Objective To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice,construct DXS6804 allelic ladder by molecular clonning and apply them in a population study on the Pumi population in Yunnan,China.Methods PCR was used to produce several different allelic fragments of the locus.After cloning the PCR products,the recombinant plasmids were sequenced.Then we denominated them and used them as template for re-amplification to generate the locus standard ladder.Results The sequencing results confirmed that the size and the construction of the inserts were correct.The genetic polymorphisms of this locus in Yunnan Pumi population of China were studied.Two off-ladder alleles of DXS6804 locus were found.Conclusion This method is of high value for forensic DNA typing to construct standard ladders.DXS6804 is robust for genetic research and forensic application.

The genesis of primary hepatocellular carcinoma(PHC)is a complicated biology process involve in multiple factors.It has been paid more attention to the genetic susceptibility in the pathogenesis and development of PHC.As the third generation of molecular genetic marker,single nucleotide polymorphism(SNP)becomes a hot topic in the research field of PHC.This article summarized the research evolution of the relationship between the susceptible gene SNP of PHC and PHC susceptibility.

Objective:To detect the clinical features of vulvar and vaginal intraepithelial neoplasias(VIN and VAIN,respectively).Methods:Total 148 women were performed vulvar or vaginal coloposcopy-directed biopsy pathology tests,from Sep.2004 to Dec.2007.Results:Among 148 women,vulvar or vaginal histologic results were vulvar cancer for 1,VIN or VAIN 2,3 for 23,VIN or VAIN for 16,condyloma for 61,vulvitis and vaginitis for 47.Eighty-five percent(33/39) women with VIN or VAIN 2,3 were more than 30 years old.Compared to women with cervical intraepithelial neoplasia(CIN),women with VIN or VAIN were older.The rate of high-risk HPV DNA in women with vulvar or vaginal lesions was 84%(84/100).VAIN occurred mainly in the upper vagina(90%,69/75).VIN or VAIN often accompanied or followed CIN or cervical cancer(79%,31/39),and VIN or VAIN 2,3 often accompanied or followed CIN 2,3 or cervical cancer(70%,16/23).Conclusion:Our data suggest that women with high-risk HPV infection are at risk of developing VIN or VAIN 2,3.The vulva and vagina should be carefully inspected by colposcopic examination at the time of colposcopy for any abnormal findings.

OBJECTIVETo observe the effects of electroacupuncture (EA) at "Dazhui" (GV 14) on Wnt-β-catenin signal pathway in annulus fibrosus cells in intervertebral disc in rats with cervical spondylosis.

METHODSForty SD rats were randomized into a control group, a model group, an EA group and a medication group, 10 rats in each one. Rats in the control group were treated with sham operation, only incision on local skin; rats in the remaining groups were made into cervical spondylosis models. After model establishment, rats in the control group and model group received fixed treatment under identical condition; rats in the EA group were treated with EA at "Dazhui" (GV 14), 30 min per treatment; rats in the medication group were treated with intragastric administration of meloxicam tablets. Treatments were both given once a day, and 14 days were taken as one session; there was an interval of 2 days between two sessions, and totally two sessions were given. After the treatments, immunohistochemistry was applied to measure the expression of Wnt, glycogen synthase kinase-3β (GSK-3β) and Axin in annulus fibrosus cells; western blot was used to test the expression of P-β-catenin.

RESULTSIn the control group, there were more positive cells of Wnt, GSK-3β and Axin, which were intensively distributed, deeply colored, and strongly positive; In the model group, there were less positive cells of Wnt, GSK-3β and Axin, which were sparsely distributed and weakly positive. The expression of Wnt, GSK-3β, Axin and P-β-catenin in the model group was less than that in the control group (all P < 0.05); expression of Wnt, GSK-3β, Axin and P-β-catenin in the EA group and medication group was higher than that in the model group (all P < 0.05); expression of Wnt, GSK-3β, Axin and P-β-catenin was not significantly different between EA group and medication group (all P > 0.05).

CONCLUSIONEA could delay the degeneration of intervertebral disc, which may be related to EA inhibiting signal pathway of Wnt-β-catenin.

Acupuncture Points ; Animals ; Electroacupuncture ; Female ; Fibrosis ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Humans ; Intervertebral Disc ; metabolism ; pathology ; Male ; Rats ; Spondylosis ; genetics ; metabolism ; pathology ; therapy ; Wnt Signaling Pathway ; beta Catenin ; genetics ; metabolism

Objective To study the chemical constituents from the exocarp of Juglans mandshurica.Methods The compounds were isolated by Diaion HP-20,Sephadex LH-20,MCI CHP-20,and silica gel column chromatography.Their structures were identified by physicochemical properties and spectroscopic analysis.Results Twelve compounds were elucidated as pinostrobin (Ⅰ),quercetin (Ⅱ),quercetin-3-O-?-D-glucoside (Ⅲ),chrysophanol (Ⅳ),gallic acid (Ⅴ),4-hydroxyl-cinnamic acid methyl ester (Ⅵ),vanillin (Ⅶ),caffeic acid (Ⅷ),4-hydroxylcinnamic acid (Ⅸ),?-sitosterol (Ⅹ),daucosterol (Ⅺ),and 5-hydroxyl-1,4-naphthoquinone (ⅩⅡ).Conclusion The compounds Ⅰ,Ⅳ,and Ⅵ—Ⅸ are isolated from J.mandshurica for the first time.

