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Objective: To observe the effects of taurine on hemorheology and oxidative stress of diabetic rats., Methods: 40 rats were divided into control group, diabetes group and treatment group at random. Diabetic model was reproduced by intraperitoneal injection of streptozotocin. After having been treated with taurine for 8 weeks, glycosylated hemoglobin (HbA1c) and the serum contents of glucose, superoxide dismutase(SOD), malondialdehyde (MDA) were measured. The changes of hemorheology in different groups were detected respectively., Results: Compared with control group, the content of glucose, MDA and HbA1c in diabetic rats was increased, the activity of SOD was decreased, the levels of whole blood viscosity and the aggregation index of red blood cells and hematocrit were increased and RBC deformability index was decreased in diabetic rats. Moreover, taurine was able to apparently reduce high blood glucose and HbA1c (P < 0.05), markedly elevated the activity of SOD, lowered the content of MDA (P < 0.01); and taurine also could significantly reduce the levels of whole blood viscosity and the aggregation index of red blood cells and hematocrit in the meanwhile, and increase RBC deformability index (P < 0.05 or P < 0.01)., Conclusion: Taurine could enhance the ability of oxidation resistance, improve blood rheology property in diabetic rats, at the same time it could be beneficial to prevent and cure the development of diabetic blood vessel complication.

Allelochemicals released from root exudates or decaying residues of plants play diversified roles in ecological interactions of plant-pathogen. The objective of this work was to evaluate the allelopathic effect of an externally supplied tannic acid on soil-borne in vitro Fusarium oxysporum f. sp. niveum. Results showed that the tannic acid decreased the growth of the fungus up to 9.5% at 800 mg l(-1). Conidial germination was reduced by 52.3% in comparison with the control. However, sporulation and mycotoxin production by the fungus were stimulated. The activity of pectinase and proteinase were initially increased and finally decreased with increase in concentrations of tannic acid. Tannic acid served as an ecological allelochemical, repressing the growth of the pathogen.

Aim: To study the potential value and specificity of plasma miR-216a as a marker for pancreatic injury., Methods: Two rat models were applied in this article: L-arginine-induced acute pancreatitis was used as one model to explore the potential value of plasma miR-216a for detection of pancreatic injury; nonlethal sepsis induced in rats by single puncture cecal ligation and puncture (CLP) was used as the other model to evaluate the specificity of plasma miR-216a compared with two commonly used markers (amylase and lipase) for acute pancreatitis. Plasmas were sampled from rats at indicated time points and total RNA was isolated. Real-Time Quantitative reverse transcriptase-polymerase chain reaction was used to quantify miR-216a in plasmas., Results: In the acute pancreatitis model, among five time points at which plasmas were sampled, miR-216a concentrations were significantly elevated 24 h after arginine administration and remained significantly increased until 48 h after operation (compared with 0 h time point, P < 0.01, Kruskal-Wallis Test). In the CLP model, plasma amylase and lipase, two commonly used biomarkers for acute pancreatitis, were significantly elevated 24 h after operation (compared with 0 h time point, P < 0.01 and 0.05 respectively, Pairwise Bonferroni corrected t-tests), while miR-216a remained undetectable among four tested time points., Conclusion: Our article showed for the first time that plasma miR-216a might serve as a candidate marker of pancreatic injury with novel specificity.

Aims and Background: Transcriptional silencing induced by hypermethylation of CpG islands in the promoter regions of genes is believed to be an important mechanism of carcinogenesis in human cancers including gastric cancer. A number of reports on methylation of various genes in gastric cancer have been published, but most of these studies focused on cancer tissues or only a single gene. In this study, we determined the promoter hypermethylation status and mRNA expression of 4 genes: p16, Runx3, DAPK and CHFR., Methods: Methylation-specific polymerase chain reaction (MSP) was used to determine the methylation status of p16, Runx3, DAPK and CHFR gene promoters in cancer and adjacent normal gastric mucosa specimens from 70 patients with gastric cancer, as well as normal gastric biopsy samples from 30 people without cancer serving as controls. In addition, the mRNA expression of p16, Runx3, DAPK and CHFR was investigated in 34 gastric cancer patients by RT-PCR. Bisulfite DNA sequence analysis was applied to check the positive samples detected by MSP., Results: When carcinoma specimens were compared with adjacent normal gastric mucosa samples, a significant increase in promoter methylation of p16, Runx3, DAPK and CHFR was observed, while all 30 histologically normal gastric specimens were methylation free for all 4 genes. The methylation rate of the 4 genes increased from normal stomach tissue to tumor-adjacent gastric mucosa to gastric cancer tissue. Concurrent methylation in 2 or more genes was found in 22.9% of tumor-adjacent normal gastric mucosa and 75.7% of cancer tissues. No correlation was found between hypermethylation and other clinicopathological parameters such as sex, age, and tumor location. However, the frequency of DAPK and CHFR methylation in cancer tissues was significantly associated with the extent of differentiation and lymph node metastasis (P < 0.05) and the frequency of Runx3 methylation was significantly associated with tumor size (P < 0.05). Weak expression and loss of expression of the 4 genes was observed in cancer tissues and was significantly associated with promoter hypermethylation (P < 0.05)., Conclusions: Promoter hypermethylation of p16, Runx3, DAPK and CHFR is frequent in gastric cancer. DAPK and CHFR promoter hypermethylation may be an important help in evaluating the differentiation grade and lymph node status of gastric cancer. Weak gene expression and loss of gene expression due to promoter hypermethylation may be a cancer-specific event.

Group A rotavirus are the most frequently detected viral agent associated with the acute diarrhea in calves. In order to investigate the situation of rotavirus strains circulating in diary farms, a total of 117 fecal specimens were collected from diarrhea calves under 4 weeks-age on Yinluo diary farm in Daqing region in China from 2008 to 2009. Ten specimens were detected to be positive by a Rotavirus Group A Diagnostic Kit, which confirmed that the rotavirus was important viral agent associated with diarrhea in this diary farm. Based on the new classification system, G10P[11] genotype was determined in rotavirus positive samples. Sequence and Phylogenetic analysis indicated DQ-75 strain was introduced into our country with imported bovine.

