Marker-assisted breeding for transgressive seed protein content in soybean [Glycine max (L.) Merr].

KEY MESSAGE: After two cycles of marker-assisted breeding on three loci, lines with transgressive segregation of 8.22-9.32 % protein content were developed based on four original soybean parents with 35.35-44.83 % protein content. Marker-assisted breeding has been an innovative approach in conventional breeding, which is to be further demonstrated, especially for quantitative traits. A study on continuous transgressive breeding for seed protein content (SPC) in soybean using marker-assisted procedures is reported here. The SPC of the recombinant inbred line (RIL) population XG varied in 38.04-47.54 % under five environments with P 1 of 35.35 %, P 2 of 44.34 % and total heritability of 89.11 %. A transgressive segregant XG30 with SPC 45.53 % was selected for further improvement. The linkage mapping of XG showed its genetic constitution composed of five additive QTL (32.16 % of phenotypic variation or PV) and two pairs of epistatic QTL (2.96 % PV) using 400 SSR markers with the remnant heritability 53.99 % attributed to the undetected collective of minor QTL. Another transgressive segregant WT133 with SPC 48.39 % was selected from the RIL population WT (44.83 % SPC for both parents). XG30 and WT133 were genotyped on the three major additive QTL (Prot-08-1, Prot-14-1 and Prot-19-2) as A 2 A 2 B 2 B 2 L 1 L 1 and A 1 A 1 B 1 B 1 L 2 L 2 , respectively. From WT133×XG30, surprising transgressive progenies were obtained, among which the recombinants with all three positive alleles A 2 _B 2 _L 2 _ performed the highest SPC, especially that of Prot-08-1. The five F 2-derived superior families showed their means higher than the high parent value in F 2:3 and F 2:4 and more transgressive effect in F 2:5:6, with the highest as high as 54.15 %, or 4.82 and 9.32 % more than WT133 and its original high parent, respectively. This study demonstrated the efficiency of marker-assisted procedure in breeding for transgressive segregation of quantitative trait.

Genome-wide identification and transcription analysis of soybean carotenoid oxygenase genes during abiotic stress treatments.

Carotenoid oxygenase is a key enzyme in carotenoid metabolism leading to the synthesis of two phytohormones, abscisic acid (ABA) and strigolactone, as well as norisoprenoids. Few studies have analyzed inter-relationship of the metabolic networks of these three substances. In this present paper, soybean carotenoid oxygenase genes were identified to reveal their phylogenetic relationships, and the transcriptional response of these genes to four abiotic stresses (NaCl, PEG, high and low temperature) and ABA treatment were investigated to characterize their potential roles in plant resistance. Positive selection was found in the branches of carotenoid cleavage dioxygenase (CCD1), CCD8 and NCED (9-cis-epoxycarotenoid oxygenase), indicating an adaptive evolution in these clades. In soybean eight carotenoid oxygenase genes were identified. The transcriptional responses of almost all of them under stress and ABA conditions were significantly altered when assessed by quantitative polymerase chain reaction. Notably, CCD1 and CCD4, previously known as the key genes in norisoprenoids metabolism, showed especially strong responses to the abiotic stresses and ABA treatment. Furthermore, transcription levels of CCD7 and CCD8, key genes for the strigolactone pathway, highly increased during ABA treatment providing further evidence that ABA is involved in regulating strigolactone metabolism. All of the carotenoid oxygenase genes in soybean are involved in plant abiotic stress physiology, and ABA is presumed to be a core regulatory substance. These findings provide some insights into the mechanisms that underlie the regulation of tolerance response to abiotic stresses in soybean.

[The characteristics and clinical manifestation of subjects with non-specific pattern of pulmonary function tests].

