Objective To develop a sensitive and reliable RP-HPLC method for the simultaneous determination of nine nucleosides including uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine from Fritillaria taipaiensis P. Y. Li that had been cultivated in different producing areas; To compare the contents of these nucleosides from different producing areas. Methods The analysis was performed on a Venusil MP C18 (2) column (4.6 mm×250 mm, 5 μm) with a gradient of methanol-water at a flow rate of 1.0 mL/min;the detective wavelength was set at 260 nm; the column temperature was set at 35 ℃. Results Uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine were obtained in the good linear range of 0.269 5–16.17 μg/mL, 0.132 1–7.927 5 μg/mL, 0.095 5–5.73 μg/mL, 1.16–69.6 μg/mL, 0.48–28.8 μg/mL, 0.571 5–57.15 μg/mL, 0.526–52.6 μg/mL, 3.307 5–198.45 μg/mL, 0.530 5–31.83 μg/mL, respectively (r≥0.999 5);the recovery was in the range of 96.49%–101.65%(RSD≤2.92%). Conclusion The contents of the nine nucleosides from different producing areas have differences. Fritillaria taipaiensis P. Y. Li from Xianyi Village, Chengkou County, Chongqing City, whether the cultivated ones or wild ones, contain the highest level of nucleosides. The established method can provide references for the perfection of quality standard for Fritillaria taipaiensis P. Y. Li.

OBJECTIVETo establish a method based on molecular beacon real-time PCR for detecting single nucleotide polymorphisms (SNP) in codon 72 of scar-related p53 gene.

METHODSTwo fluorescence-labeled molecular beacon probes were synthesized targeting CCC/CGC SNP of p53 codon 72. The genomic DNA was extracted from the peripheral blood of 28 patients with keloid, and the CCC/CGC SNP of P53 gene codon 72 were assayed with molecular beacon real-time PCR. The results of SNP typing were compared with the results of reverse dot hybridization and confirmed by direct DNA sequencing.

RESULTSThe goodness of fit of this method was 100% in comparison with direct DNA sequencing, higher than that of reverse dot hybridization.

CONCLUSIONMolecular beacon real-time PCR is suitable for rapid clinical detection of SNPs in p53 gene.

Base Sequence ; Codon ; genetics ; Humans ; Keloid ; genetics ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Real-Time Polymerase Chain Reaction ; methods ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo study the clinical efficacy of three kinds of hybrid bioartificial liver support systems (HBLSS) in treating chronic severe hepatitis.

METHODSA bioartificial liver support system (BAL), comprising porcine hepatocytes and fiber tube style bioreactor, was constructed. Then three kinds of HBLSS were constructed: Molecular absorbent recirculating system (MARS) plus BAL; slow plasma exchange (SPE) plus continuous hemodiafiltration (CHDF) and BAL; and SPE plus hemoperfusion (HP) and BAL. One hundred-twenty patients in middle or late stages of chronic severe hepatitis were enrolled in this study. They were randomly divided into 6 groups: H1 group was treated with BAL+MARS, H2 with BAL+SPE+CHDF and H3 with BAL+SPE+HP (as treatment groups); C1 group was treated with MARS, C2 with SPE+CHDF and C3 with SPE+HP (as control groups). The changes in the clinical symptoms, in the hepatic encephalopathy stages, and in the serum total bilirubin (TBIL), the serum albumin (ALB), the prothrombin activities (PTA), endotoxin, ammonia, creatinine and a-fetal protein (AFP) were all observed before the treatment, right after it and 72 hours later. The improving and curing rates and the rates of side effect occurrences in each group were observed.

RESULTSIn all 6 groups, the patients' clinical symptoms ameliorated; their TBIL, endotoxin and ammonia levels decreased (P<0.05), and their PTA and AFP levels lowered significantly (P<0.05). But in the H1, H2 and H3 groups they were more distinctive than in the control groups. In H1 and H2 groups creatinine and ammonia levels were decreased more significantly than in the H3 group (P<0.05). The improving and curing rates of each group were 65 % (13/20), 60% (12/20), 45% (9/20), 45% (9/20), 40% (8/20) and 20% (4/20) respectively. No serious side effects were observed during the treatment.