In this paper, various N-trimethyl chitosan (TMC) of different molecular-weights (approximately 100 KD, approximately 200 KD, and approximately 400 KD, respectively) with the approximately degree of quartenization (DQ) of 40% were successfully synthesized. In vitro cytotoxicity of TMC solution showed the dependence of TMC concentration from 20 microg/ml to 500 microg/ml on the relative cell activity. Molecular weight of TMC did not greatly affect the cytotoxicity of TMC against HEK293 and L929 cells. TMC nanoparticles and alginate modified TMC nanoparticles were prepared by the ionic gelation method. Subsequently, we investigated the properties of TMC nanoparticles and alginate modified TMC nanoparticles intending for oral delivery of antigens. Molecular weight of TMC did not affect the loading capacity (LC) and in vitro release behavior of TMC nanoparticles. However, BSA concentration and alginate modification have strongly effect on properties of TMC nanoparticles (particle size; surface charge; loading efficiency and loading capacity). In vitro release behavior indicated that alginate modification could efficiently decrease initial burst release and extend release time in phosphate buffer (PBS, pH 7.4) and acidic solution (0.1 M HCl, pH = 1) at 37 degrees C. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) assay showed that alginate modification could effectively improve the stability of TMC nanoparticles and protect BSA from degradation or hydrolysis in acidic condition for at least 2 h.

The transcription factor (TF) is a protein that binds DNA at specific site to help regulate the transcription from DNA to RNA. The mechanism of transcriptional regulatory can be much better understood if the category of transcription factors is known. We introduce a system which can automatically categorize transcription factors using their primary structures. A feature analysis strategy called "mRMR" (Minimum Redundancy, Maximum Relevance) is used to analyze the contribution of the TF properties towards the TF classification. mRMR is coupled with forward feature selection to choose an optimized feature subset for the classification. TF properties are composed of the amino acid composition and the physiochemical characters of the proteins. These properties will generate over a hundred features/parameters. We put all the features/parameters into a classifier, called NNA (nearest neighbor algorithm), for the classification. The classification accuracy is 93.81%, evaluated by a Jackknife test. Feature analysis using mRMR algorithm shows that secondary structure, amino acid composition and hydrophobicity are the most relevant features for classification. A free online classifier is available at http://app3.biosino.org/132dvc/tf/.

Genetic variation and evolutionary demography of the shrimp Fenneropenaeus chinensis were investigated using sequence data of the complete mitochondrial control region (CR). Fragments of 993 bp of the CR were sequenced for 93 individuals from five localities over most of the species' range in the Yellow Sea and the Bohai Sea. There were 84 variable sites defining 68 haplotypes. Haplotype diversity levels were very high (0.95 ± 0.03-0.99 ± 0.02) in F. chinensis populations, whereas those of nucleotide diversity were moderate to low (0.66 ± 0.36%-0.84 ± 0.46%). Analysis of molecular variance and conventional population statistics (F(ST) ) revealed no significant genetic structure throughout the range of F. chinensis. Mismatch distribution, estimates of population parameters and neutrality tests revealed that the significant fluctuations and shallow coalescence of mtDNA genealogies observed were coincident with estimated demographic parameters and neutrality tests, in implying important past-population size fluctuations or range expansion. Isolation with Migration (IM) coalescence results suggest that F. chinensis, distributed along the coasts of northern China and the Korean Peninsula (about 1000 km apart), diverged recently, the estimated time-split being 12,800 (7,400-18,600) years ago.

It is well known that spinal motoneurons below a spinal transection become supersensitive to a systemic administration of serotonin (5-HT) precursors, such as 5-hydroxytryptophan. This supersensitivity has been implicated in both the process of functional recovery following chronic lesions, and also in the development of symptoms such as hyperreflexia and spasticity. However, the mechanisms of this denervation supersensitivity are still largely unknown. In this study we have investigated the changes in 5-HT2A receptor immunoreactivity following chronic spinal transections at the level of the sacrocaudal spinal cord. The results show that in the spinalized rats the immunoreactivity of 5-HT2A receptors below the lesion is dramatically increased in the motoneuron soma and its proximal dendritic territory, most likely also in their distal dendritic territory, to a level 3-5-fold higher than that of sham-operated rats. We also found a small number of intraspinal 5-HT neurons and clusters of 5-HT fibers and their varicosities in the spinal cord caudal to the lesion, which may provide an intrinsic source of 5-HT to act upon the upregulated 5-HT2A receptors. These results indicate that the upregulation of 5-HT2A receptors at least partly underlies the 5-HT denervation supersensitivity of spinal motoneurons after a complete spinal transection., (Copyright 2010 Elsevier B.V. All rights reserved.)

Background and Objective: Transcriptional silencing induced by CpG island methylation is believed to be one of the important mechanisms of carcinogenesis. Checkpoint with fork head-associated and ring finger (CHFR) governs the transition from prophase to prometaphase in response to mitotic stress. This study was to analyze the relationship between the methylation of CHFR gene and the clinicopathologic features of gastric cancer, and the difference of results between methylation-specific polymerase chain reaction (MSP) and combined bisulfite restriction analysis (COBRA) in detecting aberrant methylation of CHFR gene in gastric cancer., Methods: Both MSP and COBRA methods were used to detect the promoter methylation of CHFR gene in gastric cancer specimens from 64 patients. The relationship between methylation status of CHFR gene and the clinicopathologic features of gastric cancer were analyzed using SPSS16.0., Results: The methylation rates of CHFR gene promoter were significantly higher in gastric cancer samples than in the corresponding paracancer normal gastric mucosa by MSP (51.6% vs. 18.8%, P < 0.001). However, there was no significant correlation between methylation status of CHFR gene and the clinicopathologic parameters of gastric cancer, including age, gender, tumor size, clinical stage, Borrman type, tumor invasion depth, differentiation, and lymph node metastasis (P > 0.05). Aberrant methylation of the CHFR gene was detected in 27 (42.2%) of the 64 specimens of gastric cancer using COBRA, which did not significantly differ from that using MSP (P > 0.05)., Conclusions: Aberrant methylation of the CHFR gene is a frequent event in the carcinogenesis of gastric cancer. Detecting the methylation of CHFR gene in gastric mucosa may conduce to the diagnosis of gastric cancer. No difference was found between MSP and COBRA in detecting promoter methylation of CHFR gene in gastric cancer.

Aim: To observe the differences of damage patterns of retinal nerve fiber layer (RNFL) between acute and chronic intraocular pressure (IOP) elevation in primary angle closure glaucoma (PACG) using optical coherence tomography (OCT)., Methods: Twenty-four patients (48 eyes) with unilateral acute PACG (APACG) attack in the 6 months after admission and 36 patients (64 eyes) with chronic PACG (CPACG) were included in this prospective study. For all cases, IOP has been controlled under 21mmHg after treatment. Using stratus OCT, the RNFL thickness was assessed in eyes with PACG within 3 days, 2 weeks, 1, 3 and 6 months after IOP was controlled. Repeated measures ANOVA was used to examine the changes of RNFL thickness at different time after IOP being controlled in both acute attack eyes and unaffected fellow eyes of APACG and eyes with CPACG., Results: The mean RNFL thickness for the APACG-attacked eyes increased significantly within 3 days (121.49±23.84)µm after acute onset and then became thinner along with time [(107.22±24.72)µm at 2 weeks,(93.58±18.37)µm at 1 month, (84.10±19.89)µm at 3 months and (78.98±19.17)µm at 6 months]. In APACG-attacked eyes, there were significant differences of average RNFL thickness at 5 different times after IOP was controlled (P<0.001). In the APACG unaffected fellow eyes and CPACG eyes, there were no significant differences in mean RNFL thickness at 5 different times(F=0.450, P=0.104 in APACG unaffected fellow eyes and F=1.558, P=0.200 in CPACG eyes). There was significant difference for interaction between time periods and groups (F=1.912, P=0.003)., Conclusion: RNFL damage patterns are different under different IOP elevated courses. In APACG, RNFL was found to be swollen and thickening right after acute attack and then becomes thinning and atrophy along with the time, while RNFL was found to be diffused thinness in CPACG.

