Inhibitory effects of apolipoprotein M on inflammatory factors induced by high glucose in human retinal vascular endothelial cells / 中华实验眼科杂志

Objective To observe the expressing changes of apolipoprotein M (ApoM),tumor necrosis factor-α(TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in human retinal vascular endothelial cells (HRECs) under the high glucose culture condition and investigate the inhibitory effects of ApoM overepression on the expressions of TNF-α and MCP-1.Methods HRECs were cultured in DMEM containing 10％ fetal bovine serum and 5.5 mmol/L D-glucose and assigned to 6 groups.The cells in the normal control group were cultured in above culture medium;the cells in the high glucose group were treated using the DMEM with 30 mmol/L D-glucose;ApoM was transfected into the cells using lentiviral vector in the ApoM transfected group;lentiviral vector without ApoM sequence was transfected in the empty vector group;the cells transfected by empty vector were cultured in high glucose culture medium in the empty vector+high glucose group;the cells in the ApoM transfection+high glucose group were treated by ApoM sequence transfection and high glucose incubation.The relative expression of ApoM,TNF-α and MCP-1 mRNA was detected using real-time quantitative PCR,and the relative expression of ApoM protein was evaluated using Western blot assay.Results Compared with the normal control group,the mRNA expression levels of ApoM,TNF-α and MCP-1 in the high glucose group were significantly increased (t=5.517,3.295,2.555;all P＜0.05).HRECs grew well after infected with lentivirus.The relative expression level of ApoM mRNA in the ApoM transfected group was 236.400±39.270,which was significantly higher than 1.000±0.153 in the empty vector group (t=5.995,P＜0.01).An enhanced protein band of ApoM was seen in the ApoM transfected group,and the protein band was absent in the empty vector group.The relative expression band in the ApoM transfected group was 1.000± 0.249 and 2.978 ± 0.285 in the cells cultured with normal culture medium or high glucose culture medium,respectively,with a significant difference between them (t =5.056,P＜0.01).The relative expressions of TNF-α and MCP-1 in the mRNA levels were significantly different among the empty vector group,empty vector+high glucose group,ApoM transfected group and ApoM transfection + high glucose group (F =5.966,P =0.026;F =14.410,P =0.002).Compared with the empty vector+high glucose group,the relative expressions of TNF-α and MCP-1 mRNA were considerably reduced in the ApoM transfection+high glucose group (P=0.017,0.004).Conclusions High glucose environment up-regulates the expression of ApoM,MCP-1 and TNF-α in HRECs.Overexpression of ApoM inhibits the up-regulation of MCP-1 and TNF-α expression induced by high glucose.

Application of Coronary Artery Disease-Reporting and Data System in risk factor analysis of patients with coronary artery disease / 吉林大学学报(医学版)

Objective:To investigate the application of Coronary Artery Disease-Reporting and Data System (CAD-RADS) in the diagnosis of coronary atherosclerotic heart disease(CAD) and its risk factors,and to clarify the effective strength of different risk factors in the diagnosis of CAD by using CAD-RADS.Methods:All the data of 266 patients,who were initially suspected with CAD and underwent CT angiography,were collected and diagnosed by using CAD-RADS and were divided into CAD group(n=174) and non-CAD group(n=69).The informations of age,gender,hypertension,hyperlipemia,diabetes,smoking,serum uric acid (UA) levels,and plasma fibrinogen (FIB) levels of the patients in two groups were analyzed;single factor analysis and multivariate Logstic regression analysis were used to analyze the risk factors of CAD diagnosed by CAD-RADS.Results:Compared with non-CAD group,the ratios of male,hypertension,diabetes,smoking of the patients in CAD group were increased (P<0.05),and the age and the level of UA of the patients in CAD group were also increased (P<0.05).The Logistic regression analysis results showed that age and diabetes were the independent risk factors for CAD diagnosed by CAD-RADS.Conclusion:There are many independent risk factors for CAD diagnosed by CAD-RADS,and age and diabetes are the most correlated risk factors for CAD.

