Humans ; Salivary Glands

OBJECTIVE: To establish miniSTR fluorescent detection system with all detected fragments below 150 bp and to enhance the efficiency of detecting the degraded DNA samples. METHODS: All candidate primers were designed by Primer Premier 5 and screened by FastPCR 6.0. The miniSTR multiplex system was established by these selected loci labeling by four fluorescent dye. The parameters of PCR and primer concentrations were subsequently optimized. The electrophoresis was fulfilled under POP4 on 3100-Avant and the typing data was validated by standard DNA 9947A and 007. Fresh blood samples and difficult degraded DNA samples were tested to evaluate the usefulness of the system. RESULTS: All amplicons in the established miniSTR fluorescent detection system (D12ATA63, D2S1776, D1GATA113, D4S2408, D17S974, D20S482, D3S3053, Amelogenin, D6S474, D9S1122) were less than 150bp. The profile showed a balanced peak height without extra stutter by optimal protocol. Allele frequencies showed no deviations from Hardy-Weinberg equilibrium. The system showed accumulated probability of discrimination 0.999 999 983 and accumulated triplet excluding probability of paternity 0.996 8. It could detect corrupt muscle tissue, low copy number DNA samples and human tissues fixed by 40% formaldehyde solution for 12 days. CONCLUSION The miniSTR fluorescent detection system could be solely used for personal identification of degraded DNA samples or complementally used for paternity tests. And the system could enhance the ability of detecting the trace and degraded DNA.
DNA/chemistry* ; DNA Fingerprinting ; DNA Primers/genetics* ; Electrophoresis, Agar Gel ; Forensic Genetics ; Gene Frequency/genetics* ; Genetic Markers/genetics* ; Genetics, Population ; Humans ; Polymerase Chain Reaction/methods* ; Reference Standards ; Sequence Analysis, DNA/methods*

Objective To investigate the histomorphology,cellular biocompatibility in vitro and tensile force resistance property of fibrin-binding amniotic membrane(FBAM)and to explore its feasibility and superiority using as ocular surface graft.Methods After observing the morphology of FBAM under light microscope and scanning electron microscope,we did primary culture of corneal epithelial cells,taking monolayer fibrin-binding amniotic membrane as carrier,then observed cell growth overlay area.The elongation nature of FBAM was tested on electronic universal testing machine by calculating elastic modulus,using bilayer amniotic membrane(BAM)and monolayer amniotic membrane(MAM)as controls.Results The fresh FBAM was integrated as a whole,with structurally complete fibrin and amniotic membrane as well as tight binding of fibrin to amniotic membrane.The reserved FBAM was structurally complete,brittler than the fresh FBAM,while amniotic membrane still bound tightly to the fibrin,but under microscope the fiber reticular formation was fairly fuzzy.Single layer of epithelial cells was formed on FBAM after about seven-day culture in vitro.Disparity of cell growth overlay area was significant(P

Objective To investigate the surgical method of transnasal operations on the sphenoidal sinus and the middle fossa under nasal endoscope combined with microscope. Methods The operation was performed under nasal endoscope on the side of larger nasal cavity. The middle nasal turbinate was pushed outwards. Then the Handy's expander was inserted between the middle nasal turbinate and the nasal septum in order to get a wide visualization. The sphenoidal frontal wall was opened directly. The nasal endoscope and microscope were utilized in turn to complete the resection of lesions. Results The symptoms disappeared postoperatively in 10 cases of solitary sphenoidal sinusitis. The lesions of 6 cases of sphenoidal cyst and meningioma were all surgically removed on one session. Among 32 cases of pituitary adenoma, a total resection was carried out in 17 cases, a subtotal resection in 12 cases, and a partial resection in 3. After surgery, a supplemental X-knife radiosurgery was employed. Postoperative follow-up in the 48 cases for 0.5~3.5 years (mean, 2.5 years) showed no recurrence of sphenoidal cyst, sphenoidal sinusitis, or meningioma, and 3 cases of recurrence of pituitary adenoma. No intracranial infection, adhesion of nasal cavity, or nasal bleeding was noted. Conclusions Transnasal operations on the sphenoidal sinus and the middle fossa under nasal endoscope combined with microscope has advantages of mild invasion, little blood loss, short operation time, and good outcomes.

Adolescent ; Adult ; Diverticulum ; diagnostic imaging ; Female ; Humans ; Jugular Veins ; diagnostic imaging ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Young Adult

Objective To investigate the influence of thiamine deficiency(TD)at early pre- pathological lesion stage on cognitive function and the correlation between cognitive dysfunction and hippocampal neurogenesis.Methods TD mouse model was prepared by feeding a thiamine-depleted diet. Learning and memory functions of TD mice were tested with Y-maze.Hippoeampal neurogenesis was studied with bromodeoxyuridine(BrdU),proliferative cell nuclear antigen(PCNA),and Doublecortin(Dcx) immunohistochemical staining on the 7th(TD7),9th,14th and TD25th day.Results TD9 mice without pathological impairment and cholinergic nerve degeneration needed more times of training(22.3?2.2)in the learning test of Y maze compared with the controls(13.5?3.5).Correspondingly,the numbers of BrdU-positive ceils and the immunoreactivity of Dcx decreased significantly in the TD9 mice(19.8?0.4, 1537.2?50.2 vs 23.9?0.3,2688.9?127.9 pixels/mm~2).Thiamine re-administration reversed the declined hippocampal neurogenesis:the number of BrdU-positive cells was 23.6?1.9 and Dcx immunoreactivity was 2052.3?269.6 pixels/mm~2:the impaired learning ability was simultaneously restored,with the number of total training trial being 16.8?0.5.Conclusion The decreased hippocampal neurogenesis contributes to retarded learning ability at early pre-pathological lesion stage of TD.