Objective: To evaluate the clinical function and significance of establishing a regional active neonatal transport network (ANTN) in Beijing., Method: The authors retrospectively studied intensive care and the role of ANTN system in management of critically ill neonates and compared the outcome of newborn infants transported to our NICU before and after we established standardized NICU and ANTN system (phase 1: July 2004 to June 2006 vs phase 2: July 2006 to May 2008)., Result: The number of neonatal transport significantly increased from 587 during phase 1 to 2797 during phase 2. Success rate of transport and the total cure rate in phase 2 were 97.85% and 91.99% respectively, which were significantly higher than those in phase 1 (94.36% and 88.69%, respectively, P < 0.01). The neonatal mortality significantly decreased in phase 2 compared with that in phase 1 (2.29% vs 4.31%, P < 0.01). The capacity of our NICU was enlarged following the development of ANTN. There are 200 beds for level 3 infants in phase 2, but there were only 20 beds in phase 1. Significantly less patients in the phase 2 had hypothermia, acidosis and the blood glucose instability than those in phase 1 (P < 0.01, 0.05, 0.01 and 0.05, respectively). The proportion of preterm infants transported to our NICU were higher in phase 2 compared with that in phase 1, especially infants whose gestational age was below 32 weeks. The proportions of asphyxia and respiratory distress syndrome were lower in phase 2 than that in phase 1, but the total cure rates of these two diseases had no significant changes between the two phases. The most important finding was that the improvement of outcome of premature infants and those with asphyxia and aspiration syndrome was noted following the development of ANTN., Conclusion: Establishing regional ANTN for a tertiary hospital is very important to elevate the total level in management of critically ill newborn infants. It plays a very important role in reducing mortality and improving total outcomes of newborn infants. There are still some problems remained to solve after four years practice in order to optimize the ANTN to meet needs of the development of neonatology.

Aim: To study preventive and therapeutic effect of anisodamine on acute respiratory distress syndrome(ARDS) induced by oleic acid and their mechanism of action., Methods: Model of ARDS was made in rabbits by oleic acid (OA). The effect of anisodamine on the malondialdehyde (MDA), fibronectin (FN), lactate dehydrogenase (LDH) and acid phosphatase (ACP) in plasma, and superoxide dismutase (SOD) in erythrocyte and MDA, SOD and pulmonary surfactant (PS) in lung tissues homogenate and pathological examination of lung were observed., Results: The administration of anisodamine before and after 30 minutes of injection OA decreased MDA, LDH and ACP, prevented the reduction of SOD, FN and PS. Compared with ARDS group, there was marked difference between the two, and alleviated lung injury., Conclusion: Anisodamine possesses preventive and therapeutic effects on ARDS by inhibiting lipid peroxidation and stabilizing membranes.

Uniform and vertical indium-oxide nanotube (IONT) arrays embedded well in n-type InP single crystal have been successfully prepared in situ by porous InP-template-assisted chemical vapor deposition (CVD). This IONT/InP nanostructure reveals high sensitivity to humidity at room temperature, which is ascribed to the ultrahigh surface-to-volume ratio of this nanostructure and the large number of oxygen defected states in IONTs. Such a nanostructure of IONT arrays embedded in a III-V semiconductor substrate could be expected to have potential applications, such as superior gas sensors. This work provides a novel approach for fabricating low-melting metal oxide semiconductor nanotubes.

Objective: To analyze the risk factors of retinopathy of prematurity (ROP) and provide evidence for the rational establishment of screening standard., Methods: The clinical data of 1675 preterm infants at gestational age < or = 36 weeks or birth weight < or = 2500 g who were admitted to the neonatal intensive care unit and had been screened in our hospital from July 2006 to May 2008 were analyzed retrospectively by univariate analysis and Logistic regression analysis. Gender, birth count, gestational age, birth weight, oxygen therapy, and mother's conditions were recorded., Results: ROP was detected in 195 (11.6%) of 1675 infants, of whom 35 infants (2.1%) had type 1 or threshold ROP. The lower the birth weight, the smaller the gestational age and the longer the time of oxygen therapy were, the higher the incidence of ROP was. For the infants whose birth weight was < or = 1200 g, 1201 - 1500 g, 1501 - 2000 g, 2001 - 2500 g, the incidence of ROP was 73.2%, 30.4%, 8.0%, and 1.1%; for those at gestational age < or = 30 weeks, 30(+1)-32 weeks, 32(+1)-34 weeks, 34(+1)-36 weeks, the incidence of ROP was 67.6%, 16.9%, 3.9%, and 1.0%; for the infants underwent oxygen therapy for 0 d, -3 d, -5 d, -8 d, > 8 d, the incidence of ROP was 1.5%, 3.3%, 9.6%, 23.2% and 38.8%;in the infants who inhaled oxygen at concentrations of 0.40, -0.60, -0.80 and > 0.80, the incidence of ROP was 11.8%, 18.1%, 26.8%, and 52.6%, respectively. Logistic regression analysis indicated that low birth weight, small gestational age, asphyxia, apnea, oxygen therapy were the high risk factors of ROP (the odds ratio was 0.957, 1.052, 1.186, 5.314, and 1.881)., Conclusions: Low birth weight, small gestational age, asphyxia, apnea, and oxygen therapy were the high risk factors of ROP. It is recommended that all preterm infants with high risk factors should be screened.

Aim: To investigate the effect of pretreatment with taurine on liver injury changes and the change of tumor necrosis factor alpha and NFkappaB expression following rats limbs ischemia/reperfusion., Methods: The model of limbs ischemia/reperfusion injury on rats was adopted in the experiment. Wistar rats were randomized into 4 groups (n = 10): Control group, T group, I/R group and TR group. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and MDA in the plasma, MDA, MPO, calcium in liver tissues were measured by colorimetric method. The content of TNF-alpha in plasma and liver tissues was determined by radioimmunoassay. The morphologic changes were observed with HE staining. The expressions of NF-kappaBp65 in liver tissues were tested by immuno-histochemistry method., Results: It was found that against the control group, the test values of ALT, AST, et al. and expressions of TNF-alpha, NF-kappaB increased in I/R group and TR group, but values of those in TR group were lower than in I/R group., Conclusion: Taurine can decrease the levels of TNF-alpha and NF-kappaB. It can mitigate the liver injury after limb ischemia/reperfusion injury in rats.