CONCLUSIONIn treating middle and late stage chronic severe hepatitis, the measures used in H1, H2 and H3 are better than those in C1, C2 and C3. Furthermore, H1 and H2 treatments can ameliorate hepatic and renal functions, prevent the development of multiple organ dysfunction syndrome, and are better than those used in H3.

Adult ; Aged ; Animals ; Bioreactors ; Critical Illness ; Female ; Hemodiafiltration ; Hepatic Encephalopathy ; blood ; therapy ; Hepatitis, Viral, Human ; therapy ; Humans ; Liver ; cytology ; Liver Failure, Acute ; therapy ; Liver, Artificial ; Male ; Middle Aged ; Plasma Exchange ; Swine

Objective To investigate the carrier ratio and the genotype of thalassemia among students of secondary school in Chongzuo, Guangxi. Methods From June 10-20,2008 among 7 regions of Chongzuo, 1 secondary school was randomly chosen from each region, and the number of student volunteers was determined by 0.5‰ proportion of the local population size. 1097 students were screened, including 515 boys and 582 girls of 12-16 year olds. Among them, 968 cases were Zhuang (438 boys and 530 girls) 128 cases were Han (76 boys and 52 girls) and one case was Yao nationalities (boy). Analysis of blood cells was detected by Cell Dyn 1700 automatic hemocyte analysator while hemoglobin F (HbF) and hemoglobin A2 (HbA2) were detected by hemoglobin autoanalyse variant. Among those with HbA2≥ 4% that belonged to β-thalassemia before α and β-thalassemia gene were analyzed to identify the genotypes. IfHbA2 was <4% but MCV≤80 fl, α-thalassemia gene was analyzed. Results Among 1097 cases,218 wereα-thalassemia (19.87%), 50 were β-thalassemia (4.56%) and 13 were combination of α β-thalassemia (1.19%). The overall detected ratio was 25.62%. 133 cases with thalassemia were boys (25.83%) and 148 were girls (25.43%) with no significant difference(P>0.05). 255 cases of thalassemia were Zhuang (26.34%), and 25 were Han nationality (19.52%). The detected ratio among Zhuang nationality was higher than in Hun nationality and with significant difference statistically (P<0.01). 3 kinds of deletion (-α3.7/,-α<4.2>/, --SEA/) and another 3 kinds of non-deletion (αα CS/, αα WS/, αα QS/) α-thalassemia genotype were identified, with a higher rate of ααWS/. Among the β-thalassemia genotype, CD41-42 appeared the most common genotype. MCV of thalassemia was lower than in the controls, with significant difference (P<0.01). 78-90 fl of α-thalassemia was detected from the MCV specimen. If taken MCV<79 fl as the positive phenotype of thalassemia, 32 cases were misdiagnosed. The rate of missed diagnosed cases was 2.97%. Conclusion Rate of thalassemia carder among students of secondary school in Chongzuo, Guangxi was considered to be high, especially those belonged to Zhuang nationality were higher than the Hans. The carrier rate of ααWS/was higher, with CD41-42 the most common genotype.

Background/Aims: Diagnosis of isolated laryngopharyngeal reflux symptoms (ILPRS), ie, without concomitant typical reflux symptoms (CTRS), remains difficult. Mean nocturnal baseline impedance (MNBI) reflects impaired mucosal integrity. We determined whether esophageal MNBI could predict pathological esophagopharyngeal reflux (pH+) in patients with ILPRS. Methods: In this cross-sectional study conducted in Taiwan, non-erosive or low-grade esophagitis patients with predominant laryngopharyngeal reflux symptoms underwent combined hypopharyngeal multichannel intraluminal impedance-pH monitoring when off acid suppressants. Participants were divided into the ILPRS (n = 94) and CTRS (n = 63) groups. Asymptomatic subjects without esophagitis (n = 25) served as healthy controls. The MNBI values at 3 cm and 5 cm above the lower esophageal sphincter (LES) and the proximal esophagus were measured. Results: Distal but not proximal esophageal median MNBI values were significantly lower in patients with pH+ than in those with pH– (ILPRS in pH+ vs pH–: 1607 Ω vs 2709 Ω and 1885 Ω vs 2563 Ω at 3 cm and 5 cm above LES, respectively; CTRS in pH+ vs pH–: 1476 vs 2307 Ω and 1500 vs 2301 Ω at 3 cm and 5 cm above LES, respectively, P < 0.05 for all). No significant differences of any MNBI exist between any pH– subgroups and healthy controls. The areas under the receiver operating characteristic curve in the ILPRS group were 0.75 and 0.80, compared to the pH– subgroup and healthy controls (P < 0.001 for both), respectively. Interobserver reproducibility was good (Spearman correlation 0.93, P < 0.0001). Conclusion Distal esophageal MNBI predicts pathological reflux in patients with ILPRS.

