A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.

ObjectiveTo evaluate the curative effect of reconstruction of the hand and foot defects with bones and tendons exposure using free medial sural artery perforator flap(MSAP). MethodsRadiographs of 2 cadavers injected with a modified lead oxide-gelatin mixture were digitally analyzed. Between April 2007 and December 2010, thirty-four patients with soft tissue defects in the distal limb were treated with the free MSAP flap transplantation. The sizes of the defect ranged 6 cm × 4 cm-13 cm × 8 cm, and the flaps ranged 7 cm× 5 cm-14 cm × 9 cm. These clinical cases included 25 hands and 9 feet, all of them with bones and tendons exposure.In these defects,twenty-two were clean,twelve got infections.In our cases, twenty-three flaps were nourished with single perforator vessel and else 11 with two;perforator vessel fifteen flaps were dissected one superficial vein to anastomose with that of the recipient sites in addition to accompanying vein anastomosis;The sensation of 9 flaps recovered the hands were reconstructed with cutaneous nerve anastomosis. ResultsA partition of the calf skin blood vessels,and three-dimensional reconstruction image of the sural artery were obtained.All flaps survived,five of them appeared partially violet and bubbles. Followed up 6-21 months, the cosmetic results were satisfactory and without apparent bulkiness.The flap colors were similar to recipient sites. The flap senses reconstructed with neural anastomosis recover to S2-S3. ConclusionThe new flap is very suitable to repair the soft tissue defect in the distal limbs,because the fairly constant perforator vessel,the reliable blood supply and the cosmetic shape of the MSAP flap are all advantages of it in addition to no damage to low leg chief artery and gastrocnemius.

Objective To provide anatomical landmarks with which to facilitate flap dissection,we studied the perforator artery of the dorsal forearm including its source,quantity,origination,caliber,variation and pedicle length. Methods Ten fresh cadavers were injected with a modified lead oxide-gelatin mixture,and three-dimensional graphics of the perforator vessels of the dorsal forearm were reconstructed with a computed tomography. In addition, twenty upper extremity specimens were injected with red latex via the axillary artery.The integument of the forearm was dissected,and perforators were identified,including type,course,size and location were documented.Surface areas were measured with Scion Image. Results The average number of the posterior interosseous artery cutaneous perforators in the dorsal forearm was (5±2),the average outer diameter of the perforator artories was (0.5 ± 0.1) mm,and the pedicle length was (2.5 ±0.2) cm.The average cutaneous vascular territory was (22.0 ± 15.0) cm2.The dorsal branch of the anterior interosseous artery dispersed on the wrist dorsum or the distal third of the dorsal forearm. It's average diameter was 0.8 mum. Conclusion The free transplantation of the posterior interosseous perforator artery flaps or rotary flap pedicled by the dorsal branch of the anterior interosseous artery for defect reconstruction are feasible.

Objective To determine the change of tumor bed volume during whole breast irradiation by repeated computed tomography scanning and to analyze the dosimetric impact of boost-planning on different CT images. Methods From July 2008 to Jan 2009, sixteen patients with early-stage breast cancer underwent breast conservative surgery (BCS) were enrolled in the study. All patients received whole breast irradiation and tumor bed boost, no adjuvant chemotherapy was given. Two additional CT scans were acquired in addition to the planning CT ( CT1 ), one in the course of radiotherapy ( CT2 ) and the other before the boost (CT3). Tumor beds were contoured in all CT images. Three-dimensional conformal radiotherapy planning for tumor bed boost was done on CT1 and CT3 respectively. Results The mean tumor bed volume on CT1, CT2 and CT3 were 49.5 cm3, 25.6 cm3 and 22. 2 cm3 ( F = 5. 63, P = 0. 007 ),respectively. Further analysis found statistically significant difference between CT1 and CT2 ( q = 0. 03, P =0. 010), CT1 and CT3 ( q = 0. 01, P = 0. 004), but not between CT2 and CT3 ( q = 1.00, P = 0. 333 ). The average reduction of tumor bed volume from CT1 to CT3 was 43.4%. A reduction of 20% or above was found in 88% of the patients ( n = 14), 50% or above in 38% of the patients (n = 6). In the boost-planning, the volume of the ipsilateral breast receiving 100% prescribed dose (V100%) on CT1 and CT3 was 183.5 cm3 and 144. 5 cm3, respectively ( t = 3.06, P = 0. 008 ). Conclusions Volume of tumor bed is dynamically reduced in the course of whole breast irradiation after BCS, with more important reduction in the early weeks after the beginning of irradiation. A second CT scan before tumor bed boost is warranted.