Background: The Sonic hedgehog (SHH) protein produced in the zone of polarising activity (ZPA) is a major determinant of the identity and numbers of digits in early limb development. Preaxial polydactyly types II (PPD2) and III (PPD3) have been mapped to a critical region at 7q36, and subsequently shown to be caused by point mutations in the ZPA regulatory sequence (ZRS), a long range cis-regulator for the SHH gene. Triphalangeal thumb-polysyndactyly syndrome (TPTPS) and syndactyly type IV (SD4) were also mapped to the 7q36 region but pathogenic mutations in ZRS have not yet been affirmed., Methods and Results: We performed linkage and haplotype analysis in six Han Chinese families with TPTPS and/or SD4, and refined the disease locus to an interval of 646 kb containing ZRS. In all families, the affected individuals heterozygous at rs10254391 (a single nucleotide polymorphism within ZRS) revealed a remarkable allele imbalance on sequence chromatogram. Using real-time quantitative polymerase chain reaction (qPCR), we identified duplication of ZRS and found that this duplication segregated with the limb phenotypes in all families but was not detected in unaffected family members or in unrelated control individuals. The duplication was also confirmed by interphase fluorescence in situ hybridisation (FISH) in an affected individual. We designed 17 additional qPCR assays and defined the minimum duplications in all six families, ranging from 131kb to 398kb., Conclusion: Both TPTPS and SD4 are due to duplications involving ZRS, the limb specific SHH enhancer. Point mutations in the ZRS and duplications encompassing the ZRS cause distinctive limb phenotypes.

To determine the survival and differentiation of cultured Human amniotic cells (HACs) upon transplantation into the brain of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson disease (PD) mice. Mouse model of PD was established with injections of MPTP (15 mg/kg, fourth, 2 h interval). After being labeled with PKH26, HACs isolated from human were transplanted into the striatum of PD mice. Immunohistochemistry was performed to evaluate the toxicity of MPTP in the substantia nigra, graft survival and endogenous neurogenesis. Brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) level in the striatum were tested by ELISA. Our results showed that cultured HACs can express the marker of neural progenitor cells and differentiate into neuron, dopaminergic neuron, astrocyte and oligodendrocyte. TH-positive neural cells were significantly reduced in the substantia nigra in the model mice, whereas which increased in transplantation mice. Immunohistology results showed that transplanted HACs survived and migrated in the brain of PD model mouse, though no morphological integration was observed. BrdU-positive cells in the Subventricular zone (SVZ) and neurotrophins of the striatum increased in the transplantation mice. The results suggested that transplanted HACs could survive and promote the endogenous neurogenesis of mice, which maybe related to the increased level of neurotrophins of the striatum.

Calcium-dependent protein kinases (CDPKs) are crucial sensors of calcium concentration changes in plant cells under diverse endogenous and environmental stimuli. We identified 20 CDPK genes from bread wheat and performed a comprehensive study on their structural, functional and evolutionary characteristics. Full-length cDNA sequences of 14 CDPKs were obtained using various approaches. Wheat CDPKs were found to be similar to their counterparts in rice in genomic structure, GC content, subcellular localization, and subgroup classification. Divergence time estimation of wheat CDPK gene pairs and wheat-rice orthologs suggested that most duplicated genes already existed in the common ancestor of wheat and rice. The number of CDPKs in diploid wheat genome was estimated to be at least 26, a number close to that in rice, Arabidopsis, and poplar. However, polymorphism among EST sequences uncovered transcripts of all three homoeologous alleles for 13 out of 20 CDPKs. Thus, the hexaploid wheat should have 2-3 fold more CDPK genes expressing in their cells than the diploid species. Wheat CDPK genes were found to respond to various biotic and abiotic stimuli, including cold, hydrogen peroxide (H(2)O(2)), salt, drought, powdery mildew (Blumeria graminis tritici, Bgt), as well as phytohormones abscisic acid (ABA) and gibberellic acid (GA). Each CDPK gene often responded to multiple treatments, suggesting that wheat CDPKs are converging points for multiple signal transduction pathways. The current work represents the first comprehensive study of CDPK genes in bread wheat and provides a foundation for further functional study of this important gene family in Triticeae.

Objective: To investigate the effect of Coptis chinensis on jaundice of G6PD deficient neonates., Method: 122 G6PD deficient neonates with jaundice who were in People' s Hospital of Guigang of Guangxi province from January 1999 to October 2004 were divided into two groups: C. chinensis group (62 neonates with C. chinensis administration before jaundice' s appearance) and none C. chinensis group (60 neonates without C. chinensis administration before jaundice' s appearance). The initial time, duration of jaundice, hemoglobin and serum bilirubin level and the incidence of kernicterus were analyzed between the two groups., Result: The initial time of jaundice is significantly later and the duration of jaundice is markedly shorter in the neonates with C. chinensis than that without C. chinensis. Simultaneously, the level of hemoglobin is significantly increased, and there is a low tendency of serum total bilirubin and direct bilirubin level in C. chinensis group as compared to that in none C. chinensis group. Moreover, there is no kernicterus in C. chinensis group and no difference in the treating result out of hospital between the two groups., Conclusion: Our results do not support the view that C. chinensis could aggravate jaundice of G6PD deficient neonates.

Objective: To investigate the association between Gln223Arg variant in leptin receptor (LEPR) gene and obesity., Methods: The subjects were from Yu county, Shanxi province, China. Data on cardiovascular risk factors was collected with questionnaire and their Gln223Arg variants in LEPR gene were genotyped. The relation between Gln223Arg variant in LEPR and obesity was analyzed., Results: The genotype frequencies of G. G, G. A and AA were 0.7679, 0.2171 and 0.015 respectively and the allele frequencies of G and A were 0.8764 and 0.1236. The distribution of genotype was in Hardy-Weinberg equilibrium (P = 0.934). The results of single factor analysis showed that the carriers with the Arg223 alleles had higher weight (63.2 kg vs. 61.9 kg; P= 0.0307) and body mass index (24.4 kg/rn vs. 24.1 kg/m2; P = 0.0898) than those noncarriers of these alleles. Results from multivariate analysis indicated that the carriers with the Arg223 alleles had higher body mass index (24.5 kg/m2 vs. 24.1 kg/m ;P= 0.0396) and prevalence of obesity (OR = 1.30, 95% CI: 0.957-1.767; P= 0.0935)., Conclusion: These data indicated that Gln223Arg variant in LEPR gene was possibly associated with obesity, respectively.