OBJECTIVE: To analyze the characteristics of pulmonary function and the clinical significance of non-specific pattern (NSP). METHODS: A total of 1933 pulmonary function tests of adult patients were analyzed, and those with NSP were selected. The pulmonary function test results, clinical diagnosis and radiological manifestations were analyzed. Normal distribution data were compared by t test, while non-normal distribution data were compared by Mann-Whitney U test, and χ(2) test was used to compare ratios. RESULTS: There were 61 patients in the NSP group and 1017 in the control group. The BMI of the 2 groups was (24.5 ± 4.6) and (24.5 ± 3.8) kg/m(2), respectively, being not significantly different (t = 0.008, P > 0.05). The age was 64 (49-74) years and 56 (42-70) years, and the smoking index was 0.4 (0-20) and 0 (0-10), respectively, showing no significant differences (Z values were -2.209 and -2.571, respectively, all P < 0.05). In the NSP group, FEV1 was 69% (66%-73%) predicted, FVC 75% (70%-77%) predicted, FEV1/FVC 75% (73%-78%), RV 118% (105%-145%) predicted, and TLC 86% (82%- 93%) predicted, which were significantly different as compared to those of the control group [95% (87%-104%), 98% (90%-106%), 79% (76%-84%), 101% (88%-114%) and 94% (88%-102%), respectively],(Z values are -13.059--5.185, all P < 0.05). RV/TLC was (52 ± 11)% in the NSP group and (39 ± 9)% in the control group, the difference being significant (t = -10.351, P < 0.05). The decreased TLC indicated restricted ventilation, while the increased RV and RV/TLC indicated air trapping. The clinical diagnosis of NSP included obstructive and restrictive diseases, some of which showed severe radiological abnormalities, but there were 31 patients without pulmonary lesions. CONCLUSIONS: Age and smoking, but not obesity, may play a role in NSP.NSP has characteristics of obstructive and restrictive ventilation defects, but does not associate with particular diseases, thus having limited clinical significance.

[Interventional effects of different management programs on the outcomes of stable chronic obstructive pulmonary disease patients].

OBJECTIVE: To evaluate the interventional effects of different management programs on the outcomes of stable chronic obstructive pulmonary disease (COPD) patients. METHODS: Systemic education, follow-up and control groups were divided according to the frequency of follow-ups and the profile of participating in education. A total of 157 patients were enrolled into the COPD database from May 2002 to May 2010. They were interviewed face-to-face at our department. The investigation contained general conditions, the frequency of acute exacerbations (AE) the previous year, COPD Assessment Test (CAT), Modified British Medical Research Council Dyspnea Scale (mMRC) and spirometric classification. A combined assessment was conducted. RESULTS: The frequency of AE and rate of AE < once the previous year in systemic education group was 0.9 ± 1.1 and 71.2%, 1.0 ± 0.8 and 68.6% in follow-up group and 1.4 ± 1.1 and 44.4% in control group. And the frequency of AE in systemic education and follow-up groups was significantly less than that in control group (P < 0.05). Rate of AE < once in systemic education and follow-up groups was significantly higher than that in control group (P < 0.01). CAT and rate of CAT ≤ 20 in systemic education, follow-up and control groups were 10.0 ± 5.0 and 96.2%, 11.1 ± 6.0 and 88.2%, 15.3 ± 6.8 and 64.8% respectively. CAT in systemic education and follow-up groups was significantly lower than that in control group (P < 0.01). Rate of CAT ≤ 20 in systemic education and follow-up groups was significantly higher than that in control group (P < 0.01). mMRC in systemic education, follow-up and control groups was 1.5 ± 0.8, 1.6 ± 0.9 and 2.1 ± 1.0 respectively. mMRC in systemic education and follow-up groups was significantly lower than that in control group (P < 0.05). Combined assessment showed that no significant difference existed in Types A and B among three groups (P > 0.05). Type C in the systemic education group was significantly higher than that of the control (P < 0.05). Type D in the systemic education and follow-up groups was significantly lower than that of the control (P < 0.01). CONCLUSION: Long-term systemic education and follow-up management program can reduce the frequency of AE of COPD effectively and improve the health status of COPD patients.

A novel nomogram and risk classification system for predicting lymph node metastasis of breast mucinous carcinoma: A SEER-based study.

BACKGROUND: Mucinous breast cancer (MBC) is a rare disease, and patients with lymph node metastasis (LNM) have a poor prognosis. We aimed to explore the predictive factors of LNM and to construct a nomogram for predicting the risk of LNM and to identify the suitable axillary surgery for patients with diverse risks. PATIENTS AND METHODS: Data were extracted from the Surveillance, Epidemiology, and End Results (SEER) database. Chi-square and rank-sum tests were used to analyze the differences between groups. Survival analysis was performed with Kaplan-Meier curves and log-rank tests. Independent factor identification and nomogram construction were performed with logistic regression analysis. The nomogram was qualified with a discrimination and calibration plot. Propensity score matching was performed to balance the disparities between groups. RESULTS: Patients with metastatic lymph nodes have a worse prognosis. Univariate and multivariate analyses indicated that tumor size, grade, and age were independent risk factors for LNM. The nomogram constructed with these three factors can predict the risk of LNM with high accuracy (AUC: 0.767, 95% CI: 0.697-0.838) and good calibration. Based on the nomogram, a risk classification system satisfactorily stratified the patients into 3 groups with diverse risks of LNM. In the low-risk group, there were no significant differences between sentinel lymph node biopsy and no axillary surgery. In the middle- and high-risk groups, both SLNB and axillary lymph node dissection were superior to no axillary surgery, with similar survival benefits. CONCLUSIONS: The nomogram based on tumor size, grade, and age could conveniently and accurately predict the risk of LNM in MBC and assist clinicians in optimizing surgical strategies.