The metabonomics method was used to study the intervention effect of Psoraleae Fructus and Myristicae Semen in "Ershen pill" on the changes in serum endogenous metabolites in spleen-kidney Yang deficiency diarrhea rats before and after processing, screen out differentiated metabolites related to spleen-kidney Yang deficiency diarrhea and explore the metabolic patterns related to spleen-kidney Yang deficiency diarrhea and the processing synergy mechanism of Psoraleae Fructus and Myristicae Semen in "Ershen pill". Efforts were made to detect SOD and MDA of each group, test rat serum metabolic fingerprints in different stages by using GC-MS, analyze by PCA and PLS-DA methods and screen out potential biomarks through VIP and t test. The results revealed that "Ershen pill" could enhance the level of SOD and decrease the level of MDA and identified 10 differentiated metabolites related to spleen-kidney Yang deficiency diarrhea. Compared with the model group, all of metabolites recovered to varying levels after being intervened with "Ershen pill", with the best effect shown in the "Ershen pill" IV group (salt-processed Psoraleae Fructus + bran-roasted Myristicae Semen). It is speculated that that Psoraleae Fructus and Semen Myristicae in "Ershen pill" show a synergistic effect by inhibiting peroxide, improving aglucolipid, amino acids and energy metabolism, with multiple target sites.
Animals ; Chemistry, Pharmaceutical ; Diarrhea ; drug therapy ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Energy Metabolism ; drug effects ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Metabolomics ; Myristicaceae ; chemistry ; Psoralea ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spleen ; drug effects ; metabolism ; Yang Deficiency ; drug therapy ; metabolism

To explore the growth and development and analyze the quality of the parthenocarpy fruit induced by exogenous hormones of Siraitia grosvenorii. the horizontal and vertical diameter, volume of the fruit were respectively measured by morphological and the content of endogenous hormones were determined by ELISA. The size and seed and content of mogrosides of mature fruit were determined. The results showed that the fruit of parthenocarpy was seedless and its growth and development is similar to the diploid fruit by hand pollination and triploid fruit by hand pollination or hormones. But the absolute value of horizontal and vertical diameter, volume of parthenocarpy fruit was less than those of fruit by hand pollination, while triploid was opposite. The content of IAA, ABA and ratio of ABA/GA was obviously wavy. At 0-30 d the content of IAA and ABA of parthenocarpy fruit first reduced then increased, content of IAA and GA parthenocarpy fruit was higher than that of fruit by hand pollination. Mogrosides of parthenocarpy fruit was close to pollination fruit. Hormones can induce S. grosvenorii parthenocarpy to get seedless fruit and the fruit shape and size and quality is close to normal diploid fruit by hand pollination and better than triploid fruit by hormone or hand pollination.
Cucurbitaceae ; chemistry ; drug effects ; genetics ; growth & development ; Diploidy ; Fruit ; chemistry ; genetics ; growth & development ; Plant Growth Regulators ; pharmacology

The relationship between maternal nutrient intake and fetal size or infant growth was studied in 1956 pregnant women, 599 parturients and 1043 lactating women, 318 non-pregnant women included as controls. The study was conducted in eight regions that were representative of all geographical areas of China. The diet was comprised primarily of cereal products with 70% to 85% of the zinc intake derived from plant sources. Women in the third trimester of pregnancy, parturients and lactating women consumed more food than non-pregnant women or women in the first two trimesters of pregnancy. Total energy, protein and iron intakes met the recommended allowances for each stage of reproduction. Calcium and zinc intakes, however, were 50% and 47% of the amount recommended, respectively. Only 7.2% of the women exceeded two-thirds of the recommended zinc intake. The mean intake of zinc was 6.5mg to 9.0 mg each day among all the subjects. Correlation and stepwise regression analysis showed that maternal zinc intake was a predictor factor for fetal dimensions and birthweight. The results of this study show that fetal growth and birthweight are directly related to maternal zinc intake among Chinese women, and that there is no relationship between maternal zinc intake during lactation and infant height, weight, or weight gain from birth.

Objective To explore the relationship between size,shape and function of the left atrium appendage (LAA) and its thrombosis in patients with atrial fibrillation (AF) by transesophageal echocardiography (TEE) to provide evidence for clinical risk assessment,prognosis evaluation and treatment guidance.Method Length,diameter,end-diastolic volume (EDV) and ejection velocity (PEV) of LAA were measured in 41 patients with AF,and thrombus in LAA was detected by TEE.Results Thrombus in LAA was detected in seven of 41 patients of AF (17%).No significant difference in size and EDV was found between the patients with and without thrombus,but there was significant difference in PEV between them (P

OBJECTIVETo develop a multiplex real-time PCR for the detection of Salmonella invasion protein A gene (invA), enterotoxigenic Escherichia coli (ETEC) heat-labile I enterotoxin gene (elt), and Shigella or enteroinvasive E. coli (EIEC) invasive plasmid antigen H gene (ipaH).