Objective: To investigate the expression patterns of surfactant protein B (SP-B) and its role in the development of human fatal lung epithelial cells., Methods: Human fetal lung tissues were obtained from 37 fetuses of 10-34 weeks at abortion with parental consent and from two newborn infants who died of non-pulmonary causes. SP-B expression in the lung tissues was examined by immunohistochemistry., Results: SP-B was detected in the cytoplasm of nonciliated columnar epithelial cells of the human fetal lung in as early as the 16th week of gestation. The positive reaction of SP-B was enhanced during canalicular stages and was more intense in the distal than in the proximal airway epithelium. From the 25th week to the prenatal stage, SP-B expression underwent no significant changes in the primitive alveolar stage, but increased remarkably after birth., Conclusion: The expression and secretion of SP-B reflects the maturation of the epithelial cells in human fatal lungs, and may closely associate with the survival ability of the newborn infants.

The use of full-length cDNA libraries is an effective tool to obtain complete gene information in a high-efficiency, high-throughput manner, especially in organisms with huge genomes that are not amenable to whole genome sequencing. In this review, we outlined several methods of full-length cDNA library construction and compared their advantages and disadvantages based on their respective principles. Drawing on our own experience, we described the Cap-trapper method in detail, with an emphasis on its application in wheat full-length cDNA library construction as well as the determination of the ratio of full-length cDNA in a library.

Objective: Recent progress in developmental biology has shown that the thyroid transcription factor-1 (TTF-1) plays an important role in lung development. The aim of this study was to investigate the expression and distribution of TTF-1 and its function during the development of epithelial stem cells in fetal human lungs., Methods: Human lung tissues were obtained with parental consent from 32 fetuses (10-27 weeks) and from seven newborn infants (28-36 weeks) who had not died from pulmonary diseases. The expression of TTF-1 was examined by immunohistochemistry., Results: TTF-1 was expressed in the nuclei of columnar nonciliated epithelial cells of the fetal human lung as early as 10 weeks of gestation. With the development of bronchus TTF-1 positive cells were present in scattered nonciliated cells and were predominantly expressed in the nuclei of epithelial cells of the distal tubules and lung buds. By the late phase of fetal development or neonatal period, TTF-1 was expressed in only type II alveolar epithelium cells and their precursor cells but was absent in ciliated cells and type I alveolar epithelium cells., Conclusions: TTF-1 can stimulate the growth of both bronchial trees and alveolar cells and regulate the type II alveolar epithelium cells and their precursors to secret surfactants.

Although the study of imprinted genes in human development is very important, little is known about their expression and regulation in the early differentiation of human tissues due to lack of an appropriate model. In this study, a Chinese human embryonic stem (hES) cell line, SHhES1, was derived and fully characterized. Expression profiles of human imprinted genes were determined by Affymetrix Oligo micro-array in undifferentiated SHhES1 cells and SHhES1-derived embryoid bodies (EBs) at day 3, 8, 13 and 18. Thirty-two known human imprinted genes were detected in undifferentiated ES cells. Significantly, differential expression was found in nine genes at different stages of EB formation. Expression profile changes were confirmed by quantitative real-time reverse transcriptase-polymerase chain reaction in SHhES1 cells as well as in another independently derived hES cell line, HUES-7. In addition, the monoallelic expressions of four imprinted genes were examined in three different passages of undifferentiated ES cells and EBs of both hES cell lines. The monoallelic expressions of imprinted genes, H19, PEG10, NDNL1 and KCNQ1 were maintained in both undifferentiated hES cells and derived EBs. More importantly, with the availability of maternal peripheral blood lymphocyte sample, we demonstrated that the maternal expression of KCNQ1 and the paternal expression of NDNL1 and PEG10 were maintained in SHhES1 cells. These data provide the first demonstration that the parental-specific expression of imprinted genes is stable in EBs after extensive differentiation, also indicating that in vitro fertilization protocol does not disrupt the parental monoallelic expression of the imprinted genes examined.

We established a genetic linkage map employing 518 simple sequence repeat (SSR, or microsatellite) markers for Bombyx mori (silkworm), the economically and culturally important lepidopteran insect, as part of an international genomics program. A survey of six representative silkworm strains using 2,500 (CA)n- and (CT)n-based SSR markers revealed 17-24% polymorphism, indicating a high degree of homozygosity resulting from a long history of inbreeding. Twenty-nine SSR linkage groups were established in well characterized Dazao and C108 strains based on genotyping of 189 backcross progeny derived from an F(1) male mated with a C108 female. The clustering was further focused to 28 groups by genotyping 22 backcross progeny derived from an F(1) female mated with a C108 male. This set of SSR linkage groups was further assigned to the 28 chromosomes (established linkage groups) of silkworm aided by visible mutations and cleaved amplified polymorphic sequence markers developed from previously mapped genes, cDNA sequences, and cloned random amplified polymorphic DNAs. By integrating a visible mutation p (plain, larval marking) and 29 well conserved genes of insects onto this SSR-based linkage map, a second generation consensus silkworm genetic map with a range of 7-40 markers per linkage group and a total map length of approximately 3431.9 cM was constructed and its high efficiency for genotyping and potential application for synteny studies of Lepidoptera and other insects was demonstrated.

Aim: To study the effects of PKC activation on apoptosis during ischemia/reperfusion in L-6TG rat skeletal myoblasts., Methods: Cultured L-6TG cells were divided into 3 groups: control group (C), ischemia/reperfusion group (I/R), PMA + ischemia/ reperfusion group (PMA), SOD, XOD and free calcium and mitochondrial respiration in L-6TG cell were evaluated in each group. Apoptosis was detected by flow cytometer with PI staining method and agarose gel electrophoresis, the immunohistochemical method was used to determine the expression of caspase-3., Results: Compared with I/R group, in PMA group, XOD , free calcium in L-6TG cell and apoptotic percentage all decreased significantly, while SOD and mitochondrial respiration in L-6TG cell increased. DNA fragmentation analysis of L-6TG cell showed no laddering pattern. The expression of caspase-3 was down regulated significantly., Conclusion: Activation of PKC can lessen ischemia/reperfusion injury and apoptosis through lessening oxidative injury and mitochondrial injury, adjusting calcium dyshomeostasis and down expression of caspase-3.