[Transgenic technology and soybean quality improvement].

Soybean is an important source of edible oil, protein and protein diet. The breeding process of high quality soybean can be accelerated via employment of transgenic technology, by which the key genes for soybean quality traits could be directly manipulated. Thus, various soybean varieties could be bred to fulfill different needs for specific consumers. Here, we reviewed the contribution of transgenic technology to improvement of soybean qualities in recent years. We also introduce some newly developed safe transgenic technologies and hope this information could relieve some concerns on the GM food.

Global Transcriptome Profiling of Enterobacter Strain NRS-1 in Response to Hydrogen Peroxide Stress Treatment.

Microbes are often subjected to oxidative stress in nature that badly affects their growth rate and viability. Although the response of microbes against oxidative stress has been characterized at the chemical, physiological, and molecular levels, the mechanism of gene-regulation network adaptations of bacteria in response to oxidative stress remains largely unknown. In this study, transcriptomic profiling of glyphosate-tolerant Enterobacter strain NRS-1 was analyzed under 9 mM H2O2 stress using RNA-seq and qRT-PCR. The lag period in the growth of NRS-1 was very short compared with wild-type strain under H2O2 treatment. A total of 113 genes are identified as differentially expressed genes (DEGs) under H2O2 that include 38 upregulated and 75 downregulated transcripts. But not any genes regulated by major oxidative regulons, viz., oxyR, soxR, rpoS, perR, ohrR, and σв, have been reported in DEGs, hence potentially reflecting that specific changes have occurred in NRS-1 for adaptation to oxidative stress. Based on the functions of the DEGs, six elements namely formate dehydrogenase, processes associated with iron ions, repair programs, multidrug resistance, antioxidant defense, and energy generation (mqo, sdhC) might have contributed for stress tolerance in NRS-1. These elements are proposed to form a molecular network explaining gene response of NRS-1 to stress, and ensure global cell protection and growth recovery of NRS-1. These findings enrich the view of gene regulation in bacteria in response to H2O2 oxidative stress.

Complex gene response of herbicide-resistant Enterobacter strain NRS-1 under different glyphosate stresses.

Knowledge of biological evolution and genetic mechanisms is gained by studying the adaptation of bacteria to survive in adverse environmental conditions. In this regard, transcriptomic profiling of a glyphosate-tolerant Enterobacter strain NRS-1 was studied under four different treatments to investigate the gene-regulatory system for glyphosate tolerance. A total of 83, 83, 60 and 74 genes were up-regulated and 108, 87, 178 and 117 genes down-regulated under 60-NPG, 110-NPG, NaCl (355 mM) and HCl (pH 4.46) stress treatments, respectively. Complex gene network was identified to be involved in regulating tolerance to glyphosate. This study revealed that NRS-1 has gained glyphosate tolerance at the cost of osmotic and acidic resistance. The 25 differentially expressed genes are reported to may have partly changed the function for providing resistance to glyphosate directly, among them genes metK, mtbK, fdnG and wzb that might detoxify/degrade the glyphosate. However, under 110-NPG condition, NRS-1 might have utilized economical and efficient ways by depressing its metabolism and activity to pass through this stress. Hence, the present study provides insights into the genes involved in glyphosate tolerance, which can be effectively utilized to engineer herbicide-resistant crop varieties after their proper validation to manage weed growth.

Expression of CD74 in bladder cancer and its suppression in association with cancer proliferation, invasion and angiogenesis in HT-1376 cells.

