Objective To evaluate the feasibility of cell free fetal DNA(cffDNA)-based noninvasive prenatal diagnosis,we developed a precise technique for fetal sex determining region of Y chromosome(SRY)gene detection using size-fractionated cell-free DNA in maternal plasma.Methods Peripheral blood samples were collected form 117 pregnant women.cffDNA was extracted based on a column absorbent method and isolated by agarose gel electrophoresis.A dulex-polymerase chain reaction(PCR)was used to detected SRY gene and glycerol-dehyde-phosphate dehydrogenase(GAPDH)gene.Results Both SRY and GAPDH gene were detected in 86 cffDNA samples from women bearing male fetuses.And only GAPDH gene was detected in 71 cffDNA samples from women bearing female fetuses.These results had a coincidence whit those of villus or amniotic fluid samples.The specificity and sensitivity reached 100%(117/117)and 100%(66/66),respectively.Conclusion By agarose gel electrophoresis,re-extratedand and dulex PCR,size-fractionated cell-free fetal DNA in maternal plasma can be selective enriched and used to noninvasive prenatal diagnosis of sex-linked disorders and single gene disorders.

Mutation (substitution, deletion, insertion, etc.) in nucleotide acid causes the maximal sequence lengths of exact match (MALE) between paralogous members from a duplicate event to become shorter during evolution. In this work, MALE changes between members of 26 gene families from four representative species (Arabidopsis thaliana, Oryza sativa, Mus musculus and Homo sapiens) were investigated. Comparative study of paralogous' MALE and amino acid substitution rate (d(A)<0.5) indicated that a close relationship existed between them. The results suggested that MALE could be a sound evolutionary scale for the divergent time for paralogous genes during their early evolution. A reference table between MALE and divergent time for the four species was set up, which would be useful widely, for large-scale genome alignment and comparison. As an example, detection of large-scale duplication events of rice genome based on the table was illustrated.
Amino Acid Sequence ; Amino Acid Substitution ; Animals ; Arabidopsis ; genetics ; Chromosome Mapping ; methods ; Conserved Sequence ; genetics ; Evolution, Molecular ; Humans ; Mice ; Molecular Sequence Data ; Oryza ; genetics ; Proteome ; genetics ; metabolism ; Sequence Alignment ; methods ; Sequence Analysis, Protein ; methods ; Sequence Homology, Amino Acid ; Species Specificity

An ancient genome duplication (PPP1) that predates divergence of the cereals has recently been recognized. We report here another potentially older large-scale duplication (PPP2) event that predates monocot-dicot divergence in the genome of rice (Oryza sativa L.), as inferred from the age distribution of pairs of duplicate genes based on recent genome data for rice. Our results suggest that paleopolyploidy was widespread and played an important role in the evolution of rice.
Biological Evolution ; Chromosome Mapping ; methods ; Evolution, Molecular ; Genetic Variation ; genetics ; Genome, Plant ; Oryza ; genetics ; Polyploidy

Objective To evaluate the clinical value of ultrasonography guided percutaneous core needle biopsy in pancreatic lesions. Methods Thirty-four patients with 36 pancreatic lesions in Shanghai First People′s Hospital Afifliated to Shanghai Jiao Tong University from February 2012 to November 2013 underwent conventional ultrasound-guided percutaneous core needle biopsy using automatic gun and 18-gauge biopsy needles. The site, size, internal and surrounding vascularity, the sampling number of the lesions, and whether the specimens′ quality was satisfied were recorded. Then specimens were sent for pathological examination, and all above observations were compared with the ifnal diagnosis. Results The number of lesions with 2, 3 and 4 samplings was 32, 2 and 2, respectively. The average number of sampling was 2.2 (mean, 2.17;standard deviation, 0.51) and the acquisition rate of satisifed specimens was 89%(32/36). The pathological results of biopsy were malignant in 31 of 36 lesions including 27 cases of ductal adenocarcinoma, 2 cases of lymphoma, 1 case of small cell neuroendocrine carcinoma and 1 case of uterine leiomyosarcoma metastasis. The other 5 lesions were non-malignant including 3 cases of benign lesion, 1 cases of atypical hyperplasia and 1 cases of granulation tissue. The 36 lesions were ifnally diagnosed as 34 cases of pancreatic malignancy, 2 cases of non-malignant neoplasm. The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of ultrasonography guided percutaneous core needle biopsy in pancreatic lesions were 91%(31/34), 100%(2/2), 92%(33/36), 100%(31/31) and 40%(2/5), respectively. Youden index was 0.91. Two patients had mild upper abdominal pain and 1 patient had transient elevated serum amylase. No pancreatitis, pancreatic fistula, peritonitis, bleeding or dispersion of malignant cells along the penetrating channel or other serious complications occurred. Conclusion Ultrasonography guided percutaneous core needle biopsy is a simple, rapid, safe and effective diagnostic method in pancreatic lesions with high clinical value.