A powdery mildew resistance gene from Triticum urartu Tum. accession UR206 was successfully transferred into hexaploid wheat (Triticum aestivum L.) through crossing and backcrossing. The F1 plants, which had 28 chromosomes and an average of 5.32 bivalents and 17.36 univalents in meiotic pollen mother cells (PMC), were obtained through embryos rescued owing to shriveling of endosperm in hybrid seed of cross Chinese Spring (CS) x UR206. Hybrid seeds were produced through backcrossing F1 with common wheat parents. The derivative lines had normal chromosome numbers and powdery mildew resistance similar to the donor UR206, indicating that the powdery mildew resistance gene originating from T. urartu accession UR206 was successfully transferred and expressed in a hexaploid wheat background. Genetic analysis indicated that a single dominant gene controlled the powdery mildew resistance at the seedling stage. To map and tag the powdery mildew resistance gene, 143 F2 individuals derived from a cross UR206 x UR203 were used to construct a linkage map. The resistant gene was mapped on the chromosome 7AL based on the mapped microsatellite makers. The map spanned 52.1 cM and the order of these microsatellite loci agreed well with the established microsatellite map of chromosome arm 7AL. The resistance gene was flanked by the microsatellite loci Xwmc273 and Xpsp3003, with the genetic distances of 2.2 cM and 3.8 cM, respectively. On the basis of the origin and chromosomal location of the gene, it was temporarily designated PmU.

Aim: To observe the effect of phenolic alkaloids of Menispermum dauricum (PAMD) on thrombosis and platelet aggregation, and to explore its mechanism of action., Methods: Thrombosis was observed with arteriovenous shunt thrombus model in rat; platelet aggregation was determined by Born's method; ultrastructure of platelet was observed by transmission electron microscope; TXB2 or 6-keto-PGF1alpha levels were assessed by radioimmunoassay; and NO was determined by colorimetric method., Results: PAMD dose-dependently inhibited experimental thrombus formation, platelet aggregation induced by ADP, AA and THR in vivo and ultrastructure changes stimulated by THR; PAMD increased the generation of 6-keto-PGF1alpha in thoracic aortae and NO level in plasma; and had no influence on TXB2 release (P > 0.05)., Conclusion: PAMD inhibited thrombosis and platelet aggregation, and its mechanism might be due to the increase of PGI2 and NO level.

To discover new genes in a throughput manner,the cap-trapper method published previously was optimized for raising the efficiency in the construction of full-length cDNA library. Using the optimized protocol,we successfully constructed a full-length cDNA library of Aegilops speltoides,which contained 3.0 x 10(6) clones and more than 99% of plaques were recombinant phages. Sequence analysis results indicated that more than 89% of the clones were full-length.

One 134 bp fragment was amplified in anthers of male sterile and fertile wheat using one pair of degenerated primer designed based on the conserved domain of MS2 gene in Arabidopsis thaliana and Oryza sativa, and one 1604 bp male sterility gene homology sequence was extended by in silico cloning based on the 134 bp fragment. The amino acids encoded by the male sterility gene homology sequence include a 200 amino acid conserved domain of male sterility, and this sequence expressed only in wheat male fertile anthers; no expression in male sterile anthers, roots and leaves. This research demonstrated that the cloned male sterile homology sequence is specific to wheat anther development.

Objective: To construct a high performance open source software engine based on IBM SPLASH algorithm for later research on pattern discovery., Methods: Gpat, which is based on SPLASH algorithm, was developed by using open source software., Results: GNU Pattern (Gpat) software was developped, which efficiently implemented the core part of SPLASH algorithm. Full source code of Gpat was also available for other researchers to modify the program under the GNU license., Conclusion: Gpat is a successful implementation of SPLASH algorithm and can be used as a basic framework for later research on pattern recognition in biological sequences.

Purpose: To investigate IRF6 gene mutation in a van Der Woude syndrome (VWS) family in Henan province., Methods: PCR and DNA sequencing was employed to detect the mutation of IRF6.Secondary construction transformation analysis was performed using PIX-Protein Identification software., Results: A CGC>TGC(r.279c-->t) transversion of IRF6 was identified in condon 6, showing complete segregation with the disease phenotypes and was resulting in changes of the secondary constructure of IRF6., Conclusion: VWS is caused by mutations in IRF6 gene, and IRF6 is closely related to the development of lip, palate and tooth.

This is the first time to described the dissimilatory Fe(III) reducing characteristics of Shewanella cinica D14T. The effects of O2, light, temperature and pH on dissimilatory Fe(III) reduction were examined. The results suggested that the rate of Fe(III) reduction decreased with increasing Fe(III) concentration. Fe(III) reduction was partially inhibited by the presence of either O2 or light. The optimum temperature for Fe(III) reduction is 37 degrees C. At pH 6.0-10.0, strain D14T can reduce Fe(III). The soluble Fe(III) is more easy to be reduced than the insoluble one. Results of protein denaturants SDS and OGP suggest that the Fe(III) reduction activity of S. cinica is mostly localized to the soluble outer membrane fraction. The azo dye decolorization and Fe(III) reduction in strain D14T were enhanced in the presence of Fe(III) and dye.

Aims: Hereditary non-polyposis colorectal cancer, thyroid medullary carcinoma, breast/ovarian cancer and gastric cancer/breast cancer syndrome are encountered in surgery. Some gastric cancer/breast cancer syndrome may be the result of a CDH1 germline mutation. This is the first report of CDH1 germline mutations gastric cancer/breast cancer syndrome in Chinese patients., Methods: Peripheral blood from the proband, as well as, her first and second degree relatives was collected and CDH1 gene exon 1-16 mutations were screened. E-cadherin/beta-catenin proteins expression and histopathologic features were examined on gastric cancer/breast cancer tissues from the proband., Results: A C-->T nucleotide substitution at exon 13 (mRNA 2200 locus, Accession number NM-004360) was found. This was a transition from GCC-->GCT in DNA sequence (Ala154Ala). Diffuse-type gastric cancer and infiltrating ductal breast carcinoma were present. Both tumours preserved E-cadherin/beta-catenin expression immunohistochemically., Conclusions: Familial cancer syndrome with diffuse-type gastric cancer/breast cancer proband in Chinese has a propensity of early onset during lifespan. No truncating or splice-site CDH1 mutations had been identified in this family. A silent nucleotide variation in exon 13 of the CDH1 gene may contribute to some forms of cancer susceptibility.

Objective: To investigate the expression and distribution of telomerase reverse transcriptase (hTERT) and its significance during the development of epithelial stem cells in human fatal lungs., Methods: Human lung tissues were obtained at abortion from 37 fetuses 10-34 weeks of gestational age with parental consent, and the expression of hTERT in these tissues was examined by immunohistochemistry., Results: In fetuses of 10 weeks old, hTERT was detected mainly in the epithelial cells of the proximal bronchi in the fetal lung, and migrated gradually to the distal bronchi as the lung developed. The hTERT expression level peaked at 17-20 weeks of gestation and then decreased over the period of alveolarization that occurred at about 25-26 weeks. In the later gestation period, strongly positive epithelial cells became concentrated in discrete patches near the basal membrane of the airway and scattered in only a few of the pulmonary alveoli. Morphologically, these cells resembled the basal cells and type II alveolar epithelial cells (AEC) or their progenitor cells., Conclusions: Telomerase expression and activity are hallmarks of the pulmonary stem or progenitor cells that maintain undifferentiated state and self-renewal capacity, and it plays a crucial role in the normal differentiation and regeneration of the bronchial and alveolar epithelial cells to maintain the epithelial integrity.