The recent research advance in sphingosine 1-phosphate and its receptors affecting atherosclerosis / 天津医药

Sphingosine 1-phosphate (S1P) is one of the crucial signal molecules, which can regulate many biological functions inside and outside cells. It plays an important role in regulating numerous physiological and pathological processes after being combined with S1P receptors (S1PRs). S1P/S1PRs signaling pathways have become a hot spot in the current research on endothelial inflammation and atherosclerosis. This review described the current development of the role of S 1P and its receptors in atherosclerosis.

Comparison of diagnostic values between magnetic susceptibility weighted imaging and routine MRI in patients with cerebral amyloid angiopathy-related hemorrhage / 吉林大学学报(医学版)

Objective:To analyze the imaging features of magnetic susceptibility-weighted imaging (SWI) in the patients with cerebral amyloid angiopathy-related hemorrhage (CAAH),and to clarify the diagnostic value of SWI for CAAH.Methods:A total of 68 patients presumptively diagnosed as CAAH were collected,and their imaging data of routine MRI and SWI were collected and analyzed.The detection rates of hemorrhage focus of the CAAH patients were compared between two kinds of imaging examination.The consistency of detection of CAAH by routine MRI and SWI was analyzed.The imaging features of SWI and the risk of focus hemorrhage in the patients with CAAH were analyzed by multivariate Logistic regression analysis.Results:Sixty-one patients were confirmed as CAAH by pathologic diagnosis,and 53 patients were confirmed as CAAH by routine MRI;the detection rate was 86.89％;59 cases of hemorrhage focus were confirmed by SWI and the detection rate was 96.72％.The number of lesions detected by SWI was more than that of routine MRI (P＜0.05).The consistency of detection of CAAH by routine MRI and SWI was poor,and the value of Kappa was 0.3666.The patchy high signal and multiple clear edge low signal area were the relative imaging features of CAAH with SWI in the patients with CAAH analyzed by multivariate Logistic regression analysis (OR=3.895,P=0.025;OR=3.124,P=0.029).Conclusion:SWI can efficiently detect the hemorrhage focus in the patients with CAAH and the diagnostic value is better than routine MRI.

Comparison of diagnostic values between magnetic susceptibility weighted imaging and routine MRI in patients with cerebral amyloid angiopathy-related hemorrhage / 吉林大学学报(医学版)

Objective:To analyze the imaging features of magnetic susceptibility-weighted imaging (SWI) in the patients with cerebral amyloid angiopathy-related hemorrhage (CAAH),and to clarify the diagnostic value of SWI for CAAH.Methods:A total of 68 patients presumptively diagnosed as CAAH were collected,and their imaging data of routine MRI and SWI were collected and analyzed.The detection rates of hemorrhage focus of the CAAH patients were compared between two kinds of imaging examination.The consistency of detection of CAAH by routine MRI and SWI was analyzed.The imaging features of SWI and the risk of focus hemorrhage in the patients with CAAH were analyzed by multivariate Logistic regression analysis.Results:Sixty-one patients were confirmed as CAAH by pathologic diagnosis,and 53 patients were confirmed as CAAH by routine MRI;the detection rate was 86.89％;59 cases of hemorrhage focus were confirmed by SWI and the detection rate was 96.72％.The number of lesions detected by SWI was more than that of routine MRI (P＜0.05).The consistency of detection of CAAH by routine MRI and SWI was poor,and the value of Kappa was 0.3666.The patchy high signal and multiple clear edge low signal area were the relative imaging features of CAAH with SWI in the patients with CAAH analyzed by multivariate Logistic regression analysis (OR=3.895,P=0.025;OR=3.124,P=0.029).Conclusion:SWI can efficiently detect the hemorrhage focus in the patients with CAAH and the diagnostic value is better than routine MRI.