0.1),and there were significant differences in the other groups[F(H)=33.57,F(I)=133.6,F(HI)=69.75 Pa

Objective To investigate the effect of Ulinastatin on the delivery of cytokines in patients with septic shock.Methods It was a prospective and controlled clinical study.Seventy-eight patients with septic shock were randomly divided into control group and treatment group and thirty-nine in every group.Patients in treatment group received Ulinastatin 200 000 units intravenous everyday for 3 days,while those in control group received equal volume of normal saline as placebo.At different time points (at 24 th,48 th,72 th hour after start of treatment),the levels of tumor necrosis factor-alpha (TNF-?),interleukin-1 (IL-1),interleukin-6 (IL-6 ),interleukin-8 (IL-8) and superoxide dismutase (SOD) in serum were assayed.Results In comparison with control group,the levels of TNF-?,IL-1,IL-6,IL- 8 of treatment group decreased markedly (P＜0.05,P＜0.01) at different time points,whereas the level of SOD was higher markedly (P＜0.05,P＜0.01) at various time points.Conclusion Ulinastatin has protective effect on patients with septic shock through decreasing the levels of TNF-?,IL-1,IL-6,IL-8 and increasing in the level of SOD.

BACKGROUND:Sepsis has become the greatest threat to in-patients, with a mortality of over 25％.The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsis. This dysfunction occurs after surgery, but the magnitude of change does not differentiate patients with sepsis from those without sepsis. Increased intestinal permeability before surgery is of no value in predicating sepsis. The present study aimed to observe the changes of intestinal mucosal immunologic barrier in rat models of sepsis induced by cecal ligation and puncture. METHODS:Sixty Sprague-Dawley rats were randomly divided into a sepsis group (n=45) and a control group (n=15). The rats in the sepsis group were subjected to cecal ligation and puncture (CLP), whereas the rats in the control group underwent a sham operation. The ileac mucosa and segments were harvested 3, 6 and 12 hours after CLP, and blood samples were collected. Pathological changes, protein levels of defensin-5 (RD-5) and trefoil factor-3 (TFF3) mRNA, and lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected. RESULTS:The intestinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of lamina propria, hemorrhage and ulceration in the sepsis group. The expression of TFF3 mRNA and level of RD-5 protein were decreased and the apoptosis of mucosal lymphocyte increased (P<0.05) in the sepsis group compared with the control group. Significant differences were observed in RD-5 and TFF3 mRNA 3 hours after CLP and they were progressively increased 6 and 12 hours after CLP in the sepsis group compared with the control group (P<0.05, RD-5 F=11.76, TFF3 F=16.86 and apoptosis F=122.52). In addition, the gut-origin bacterial DNA detected in plasma was positive in the sepsis group. CONCLUSION:The immunological function of the intestinal mucosa was impaired in septic rats and further deteriorated in the course of sepsis.