Freestanding single-crystal complete nanorings of zinc oxide were formed via a spontaneous self-coiling process during the growth of polar nanobelts. The nanoring appeared to be initiated by circular folding of a nanobelt, caused by long-range electrostatic interaction. Coaxial and uniradial loop-by-loop winding of the nanobelt formed a complete ring. Short-range chemical bonding among the loops resulted in a single-crystal structure. The self-coiling is likely to be driven by minimizing the energy contributed by polar charges, surface area, and elastic deformation. Zinc oxide nanorings formed by self-coiling of nanobelts may be useful for investigating polar surface-induced growth processes, fundamental physics phenomena, and nanoscale devices.

Objective: To observe the development of the alveoli and the differentiation of the epithelium in the fetal lung of Chinese., Methods: Chinese human lung tissues were obtained from 20 fetuses of 10 to 34 weeks old, and the ultrastructural changes of the alveolar epithelial cells were observed using both optical and transmission electron microscope., Results: At the first 10 to 16 weeks of development, the lung showed a tubular structure, the epithelial cells were columnar and poorly differentiated. At weeks 17 to 24, the bronchioles increased and the epithelial cells were differentiated to approach maturity. A few lamellar bodies were observed in the cuboidal lining cells with the majority being type II alveolar cells at 25 weeks, when the primitive alveolar was formed. In this stage many primitive epithelial cells could be seen, which appeared primordial in form before evolving into type II and type I cells and perhaps served as epithelial stem cells or progenitor cells. From about 26 weeks, type I cells were observed but most of the epithelial cells were type II cells. Type I cells began to increase in the alveolar epithelium until 32 weeks., Conclusions: The alveolar type II cells and type I cells are probably both derived from the same precursor cells or stem cells, and no evidence has shown that type II cells may differentiate into type I cells during the development of the fetal lung.

Two overlapping bacterial artificial chromosome (BAC) clones from the B genome of the tetraploid wheat Triticum turgidum were identified, each of which contains one of the two high-molecular-weight (HMW) glutenin genes, comprising the complex Glu-B1 locus. The complete sequence (285 506 bp of DNA) of this chromosomal region was determined. The two paralogous x-type ( Glu-1-1 ) and y-type ( Glu-1-2 ) HMW-glutenin genes of the complex Glu-B1 locus were found to be separated by ca. 168 000 bp instead of the 51 000 bp separation previously reported for the orthologous Glu-D1 locus of Aegilops tauschii, the D-genome donor of hexaploid wheat. This difference in intergene spacing is due almost entirely to be the insertion of clusters of nested retrotransposons. Otherwise, the orientation and order of the HMW glutenins and adjacent genes were identical in the two genomes. A comparison of these orthologous regions indicates modes and patterns of sequence divergence, with implications for the overall Triticeae genome structure and evolution. A duplicate globulin gene, found 5' of each HMW-glutenin gene, assists to tentatively define the original duplication event leading to the paralogous x- and y-type HMW-glutenin genes. The intergenic regions of the two loci are composed of different patterns and classes of retrotransposons, indicating that insertion times of these retroelements were after the divergence of the two wheat genomes. In addition, a putative receptor kinase gene near the y-type HMW-glutenin gene at the Glu-B1 locus is likely active as it matches recently reported ESTs from germinating barley endosperm. The presence of four genes represented only in the Triticeae endosperm ESTs suggests an endosperm-specific chromosome domain.

We report that the Zn-terminated ZnO (0001) polar surface is chemically active and the oxygen-terminated (000(-)1) polar surface is inert in the growth of nanocantilever arrays. Longer and wider "comblike" nanocantilever arrays are grown from the (0001)-Zn surface, which is suggested to be a self-catalyzed process due to the enrichment of Zn at the growth front. The chemically inactive (0001;)-O surface typically does not initiate any growth, but controlling experimental conditions could lead to the growth of shorter and narrower nanocantilevers from the intersections between (000(-)1)-O with +/- (01(-)10) surfaces.

Background: Porokeratosis palmaris et plantaris disseminata (PPPD) is a rare autosomal dominant dyskeratotic disorder characterized by a cornoid lamella with parakeratosis, hyperkeratosis and loss of granular layers. The genetic basis of this disease is still unknown. Two loci for disseminated superficial actinic porokeratosis (DSAP) were found to be located on 12q23.2-24.1 and 15q25.1-26.1. Both PPPD and DSAP are disseminated types of porokeratosis., Objectives: To locate the locus for PPPD, thereby facilitating the identification of this disease gene and leading to an understanding of the pathogenesis of porokeratosis., Methods: Genotyping was performed in a Chinese family with PPPD using polymorphic microsatellite markers on 12q and 15q., Results: The locus for PPPD is located within a 6.9-cM region between markers D12S1613 and D12S1341, with a maximum two-point LOD score of 8.14 (theta = 0.00) at D12S1335., Conclusions: This study provides a map location for isolation of a gene causing PPPD.

To solve the problem of immune incompatibility, nuclear transplantation has been envisaged as a means to produce cells or tissues for human autologous transplantation. Here we have derived embryonic stem cells by the transfer of human somatic nuclei into rabbit oocytes. The number of blastocysts that developed from the fused nuclear transfer was comparable among nuclear donors at ages of 5, 42, 52 and 60 years, and nuclear transfer (NT) embryonic stem cells (ntES cells) were subsequently derived from each of the four age groups. These results suggest that human somatic nuclei can form ntES cells independent of the age of the donor. The derived ntES cells are human based on karyotype, isogenicity, in situ hybridization, PCR and immunocytochemistry with probes that distinguish between the various species. The ntES cells maintain the capability of sustained growth in an undifferentiated state, and form embryoid bodies, which, on further induction, give rise to cell types such as neuron and muscle, as well as mixed cell populations that express markers representative of all three germ layers. Thus, ntES cells derived from human somatic cells by NT to rabbit eggs retain phenotypes similar to those of conventional human ES cells, including the ability to undergo multilineage cellular differentiation.