Identification and application of sphingosine-1-phosphate receptor of human umbilical vein endothelial cells / 天津医药

Objective To identify the expression of sphingosine-1-phosphate receptor (S1PR) subtypes, C-myc and His tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells (HUVEC), CRL-1730 for studying the function of apolipoprotein M (ApoM)-S1P axis. Methods Two kinds of cells (EA. hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was em?ployed to investigate the expressions of coagulation factorⅧ(FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Re?sults (1) Two kinds of cells both expressed FⅧand ApoM. FⅧpresented scattered particle distribution in CRL-1730, while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of two kinds of cells. (2) S1PR1-3 can be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression. (3) Two kinds of cells both expressed C-myc and His tag proteins in cytoplasm. Conclusion Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, C-myc and His tag proteins. It is not suitable for choosing C-myc and/or His tag–conjugated recombinant ApoM to study the fuction of ApoM-S1P axis with these two kinds of cells.

Establishing a method for detection of human vitamin D receptor using dual real-time fluorescence quantitative PCR / 天津医药

Objective To establish a dual real-time fluorescence quantitative polymerase chain reaction (dual real-time PCR) assay to detect human vitamin D receptor (VDR) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Methods GAPDH gene was used as the internal control. The specific primers and TaqMan probes were designed by Primer Premier 5.0 software, which were applied to detect the VDR/GAPDH mRNA levels. The obtained PCR products were puri-fied to construct the VDR/GAPDH recombinant plasmid, which was taken as the standard to analyze the sensitivity and re-peatability of the method. Results The amplification products were confirmed as the specific fragment of VDR/GAPDH by DNA sequencing instrument. The results showed that the sensitivity, linear range, the determinate coefficient, the amplifica-tion efficiency, the intra-assay and inter-assay coefficient of variation were 40 copies/μL, 4.00 × 101-4.00 × 105 copies/μL, 0.998, 96.10%, 0.09%-1.21%, 0.17%-0.51%for VDR, and 40 copies/μL, 4.00 × 101-4.00 × 105 copies/μL, 0.999, 85.15%, 0.35%-0.88%, 0.51%-2.46% for GAPDH, respectively. Conclusion These results demonstrate that the dual real-time PCR assay with high sensitivity and specificity can detect the relative expressions of human VDR by single reaction tube, which can effectively shorten the time and reduce the experimental error.

Expression of hsa-miR-10 b in three cervical cancer cell lines and its target genes / 基础医学与临床

Objective To analyze the expression of hsa-miR-10b in three cervical cancer cell lines , and to find the target genes of hsa-miR-10b.Methods PCR was applied to measure expression levels of hsa-miR-10b in C-33A, HeLa and CaSki .The relative studies on hsa-miR-10b were retrieved from PubMed .The sequence and genome char-acteristics of hsa-miR-10b were analyzed on line by miRbase and NCBI .The target genes of hsa-miR-10b were searched by TargetScan , PicTar and miRanda , and then demonstrated by Gene Ontology and Pathway Enrichment analysis.Results Compared with C-33A, the expression of hsa-miR-10b significantly reduced in HeLa and Caski (P<0.01).Current studies showed that hsa-miR-10b was related with multiple tumorigenesis .hsa-miR-10b was lo-cated in human chromosome 2q31.1 and highly conserved among different species .The target genes were enriched in the biological processes of transcription , gene expression regulation , cell proliferation and etc .(P<0.05).Pathway analysis showed that these target genes were responsible for biochemical erents mediated by to the signaling pathways of cancer, cell cycle, ARF and etc.(P<0.05).Conclusions hsa-miR-10b may have extensive functions , and closely related with the occurrence and development in cervical cancer .Prediction of target genes provides a theoreti-cal basis for the further study .

