the early PMI, the relative expression of GAPDH, HIF-1, iNOS, TNF-alpha, and IL-6 more characteristically increased in groups of asphyxia and hemorrhagic shock than in group of broken neck, but the quantity of beta-actin decreased in all groups. In the late PMI, all the relative expressions significantly declined in correlation with the degradation of RNA.@*CONCLUSION@#The characteristic changes of each RNA expression can be used as references to estimate PMI in deaths by different causes.


Subject(s)
Animals , Rats , Actins , Cause of Death , Cytokines/metabolism , Disease Models, Animal , Enzymes/metabolism , Glyceraldehyde-3-Phosphate Dehydrogenases , Myocardium/metabolism , Nitric Oxide Synthase Type II , RNA/metabolism , RNA, Small Nuclear , Shock, Hemorrhagic , Tumor Necrosis Factor-alpha
20.
Journal of Experimental Hematology ; (6): 16-19, 2014.
Article in Chinese | WPRIM | ID: wpr-264958

ABSTRACT

This study was aimed to explore the clinical characteristics and therapeutic efficacy of normal karyotype AML patients with CEBPA mutations. Fifty-five de novo AML patients with normal karyotype were retrospectively analyzed with regard to frequency of CEBPA mutation, clinical characteristics and therapeutic response. The results showed that CEBPA mutation was detected in 20 patients (36.4%), among them 17 cases displayed double mutations, three cases were with single mutation. The clinical characteristics of patients with CEBPA mutation displayed as follows: 75% of AML patients with CEBPA mutation were AML-M1 and AML-M2, the hemoglobin level at newly diagnosis was higher and the platelet count at newly diagnosis time was lower than those of AML patients without CEBPA mutation [(98.30 ± 20.33) g/L vs (81.69 ± 23.74) g/L (P < 0.05); and (33.30 ± 38.27) ×10(9)/L vs (64.79 ± 61.60) ×10(9)/L (P < 0.05)]. The leukemic cells highly expressed CD7 and CD34. The therapeutic efficacy of 1 cycle for AML patients with CEBPA mutation was satisfactory (72.2%), was higher than that of patients without CEBPA mutation(68.6%), but there was no statistical significance (P > 0.05). It is concluded that AML with CEBPA mutation is more observed in AML-M1 and AML-M2, and accompanies by high level of hemoglobin and lower platelet count, expression of CD7 and CD34. Early-term therapeutic efficacy is satisfactory. The frequency of CEBPA mutation may be higher in Chinese patients with AML compared with that reported in Western world.


Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , CCAAT-Enhancer-Binding Proteins , Genetics , Karyotype , Karyotyping , Leukemia, Myeloid, Acute , Diagnosis , Genetics , Therapeutics , Mutation , Prognosis , Retrospective Studies , Treatment Outcome
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