Aim: Recently, more SARS-CoV virus genome sequences are released to the GenBank database. The aim of this study is to reveal the evolution forces of SARS-CoV virus by analyzing the nucleotide mutations in these sequences., Methods: We obtained 20 SARS-CoV virus genome sequences from NCBI database, and calculated the ratio of non-synonymous nucleotide substitution per non-synonymous site (Ka) and synonymous nucleotide substitution per synonymous site (Ks) for SARS-CoV virus genes., Results: The Ka/Ks ratios for replicase polyprotein ORF1a, ORF1b, and spike protein gene are 1.09 (P=0.6501), 0.38 (P=0.0074), 0.65 (P=0.0685) respectively., Conclusion: SARS-CoV virus replicase polyprotein ORF1b is undergoing negative selection; negative selection force is also probably operating on spike protein gene. These results provide basis for future developing a new drug and vaccine against SARS.

More than a hundred of loci (genes) affect the development of mouse and human hair. A locus of Uncv (uncovered) has been confirmed to be involved in hairlessness for homozygote and sparse hair for heterozygote. Except hairlessness(or called uncovered coat), the homozygote was also accompanied by growth retard and puberty delay. Identification of the mutation in the gene will be important for understanding the related diseases in human. Although the uncovered locus (Uncv) has been mapped to the mouse distal chromosome 11(Chr11), the high-resolution genetic map and physical map of the locus has not been created. In this study, 2074 F2 mouse populations from backcross [BALB/c (Uncv/Uncv) x C3H (+/+)]x BALB/c (Uncv/Uncv) and [BALB/c (Uncv/Uncv) x C57BL/6 (+/+)] x BALB/c(Uncv/ Uncv) were genotyped using 16 polymorphic markers with an approximately 20 cM interval on mouse distal Chr11. By genetic linkage analysis, Uncv locus was mapped to an approximately 1.4 cM interval between markers D11Mit337 and D11Mit338 with the following order: proximal D11Mit338-D11Mit203 (Uncv)-D11Mit103 -D11Mit337 distal on mouse Chr11. And then, a contig of 35 BACs representing the Uncv-containing region was constructed. The contig covered 800-1000 kb region flanked by 189K10-SP6 and D11Mit103. Together, we have constructed the high-resolution genetic map and detailed physical map of the Uncv region. This will facilitate the identification of the Uncv loci.

This research was performed to investigate the relationship between 16 candidate genes responsible for dopaminergic transmission or iron metabolism and familial restless legs syndrome. Genotyping was performed in a Han restless legs syndrome family using the technique of fluorescence-based genescan with the microsatellite markers selected in chromosomal regions flanking the candidate genes. Classical linkage analysis was conducted under the autosomal dominant genetic mode. Results showed that all of the LOD scores at recombination fraction 0.00 are smaller than -2.00, which indicated that these loci were not linked to familial restless legs syndrome. No linkage was found between the candidate genes and RLS in this family. Familial restless legs syndrome may be caused by another gene related to dopaminergic transmission and iron metabolism or there is new mechanism involved in this disease.

Bacterial artificial chromosome (BAC) library plays a pivotal role in genomics studies. A crucial step in BAC library construction is preparation of BAC vector DNA. Preparation of highly purified vector DNA is affected by a series of factors including digestion of restriction enzyme and dephosphorylation of linearized vector DNA. In our study, the BAC vector pECBAC1 was digested by the restriction enzyme of BamHI and dephosphorylated by HK phosphatase respectively. In order to improve the ligation capability of vector DNA, gel purification of linearized vector DNA was also conducted in our study. At the same time, we did a series of experiments to get high quality of vector DNA for construction of BAC library. They included the optimal concentration of restriction enzyme, optimal digestion time, the type of phosphatase and gel purification of linearized vector DNA.

Fifteen organophosphorus pesticide residues in vegetable samples were extracted and cleaned-up by gel permeation chromatography and detected by capillary GC-FPD. An alternative validation method was established based on GC-NPD detection on an HP-1701 column. The eluting profiles of 15 compounds on a gel permeation column (10 mm i.d. x 200 mm, SX-3) eluted by 1 mL/min cyclohexane-ethyl acetate (volume ratio of 1:1) were established according to GC quantitation. Minimum detection limits of this method were between 5 x 10(-4) mg/kg to 5 x 10(-3) mg/kg. The average fortified recoveries and coefficients of variation of the method were determined. The results show that recoveries are between 77.8%-106%, with an average RSD of 0.10%-16.6%.

Objective: To study the DNA synthesis in the airway cells of asthmatic rats after allergen stimulation in association with airway remodeling., Methods: Double staining immunohistochemical techniques was used to determine DNA synthesis of the airway cells of 12 asthmatic and 12 normal rats. BrdU incorporation into the airway smooth muscle (ASM) and epithelium was quantified by employment of computer-assisted image analysis., Results: BrdU indices in both the ASM and the epithelium of asthmatic model group were higher than those of the control group (P<0.01, P<0.05), and positive linear correlation of the BrdU indices in the ASM and epithelium with the airway diameter was observed (r=0.7828, P<0.01; r=0.5852, P<0.05), which was not found in the control group (r=-0.3755, P>0.05; r=-0.5208, P>0.05). The epithelial thickness of the model group was significantly greater than that of the control group (P<0.01). There was no significant difference in terms of airway diameter, thickness of the ASM and the area positive of alpha-smooth muscle actin between the 2 groups (P>0.05)., Conclusion: Increased DNA synthesis and accelerated proliferation of ASM and epithelial cells in sensitized SD rats following repeated allergen challenges may lead to airway remodeling.

A wheat line, Bai Nong 3217/Mardler BC5F4 with resistance to powdery mildew, was used to construct a conventional cDNA library and a suppression subtractive hybridization (SSH) cDNA library from wheat leaves inoculated by Erysiphe graminis DC. Three hundred and eighty-seven non-redundant ESTs from the conventional cDNA library and 760 ESTs from the SSH cDNA library were obtained, and the ESTs similarity analysis using BLASTn and BLASTx were conducted by comparing these ESTs with sequences in GenBank. The results showed that the redundancy of some kinds of genes such as photosynthesis related genes and ribosome related genes was higher in the conventional cDNA library but the varieties and quantities of disease resistance genes were less than those in the SSH cDNA library. The SSH cDNA library was found to have obvious advantages in gene expression profiling of disease resistance such as simple library construction procedure, rich specific DRR (disease-resistance-related) genes and decreased sequencing amount. To acquire genes that were involved in the powdery mildew resistance of wheat, hybridization with high-density dots membranes was used to screen the two libraries. The result showed that the method was relatively simple in operation, and the membranes could be used for many times. But some problems also existed with this screening method. For instance, a large amount of mRNA and radioactive isotope were needed and the hybridization procedure must be repeated several times to obtain stable hybridization results. About 54.1% function-known ESTs in the SSH cDNA library were identified to be DRR genes by screening. There were 247 clones of the SSH cDNA library that had positive signal in the repeated hybridizations with the pathogen uninfected probe. The identified DRR genes distributed in the whole procedure of powdery mildew resistance, but mainly focused on the SAR (systemic of acquired resistance).