AIM:To study the effect of dexamethasone(Dex)on the expression of aquaporin-1(AQP-1)in cultured rat pleural mesothelial cells and to offer experimental support for the investigation of the mechanisms of pleural fluid treatment.METHODS:Rat pleural mesothelial cells were cultured in vitro.The expression of AQP-1 was detected by immunocytochemistry and reverse transcription-PCR(RT-PCR)after cells were identified,then the cells were treated with Dex for 24 hours at concentrations of 10-8,10-7,10-6,10-5 and 10-4mmol/L,and for 6 h,12 h,24 h,36 h,48 h,72 h at concentration of 10-4 mmol/L.Western blotting was used to analyze the expression of AQP-1 in cultured control and Dex-treated rat pleural mesothelial cells,and image analysis system to analyze the expression of AQP-1.RESULTS:Aquaporin-1 was expressed in cultured rat pleural mesothelial cells.The protein of AQP-1 expressed in rat pleural mesothelial cells was 755.04?19.81,843.72?19.41,862.96?26.53,694.80?32.00,938.08?13.32 in those treated with Dex at concentrations of 10-8,10-7,10-6,10-5 and 10-4 mmol/L,respectively,the levels was 2.02,2.26,2.31,1.86,2.52 fold higher than that in control group(372.90?16.46,P

A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components.

Object To assess the genetic diversity among ten natural populations in Ophiopogon japonicus (L f ) Ker Gawl and O. bodinieri Levl (Liliaceae) in Sichuan Province Methods Forty random decamer primers were screened for Random Amplified Polymophic DNA (RAPD) fragments Results Based on cluster analysis of 515 DNA bands amplified by 11 primers, a DNA molecular dendrogram was established, and the relationship of the populations and seeds between O japonicus and O bodinieri was set up Conclusion 1 Distinct genetic differences and extensive genetic diversity are presented among the samples The genetic differences are related to morphological differences, but not to geographic regions; 2 Quite genetic differences are presented between O japonicus and O bodinieri, it suggests that they have farther relationships

BackgroundIt has been well-known that integrin linked kinase(ILK) plays an important role in the pathogenesis of neorascularization.But there are few researches to elucidate the relationship between ILK and retinal nevascularization. Objective This study was to explore the expression and significance of ILK on retinal neovascularization and new vessels regression in the oxygen-induced retinopathy(OIR) mouse model. Methods One hundred and twenty-eight 7-day-old C57BL/6J mice were randomly divided into the normal group and model group.The mice were rose in(75±2)％O2 environment with mother mice together for 5 days and then in normal environment for 5 days to establish the OIR models.The mice in normal group were rose in the normal environment for 21 days.The 17-day-old mice were sacrificed and retinal sections were prepared for histopathological examination and the numbers of the cellular nuclei of vascular endotheliumbreaking retinalinternal limiting membranewere calculated.Retinal sections and sheeting were prepared in 12,14,17 and 21 -day-old mice to examine the formation and regression of new blood vessel using ADP histochemistry staining,and then immunochemistry,real-time PCR and Western-blotting were used to detect the expressions of ILK and its mRNA in retina. ResultsThe numbers of the cellular nuclei of vascular endothelium breaking retinal internal limiting membrane were (45.64 ± 12.17 )in OIR group,and those of the normal group were( 0.35±0.14 )with a significant difference between two groups (t =22.85,P＜0.05).Retinal new blood vessel appeared in 14-day mice,and peaked in 17-day mice and then regression in 21-day mice.ILK protein was expressed mainly in retinal ganglion cell layer,inner nuclear layer,inner plexiform layer and photoreceptor layer.Real-time PCR showed that ILK mRNA expressing levels in retina in model group were( 1.00±0.22),(1.85±0.17),(1.58±0.43) and(1.53±0.36) respectively in 12-,14-,17- and 21-day mice.Westernblotting determined that the A value of the ILK/β-actin was increased in 12-,14-,17-day mice and decreased in 21-day mice,and the A values were significantly higher in model group than the control group in various aged mice ( t =2.97,P＜0.05 ;t =11.88,P＜0.01 ;t =16.84,P＜0.01 ;t =13.00,P＜0.01 ). ConclusionsThese results indicate a space-time corresponding relation between the expression of ILK and retinal neovascularization.The obvious positive expression of ILK may be highly correlated with retinal neovascularization.