Development of a duplex fluorescence RT-PCR assay for identifying SRBⅠgene knockout mice / 天津医药

Objective To develop a duplex fluorescence RT-PCR assay for detection of scavenger receptor class B, typeⅠ(SRBⅠ) knockout mice. Methods Primers and probes were designed according to knockout region of SRBⅠgene and related substituted sequence. DNA samples were extracted from tails of mice and performed amplification using real-time PCR. SRBⅠgenotypes of mice were analyzed according to amplification curves of FAM and CY5 channels. Finally, the sensitivity of the method was detected and the accuracy was verified by the direct sequencing. Results The homozygous SRBⅠwild genotype showed an amplification curve only in FAM channel. When the homozygous SRBⅠknockout genotype was present, the typical S amplification curve appeared only in the CY5 channel. Heterozygous genotype showed two typical S amplification curves in both FAM and CY5 channels, respectively. The results showed that the sensitivity reached 4×101 copies/μL, and there was complete concordance between this method and direct DNA sequencing. Conclusion The new method is simple, rapid and accurate, which is suitable for genotyping SRBⅠknockout mice.

Association between a new SNP in proximal promoter region of apolipoprotein M and susceptibility of coronary artery diseases in Han Chinese population / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the association between genetic polymorphisms of proximal promoter region of apolipoprotein M (apoM) gene and susceptibility of coronary artery diseases (CAD) in Han Chinese population.</p><p><b>METHODS</b>Two pairs of primers were designed according to the sequence (GenBank accession nos. EU030444.1) and the PCR products of apoM proximal promoter region were directly sequenced. Two hundred and six patients [165 males, mean age (61.9 ± 9.2) years old] diagnosed with CAD according to the results of angiography (a lesion was classed as being significant when stenosis was more than 50%) were enrolled in the present study, 209 age- and gender-matched patients[157 males, mean age (60.4 ± 9.1) years old] without CAD according to the results of angiography were selected as the control group. The allelic frequencies and genotype distributions of polymorphism in CAD and non-CAD patients were analyzed. Furthermore the wide-type and mutant promoter region of apoM were cloned into the luciferase expression vector pGL3, respectively. Luciferase reporter assay was used to detect the activity of apoM promoter.</p><p><b>RESULTS</b>A new deletion mutation -724delC in apoM promoter was found. The frequency of Del C allele was 8.0% in CAD patients and only 4.1% in the non-CAD controls (OR = 2.054, 95%CI 1.125-3.749, P = 0.017). The mean TC level was lower in groups with wide-type homozygotes compared to the mutant allele carriers [ (6.04 ± 0.90) mmol/L vs. (4.95 ± 1.00)mmol/L, P < 0.01]. -724delC mutant showed obvious decreased luciferase activities (1.13 ± 0.25 vs. 2.11 ± 0.15, P = 0.009).</p><p><b>CONCLUSION</b>It is reasonable to speculate that -724delC could affect the activity of the apoM promoter and downregulate apoM expressions, therefore, influence the susceptibility of CAD in this patient cohort.</p>

Changes of serum apolipoprotein M level in women during normal and gestational diabetes mellitus / 中华内分泌代谢杂志

Objective To investigate the changes of serum apolipoprotein M (ApoM) level in cord blood and pregnant women during different gestational periods and to explore the relationship with the occurance and development of gestational diabetes mellitus (GDM).Methods Three hundred and eight pregnant women performing regular prenatal examination were registered in Changzhou Women and Children Health Hospital from June,2011 to April,2013,including 99 normal pregnant women during the second trimester,122 during the third trimester,and 87 pregnant women with GDM.At the same time,100 samples of cord blood of normal neonates were collected.The levels of serum ApoM and other lipids were measured.Results (1) Serum ApoM levels in pregnant women during the second trimester,during the third trimester,and in the cord blood of normal neonate were (0.036-± 0.013) g/L,(0.023 ± 0.008) g/L,and (0.010 ± 0.004) g/L,respectively.(2) The serum ApoM level of GDM group was lower than that of control [(0.029 ± 0.010 vs 0.036 ± 0.013) g/L,P＜0.05].Multivariate non-conditional logistic regression analysis showed that there was statistically significant difference in ApoM levels between GDM and control groups after adjusting for potential confounders (Z =-3.62,P＜0.05).Conclusions The level of serum ApoM in pregnant women was increased at first and then decreased.The serum ApoM level in the neonatal cord blood was significantly lower than that in pregnant women.The level of serum ApoM was lowered with raised blood glucose level in women with GDM,suggesting that ApoM might play a role in the pathological processes of GDM.