The full-length P32 gene and the truncated P32 gene (MP-32) were amplified from the recombinant plasmid pMD-P32 by polymerase chain reaction (PCR) and cloned into pcDNA3. 1(+) and pcDNA3.1-CpG respectively. The recombinant plasmids (pcDNA3.1-P32, pcDNA3.1-CpG-P32 and pcDNA3. 1-CpG-MP32) were transfected into BHK-21 cells by using lipofectin. The expressed P32 protein was confirmed by indirect immunofluorescence assay (IFA). The BALB/c mice were immunized with these recombinant plasmids by intramuscular injection. The specific antibodies aginst CPV were detected by ELISA kit weekly. The murine splenic T lymphocyte subgroups CD4+ and CD8+ were detected by flow cytometry. Results showed that the P32 protein was expressed successfully in vitro. After 2 weeks post im munization, the specific IgG antibodies against CPV were detected in the vaccinated mice. The percentage of CD4+ /CD8+ T cells was significantly higher than that of the control. In conclusion, these constructed eukaryotic vectors could induce humoral and celluar immune responses in mice.
Animals ; Antibodies, Viral ; blood ; Capripoxvirus ; genetics ; immunology ; Cell Line ; CpG Islands ; Cricetinae ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; immunology ; T-Lymphocyte Subsets ; immunology ; Vaccines, Synthetic ; immunology ; Viral Envelope Proteins ; immunology ; Viral Vaccines ; immunology

OBJECTIVETo assess the accuracy of detection by automated breast volume scanner (ABVS) in diagnosis of high-risk and small breast lesions.

METHODSOne hundred and twelve patients with solid high-risk and small breast lesions were identified by ABVS. The patients were divided into benign lesion group and cancer group after pathological examination. The clinicopathological findings and ultrasonographic features of the lesions were compared.

RESULTSAmong the 112 lesions there were 49 benign and 63 malignant lesions. The mean size on ABVS and pathology were (1.59 ± 0.52) cm and (1.52 ± 0.58) cm. There was no significant difference in tumor sizes determined by ABVS and pathology (P = 0.194). The mean age of patients with benign lesions was (38.5 ± 7.4) years and that of malignant lesions was (52.4 ± 13.6) years, showing a significant difference between the two groups (P < 0.001) . The mass shape, orientation, margin, lesion boundary, echo pattern, calcification, BI-RADS category and retraction phenomenon were significantly different of the malignant and benign masses (P < 0.05). But there was no significant difference in the location of lesions and posterior acoustic features (P > 0.05) . Retraction phenomenon was significantly associated with pathological type and histologic grade of the breast cancer (P < 0.01). The specificity, sensitivity and accuracy of retraction phenomenon were 100% (46/46), 73.0% (46/63), and 84.8% (95/112), respectively.

CONCLUSIONSABVS provides advantages of better size prediction of high-risk and small breast lesions. Furthermore, the retraction phenomenon in coronal plane shows high specificity and sensitivity in detecting breast cancer.

Adult ; Age Factors ; Aged ; Aged, 80 and over ; Breast Neoplasms ; diagnostic imaging ; pathology ; Carcinoma, Ductal, Breast ; diagnostic imaging ; pathology ; Female ; Fibroadenoma ; diagnostic imaging ; pathology ; Humans ; Image Enhancement ; methods ; Image Interpretation, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Middle Aged ; Retrospective Studies ; Sensitivity and Specificity ; Tumor Burden ; Ultrasonography, Mammary ; methods ; Young Adult

DNA, Mitochondrial ; genetics ; Genetic Testing ; Humans ; Mitochondria ; Mutation ; RNA, Transfer, Leu