METHODSUnder the optimized reaction conditions of the multiplex real-time PCR, invA, elt, and ipaH were determined in 10-fold series of dilution of DNA extracted from Salmonella enterica serovar Typhimurium, ETEC 44815 strain and Shigella F301 strain. The three genes were examined in 90 fecal samples from diarrhea patients using the multiplex real-time PCR. When PCR-positive samples were found, the target strains were isolated and identified.

RESULTSThe detectable concentration for this multiplex real-time PCR was 10 CFU/microl for Shigella F301 strain, 10(2) CFU/microl for S. enterica serovar Typhimurium and ETEC 44815 strain, respectively. Out of 90 fecal samples from diarrhea patients, thirteen were found positive for elt gene (14.4%), and five were found positive for ipaH gene (5.6%). Three E. coli strains positive for elt gene and four E. coli strains positive for ipaH gene were isolated successfully from the PCR-positive samples mentioned above. The detection of invA, elt and ipaH genes was completed in 10 h, which included an enrichment period of 6 h.

CONCLUSIONThe multiplex real-time PCR assay can detect invA, elt, ipaH simultaneously in a single reaction, moreover, it can detect for virulence genes in strains of Salmonella, ETEC, and Shigella or EIEC and screen these pathogens in fecal specimens from patients with diarrhea with a high specificity.

DNA, Bacterial ; analysis ; Diarrhea ; microbiology ; Escherichia coli ; genetics ; Feces ; microbiology ; Humans ; Polymerase Chain Reaction ; methods ; Salmonella ; genetics ; Shigella ; genetics

OBJECTIVERett syndrome (RTT) is a severe childhood neurodevelopmental disorder mainly affecting females. The pathogenic gene is located at Xq28, which codes for the methyl-CpG-binding protein 2. MECP2 gene is affected by X chromosome inactivation (XCI). The different XCI patterns of females could affect the expression ratios of pathogenic gene, causing changes in clinical symptoms. In order to understand the XCI patterns in RTT patients and the relationship between XCI pattern, genotype and phenotype, the XCI patterns in patients with RTT and their mothers, the parental origin of the priority inactive X chromosome in RTT, and the relations of XCI patterns with genotype and phenotype in RTT cases were analyzed.

METHODSGenomic DNA was extracted from peripheral blood of 55 cases with RTT (52 with MECP2 mutations, 3 without mutations), 53 mothers of RTT cases and 48 normal female controls. DNA was digested with methylation sensitive restriction endonuclease Hpa II. Then the undigested and digested DNAs were amplified via PCR for the first exon of human androgen receptor (AR) gene. PCR products were analyzed by Genescan.

RESULTSThe heterozygotic rates of AR gene were 82%, 77% and 83% in RTT patients, mothers and controls, respectively. XCI distribution pattern of RTT was different from that of the mothers and control, P < 0.05. More mothers and controls than RTT patients were in the area of XCI 50:50 - 59:41. The differences between them were statistically significant (P < 0.05). No significant difference in XCI distribution patterns between mothers and the control groups was found (P > 0.05). Non-random XCI rates in the areas of XCI >or= 65:35 and >or= 80:20 were 53.35% and 17.8%, respectively, in RTT patients, compared with the mothers group (36.6%, 7.3%) and control group (35%, 10%), it was higher in RTT patients, but the difference was not statistically significant (P > 0.05). In 18 of 21 cases with XCI >or= 65:35, the priority inactive X chromosome was of paternal origin (85.7%). Variable XCI patterns were observed in the same gene mutation patients. The highly skewed XCI as well as the random XCI were found in patients with mild, severe and typical phenotype. The rate of highly skewed XCI in atypical patients was higher than that in typical RTT patients. The rate of highly skewed XCI in T158M was higher than the other type mutations. No highly skewed XCI was observed in cases with R133C mutation.

CONCLUSIONThe XCI distribution pattern of RTT patients was different from that of RTT mother and control groups. There was no significant difference in XCI distribution patterns between mothers and the control groups. It was not a main genetic pattern in RTT that mothers as the carriers to transmit the pathogenic gene to the patients. Non-random XCI was not the main XCI pattern in RTT patients. The priority inactive X chromosome was mainly of paternal origin. XCI could modify the clinical phenotype of RTT, but had limitations in explaining all the phenotypes manifested in RTT cases.

Adolescent ; Adult ; Child ; Child, Preschool ; Chromosomes, Human, X ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Heterozygote ; Humans ; Mothers ; Mutation ; Phenotype ; Receptors, Androgen ; genetics ; Rett Syndrome ; diagnosis ; genetics ; X Chromosome Inactivation ; genetics