Establishment and Application of Hyperinsulinemic-Euglycemic Clamp Technique in Conscious Rats / 天津医药

Objective To establish the hyperinsulinemic-euglycemic clamp(HEC) technique in conscious rats, and to explore the effect of acute infusion of lipid on glucose infusion rate (GIR) in rats. Methods Ten SD rats were random-ly divided into two groups, 5 rats for each group. The right jugular vein and left carotid artery were catheterized and under-went a HEC with infusion of lipid (intralipid group) for 6 hours, and with continuing infusion of 5%glucose (control group). The plasma levels of free fatty acid(FFA) and GIR were measured by HEC method. Results The level of FFA concentration increased by 17.6-fold, and GIR was reduced by 27%in the intralipid group compared to those of control group (P＜0.001). Conclusion The rat model of HEC has been successfully established by intravenous intralipid infusion, which can be con-firmed by HEC technique.

Association of serum apolipoprotein A5 with leptin and insulin in obese children / 中华内分泌代谢杂志

Totally 79 obese children and 64 children with normal body weight were included in the present study.Serum apolipoprotein A5 (ApoA5) and leptin levels were determined by ELISA and fasting insulin by RIA.The clinical data including height,body weight,waist circumference,blood pressure,blood lipid,blood glucose,etc,were collected.Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated.The results showed that compared with normal weight children,both serum leptin and insulin levels were significantly raised in obese children [19.15 (13.01 ～ 25.08) ng/ml vs 3.29 (1.45 ～ 6.02) ng/ml and 15.44 (12.05 ～ 20.26) μg/L vs 10.12 (8.60 ～ 12.60) μg/L,both P＜0.01],while ApoA5 level was significantly lowered [134.5 (105.9 ～ 172.7) ng/ml vs 2005.9(164.3 ～ 265.3) ng/ml,P＜0.01].Serum ApoA5 was negatively correlated with serum leptin and insulin (both P＜0.01).

Estrogen increases apolipoprotein M through estrogen receptor / 中华内分泌代谢杂志

Objective To explore the effects of estrogen on apolipoprotein M ( apoM ).Methods ApoM mRNA was assayed in HepG2 cells by RT-PCR after incubation of estrogen with or without estrogen receptor antagonist at different concentrations and durations.SD female rats were divided into five groups:OVX group,Sham group,OVX+ EB group,normal group and normal + EB group.From a week of being operated,the rats were injected subcutaneously estradiol beuzoate or vehicle.After 12-hrs fasting,serum levels of triglycerides (TG),LDL-cholesterol,HDL-cholesterol,total cholesterol ( TC ) at months 1,2 and 3 after operation were measured.The expression of apoM in rats was detected by using real time RT-PCR and Western blot.Results Estrogen increased mRNA levels of apoM and apoAI in the HepG2 cells with a dose- and time-dependent manner,which could be abolished by addition of estrogen receptor antagonist.Serum apoM,TG,TC,HDL and LDL levels were significantly increased in the ovariectomized or normal rats which received estrogen treatment than those in OVX or normal group rats at month 1 after treatments.Conclusions Estrogen upregulates apoM expression via its receptor.

Expression and significance of human zinc finger 23 gene in hepatocellular carcinoma / 中华检验医学杂志