Objective To analyze the clinical features of infantile cytomegalovirus(CMV) hepatitis with cholestasis and investigate intrahepatic cholestasis due to hepatocytic impairment caused by CMV infection.Methods Forty-seven children with CMV cholestatic he-patitis were divided into 2 groups according to the level of total bilirubin(TB):22 cases with serum TB lower than 136.8 ?mol/L(groupⅠ),and 25 cases with serum TB higher than 136.8 ?mol/L(groupⅡ).All children were treated with both gangciclovir and routine met-hods,and serum biochemistry were checked before and after treatment.SPSS 13.0 software was used to analyze the data.Results Forty-seven cases of infantile CMV cholestatic hepatitis had different degrees of jaundice,hepatosplenomegaly and abnormal liver functions.The differences of serum ALT and AST between the 2 groups had statistical significance,the levels of serum gamma glutamy transferase(GGT) and alkaline phosphatase(ALP) were lightly higher in groupⅡcompared with those in groupⅠ,but there were no statistical significance.TB,direct bilirubin(DB),ALT and AST were decreased in the 2 groups after treatment,GGT and ALP hadn′t decreased significantly after treatment.Conclusions CMV infection can injure hepatocytes and epithelials on each grade of bile duct,thus CMV hepatitis causes intrahepatic cholestasis.Cholestasis due to hepatocytic impairment deserves emphasis and intervention should be done as early as possible.Gangciclovir therapy for CMV infection manifest effective and safe in short-term.

Objective To investigate the role of directly constitutive activation of p38 mitogen-activated protein kinases(p38MAPKs)signaling in ?-globin gene expression and fetal hemoglobin(HbF)induction,and provide direct data for the relationship between phosphorylation of p38 and erythroid differentiation of human K562 erythroleukemia cells.Methods The human K562 erythroleukemia cells were transfected with pCDNA 3.1-MKK3(Glu)and pCDNA 3.1-MKK3(Ala)recombinant plasmids by lipofectamineTM 2000.Then,the stable cell lines overexpressing constitutively active p38 and constitutively inhibitive p38 activation were established by the addition of G418 to select single cell G418-resistant clones and identification with reverse transcriptase-polymerase chain reaction(RT-RCR)and Western blot assays,named K562-MKK3(Glu)and K562-MKK3(Ala)cells,respectively.Furthermore,the direct effects of constitutively active p38 on the ?-globin gene expression and HbF induction were analyzed by RT-PCR and benzidine staining,respectively.Results The results of RT-PCR and Western blot showed that there were no evident changes in the mRNA and protein levels of p38 for various cell models,but compared with K562,K562-vect,and K562-MKK3(Ala)cells,the phosphorylation of p38 and expression of ?-globin levels in K562-MKK3(Glu)cells were significantly up-regulated.The results of benzidine staining displayed that the mean percentages of positive cells stained by benzidine in K562,K562-vect,K562-MKK3(Ala),K562-MKK3(Glu)cells,and K562-MKK3(Glu)cells treated with SB203580 were(3.2?1.4)%,(3.7?1.2)%,(2.8?0.9)%,(32.6?5.3)%,and(7.8? 2.3)%(q = 7.56 P

1)which represented 340 genes and 171 down-regulated(SLR

OBJECTIVE: To perform the validation and analysis of forensic parameters of Goldeneye DNA ID 26Y system. METHODS: Based on the validation rules of Scientific Working Group on DNA Analysis Methods (SWGDAM), the kit was assessed from several parts, as test of PCR system, reproducibility, accuracy, and sensitivity, etc. And Y-STR loci of 517 unrelated healthy individuals from Eastern China were genotypes by this kit. The distribution and frequency of haplotype were calculated and forensic parameters of the kit were assessed. RESULTS: The complete profiles can be obtained even when the PCR reaction volume with 6.25 microL. And correct profile was obtained with DNA down to 125 pg. No reproducible peaks were detected with the DNA of common animals and microorganism with the kit. For the male-male mixture testing, average 70% of the minor alleles were obtained when the ratios of 1:19 and 19:1. For the male-female mixture testing, results showed that the sensitivity of the kit was no compromised with the addition of female samples. CONCLUSION The validation studies demonstrated that Goldeneye DNA ID 26Y system has good sensitivity and specificity, and suitable for mixture testing. The polymorphism of 26 Y-STR loci included in this kit are good for forensic application.
Alleles ; Animals ; Asian People/genetics* ; China ; Chromosomes, Human, Y ; DNA ; DNA Fingerprinting/standards* ; Female ; Forensic Genetics/methods* ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Reproducibility of Results ; Sensitivity and Specificity