Objective To study the level of serum lipids and body-fat content of high-fat diet induced pbesity rats(DIO).explore the relationship between intracellular caleium and ventrieufar arrhythmia.Methods Sixty male Sprague-Dawley(SD)rats were divided into control group(15)and experiment group(45),high-fat diet was administrated for 12 weeks to established obesity model,15 rats were selected into obesity group according to their body weight gain.The standard 2-lead electrocardiograph was used to detect the incidence and scores of arrhythmia induced by barium chloride(BaCl2,0.1 mg/kg)for 1 hour on every 8 rats from different groups respectively.Body-fat content.the level of serum total cholesterol(TC),triglyceride(TG),low density lipopmtein cholesterol (LDL-C) and high density lipopmtein cholesterol (HDL-C)were measured.The epididymal(EP),retroperitoneal (RE) and mesenteric(ME)white adipose pads was measured to obtain the body fat content.Single ventricular myrocytes of rats were isolated by enzymatic dissociation.The confocal laser scanning microscope was used to record basic intraceUular calcium level([Ca2+]i).Results The body-fat content in obesity group[(7.71±0.74)%] was significantly higher than control group[(4.69±0.37)%](t＝3.650,P＜0.05).The level of serum TC,TGand LDL-C were significantly higher(t＝3.801,2.778,3.536.P＜0.05) in obesity group[(1.26±0.04),(0.58±0.10),(0.51±0.04)mmol/L]than those in control group[(0.92±0.08),(0.29+0.03),(0.31±0.04) mmol/L].The level of serum HDL-C wa8 decreased gradually from control group[(0.53±0.05)mmol/L] toobesity group[(0.52±0.02)mmol/L],but there waft no significant difference between them(t＝0.186,P＞0.05).The incidence of arrhythmia induced by BaCl2(0.1 mg/kg)in obesity group was significantly higher than controlgroup(X2＝5.333,P＜0.05),and the scores of arrhythmla was increased in obesity group(2.5±0.6)too.The fluorescence intensity standing for[Ca2+]i was increased significantly(t＝2.409,P＜0.05)from obesity group(247.96±20.03)to control group (174.25±23.13).Conclusion As the free cytosolic calcium begin to accumulate,the arrhythmia morbidity is increased in obesity rats.

OBJECTIVETo explore a new noninvasive method for Hb Bart' s hydrops fetus by using PCR amplification efficiency discrimination between cell-free fetal DNA (cffDNA) and cell-free maternal DNA in maternal plasma.

METHODSCffDNA samples from pregnant women bearing possible Hb Bart's hydrops fetus were collected. Fluorescent PCR and capillary electrophoresis (CE) were performed. Hb Bart's hydrops fetus was conclusively identified by different peak area ratio of products.

RESULTSThe peak area ratio of 30 cffDNA samples from Hb Bart' s hydrops fetus was much less than 1. However, the ratio of cffDNA sample from hydrops fetus due to other reasons was approximately equal to 1.

CONCLUSIONBy using cffDNA fluorescent PCR and CE, a prenatal screening method for Hb Bart' s hydrops fetus was developed.

Bartter Syndrome ; diagnosis ; DNA ; blood ; Female ; Humans ; Hydrops Fetalis ; diagnosis ; Molecular Sequence Data ; Pregnancy ; Pregnancy Trimester, First ; Pregnancy Trimester, Second ; Pregnancy Trimester, Third ; Prenatal Diagnosis ; methods

OBJECTIVETo establish an in vitro model for the apatite crystal mineralization. To evaluate the influences of bovine serum albumin (BSA) and fluoride to the mineralization of apatite crystal.

METHODSThe model was constructed using cation selective membrane (CMV) and dialysis membrane. Double distilled water (DDW), BSA, 5, 20, 100 mg x L(-1) fluoride were added into the reaction space of the model. Reaction was carried out at 37 degrees C for 3 days under gentle stirring. The crystals were identified by scanning electron microscope (SEM) and X-ray diffraction (XRD).

RESULTSThe model was established successfully. When DDW and BSA were added respectively, the main component of the deposit was octacalcium phosphate (OCP), but the shape and size of the crystals differs from each other. When fluoride with different concentration were added, the main component of the crystal turned to rod-like and prism-like fluoroapatite (FAP) crystal. The size and crystallinity of the FAP increased with the increase of the fluoride concentration.

CONCLUSIONIt is an effective way to evaluate the influence factors of the apatite crystal mineralization by using the in vitro model.

Apatites ; Calcium Phosphates ; Crystallization ; Fluorides ; In Vitro Techniques ; Phosphates ; X-Ray Diffraction