Objective To detect the expression level of human zinc finger 23 (ZNF23) in hepatocellular carcinoma tissue samples and HepG2 cell lines and investigate the relationship between hZNF23 expression and clinicopathological characteristics of HCC and cell apoptosis. Methods The expression levels of hZNF23 and GAPDH mRNA in 37 cases of HCC were measured by real-time RT-PCR. The association between the expression of hZNF23 and the clinicopathological characteristics of HCC was analyzed. Cultured HepG2 cells were divided into 4 groups ( control group, 1.25 μg/ml , 2.5 μg/ml and 5 μg/ml cisplatin)or 6 groups( control group, 1.25 μg/ml, 2.5 μg/ml, 5 μg/ml, 10 μg/ml and 20 μg/ml cisplatin). MTT method was employed to evaluate cell proliferation. Annexin V-FITC assay was used to assess percentage of apoptotic HepG2 cells. The expression levels of hZNF23 and GAPDH mRNA of HepG2 cells after apoptosis induced by cisplatin with a series of concentrations were measured by real-time RT-PCR.Results The median ( quartile1, quartile 3) expression levels of hZNF23 mRNA in 37 HCC tissue samples and adjacent tissue samples were 8.84 (3.59-15.05), 22.20 ( 13.85-42.90 ), respectively. There was significant difference ( U = 259.5, P ＜ 0.01 ). The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in cancer tissue samples with Edmondson stage Ⅰ + Ⅱ [12.80(4.80-19.50)] was much higher than those in stage Ⅲ + Ⅳ [5.01 ( 2.88-11.68 ), U = 99.00, P ＜ 0.05] The median ( quartile1,quartile 3 ) expression levels of hZNF23 mRNA in patients with and without hepatic cirrhosis were 9.92(3.80-15.25) , 3.21 (2.78-3.60), respectively. The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in HBV infection and non-infection patients was 9.09(3.72-15.25 ), 2.48 (1.79-12.10),respectively. There was no significant difference between groups with and without hepatic cirrhosis and between HBV infection and non-infection groups( U = 16. 00 and 24.00, P ＞0.05 ). MTT assay indicated that cisplatin significantly inhibited HepG2 cells proliferation in a dose-dependent manner. Annexin V-FITC/PI assay showed that HepG2 cells apoptosis rates were (0.9 ± 0.2 ) %, ( 4. 2 ± 0.3 ) %, ( 9.8 ± 4. 3 ) %,(23.0 ± 6.0)%, respectively. Cisplatin significantly induced HepG2 cells apoptosis in a dose-dependent manner( F = 27.89, P ＜ 0.01 ). The expression levels of hZNF23 mRNA in cisplatin groups [( 10.39 ±3.08) × 10-5, (24.10 ± 2.09) × 10-5, (6.90 ± 2.24) × 10-4] were significantly lower than that of the controlgroup[(94.80±1.80) ×10-5, F=6.027, P＜0.01]. Conclusions The expression level hZNF23 mRNA is related to Edmondson stage of HCC. The apoptosis effect of cisplatin on HepG2 cells may be associated with the upregulation of hZNF23.

Effect of gamma knife surgery on treatment of craniopharyngiomas / 中国基层医药

Objective To evaluate the adaption,effectiveness,dose,complication of the treatment for cranio pharygiomas with gamma knife surgery.Methods Gamma knife surgery was performed in 41 patients.Patients with mixed solid and cystic tumors were treated with stereotaxic aspiration while six cases were treated with divergence surgery prior to gamma knife therapy.The central dose ranges from 17.1 ～40 Gy(29.6Gy in average).The patients were treated at 30%～50% equal dose curve with 6 ～14Gy of tumormargin dose(9.5Gy in average).The exposure dose of the optic nerve and optic tract is less than 10Gy.Results Of twenty-nine patients who were followed up from 6 to 100 months,sixteen had disappeared or decreased tumor,six had unchanged,two was performed craniotomy one year or three years after gamma knife surgery,and the remaining five were dead one year to three years.The tumor control rate was 75.9%(22/29).Conclusion The treatment of stereotaxic radiation with single and high dose is sensitive to most of the solid craniopharyngioma,and the treatment of stereotaxic resection combined with gamma knife surgery may be feasible for the recurrent mixed solid and cystic craniopharyngioma.

A new TNM staging system inclusive of intraoperitoneal free cancer cells in gastric cancer patients / 中华普通外科杂志

Objective To evaluate a new TNM staging system inclusive of intraperitoneal free cancer cells in terms of postoperative survival of patients with gastric cancer. Methods Free cancer cells (FCC) in the peritoneal washes of gastric cancer patients were estimated by measuring CEA mRNA levels using real-time RT-PCR. After 5-year follow-up, we get the cut-off value of CEA mRNA level by using MedCalc software to analyze the ROC curve. When CEA levels are more than the cut-off value, it may considered as FCC(+), and then using FCC(+) as distant metastasis (MI) to make a new TNM staging and analyze patients life-span. Results (1) Under the ROC curve analysis, when the cut-off value of CEA mHNA level was at 31.21 copies/ml, the Youden's index is the highest. (2) When FCC (+) considered as M1 to make a new TNM staging, the 5-year survival rate showed as below: Ⅰ-Ⅱ, P=0. 134; stage Ⅱ-Ⅲ P=0.004 and Ⅲ-Ⅳ P=0.022,repecetively. Conclusion (1) The best cut-off value of CEA mRNA levels for FCC in peritoneal washes is 31.2 copies/ml. (2) Our study demonstrated that application of FCC(+) in the TNM staging may have a better estimation of prognosis of patients suffering from advanced gastric cancer.

Relationship between DNA of Rat Kidney and Post by Comet Assay / 中山大学学报(医学科学版)

[Objective] To study the correlation between kidney cell DNA degradation and postmortem interval within the span of 6-48 hour after the subject rats′ death.[Methods] To select 18 healthy mature female SD rats and equally divide them into 6 groups.To execute the rats with cervical spine articulation and put the rats under the incubator temperature of 25.1℃ (the average temperature of the 5 previous Decembers in Guangzhou prefecture).Sample kidney tissue from the rat separately 0 hour,6 hours,12 hours,24 hours,36 hours,48 hours,and 60 hours after the rats′ execution to prepare monoplast suspension,which is committed to comet assay.The comet images were captured by fluorescence CCD.Kinetic Comet 4.0 software was used to analyze images.Relevant data were collected by kinetic Comet 4.0 software and were subjected to Kruskal-Wallis test.[Results] Within the postmortem interval of 6-48 h,the number of SD rat kidney cell DNA fragments increased as the postmortem interval lengthens.So did the comet tail length.The Oliver tail moment and tail DNA of comet also showed sign of increase in positive proportion to the postmortem interval (their values corresponding to 60-hour-postmortem-interval were not obtainable.Kruskal-Wallis test indicated:the discrepancies of TL among the 6 groups were all significant (P ＜ 0.01).The difference of TM between 6 h group and 12 h group was not significant (P ＞ 0.05).The difference of TM between 24 h and 36 h was significant (P ＜ 0.05).The difference of TDNA among 6 h,12 h,and 36 h groups were not significant (P ＞ 0.05).The difference of TDNA between 36 h and 48 h was significant (P ＜ 0.05).[Conclusion] Degradation of nuclear DNA of the rat kidney cells increases as the postmortem interval lengthens and comet assay may provide important empirical evidence for determining the postmortem interval.

A postmortem semi-quantitative study on time-dependent changes of DNA content in chondrocytes in rat's costal cartilage / 中国法医学杂志

Objective To study the relationship between the DNA content of chondrocytes in the costal cartilage and postmortem interval in putrefactive rat cadavers.Methods Nuclear DNA wag visualized by modified Feulgen's staining method.DNA content of ehondrocytes in the costal cartilage was semi-quantita tively determined by a computerized image analysis system in rats within 35d postmortem.Results Staining intensity of the nuclei was gradually reduced within from 1d to 28d postmortem.The nuclej could not be detected at 35d.The DNA content of chondrocytes decreased time-dependently within 28 days after death as determined semi-quantitatively,which revealed a linear relationship between DNA content and postmortem interval.Conclusion DNA content of chondrocytes in the costal cartilage reduces time-dependently with the extension of postmortem interval.

A novel base-quenched probe technique for detecting single-nucleotide polymorphisms / 中华检验医学杂志

ncing analysis validated that all four-type base-quenched probes could provide unbiased genotyping results ( Kappa =1, P=0.00), although. Conclusion This method is simple, economic and suitable for large-scale genotyping studies.