The aim of the present study was to investigate the expression and potential roles of CD74 in human urothelial cell carcinoma of the bladder (UCB) in vitro and in vivo. CD74 and macrophage migration inhibitory factor (MIF) were located and assayed in normal and UCB samples and cell lines using immunostaining. CD74 was knocked down using CD74 shRNA lentiviral particles in HT-1376 cells. The proliferative, invasive potential and microvessel density (MVD) of knockdown-CD74 HT-1376 cells were analyzed in vitro or in vivo. The expression of CD74 in an additional high grade UCB J82 cell line was also verified in vivo. All experiments were repeated at least 3 times. The majority of muscle-invasive bladder cancer (MIBC) samples, and only one high grade UCB cell line, HT-1376, expressed CD74, compared with normal, non-muscle-invasive bladder cancer (NMIBC) samples and other cell lines. The levels of proliferation and invasion were decreased in the CD74 knockdown-HT-1376 cells, and western blotting assay indicated that the levels of proteins associated with proliferation, apoptosis and invasion in the cells were affected correspondingly by different treatments in vitro. The tumorigenesis and MVD assays indicated less proliferation and angiogenesis in the knockdown-HT-1376 cells compared with the scramble cells. Notably, J82 cells exhibiting no signal of CD74 in vitro presented the expression of CD74 in vivo. The present study revealed the potential roles of CD74 in the proliferation, invasion and angiogenesis of MIBC, and that it may serve as a potential therapeutic target for UCB, but additional studies are required.

MAN1B1 is associated with poor prognosis and modulates proliferation and apoptosis in bladder cancer.

Bladder cancer (BC) has been regarded as the most common malignancy of the urinary system worldwide. With lack of investigations for molecular pathogenesis underlying that develop BC, the therapeutic efficacy of several therapeutic approaches existing is still unsatisfactory. Here, our study aimed to explore the potentially biological function of MAN1B1 on BC. In this study, MAN1B1 expression level in BC tissues and normal tissues was analyzed based on The Cancer Genome Atlas (TCGA) data and correlation between its expression and prognosis was determined using Kaplan-Meier analysis. Knockout of MAN1B1 was performed using silencing RNA and the efficacy of MAN1B1 knockout was identified using quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. The BC cells proliferation was assessed by Cell Counting Kit-8 (CCK8) assay, and then the cells apoptosis was detected by Annexin V-fluorescein isothiocyanate (Annexin V-FITC)/propidium iodide (PI) staining and flow cytometry following MAN1B1 knocked down by small interfering RNA. Protein kinase B (AKT) signaling was evaluated by detecting related markers, namely AKT, p-AKT, 4E-BP-1 and Bax using western blot assay. As a result, the MAN1B1 expression was higher in BC tissues than those in normal tissues, besides, its overexpression was associated with poor prognosis. Moreover, MAN1B1 reduction by silencing RNA approach resulted in BC cells proliferation suppression and BC cells apoptosis promotion. Finally, AKT signaling activity was inhibited by MAN1B1 silencing. Taken together, these results unraveled that MAN1B1 may act on an oncogenic action in BC, which improved the likelihood of MAN1B1 taking on a promising prognostic biomarker and a potential target for treating BC.

[Mapping QTLs related to oil content of soybeans].

The soybean Recombinant Inbred Lines(RIL), including 133 lines, from the cross Wan82-178 x Tongshan-baopihuangdoujia were used as experimental materials in this study. Based on the linkage map constructed with Single Sequence Repeat(SSR) markers using this RIL population, the software Cartgrapher(V.2.0) and the composite interval mapping were employed to identify quantitative traits loci(QTL) associated with oil content of soybean in 2004 and 2005. It was found that the results of mapping QTL for the oil content were similar for these two years. They were both mapped near satt331 on linkage group wt-11, and they could be used to explain 13.95% and 15.01% of the total variation of the oil content, respectively. In addition, the software QTL Mapper 1.6 was applied to detect QTLs related to oil con-tent in two years. The result indicated that the QTL related to oil content was still mapped near satt331 on linkage group wt-11.

[Mapping insect resistance QTLs of soybean with RIL population].

A recombinant inbred line (RIL) population, NJRIKY, which was derived from the cross Kefeng 1 xNannong 1138-2, was used to constructed the genetic linkage map. Larval weight and pupae weight of cotton worm [Prodenia litura (L.) Fabricius] were examined and used as indicators of resistance. Based on the linkage map constructed with SSR markers of this RIL population, one QTL responsible for larval weight was mapped on linkage group G20-O and the position was 31.91 cM. The QTL's additive effect was 0.0408 and explained 11.74 of the total variation of the larval weight. Two QTLs associated with pupae weight were mapped on linkage group G8-D1b+W and G17-L and the positions were 14.71 cM and 0.01 cM, respectively. The QTLs' additive effects were -0.0139 and 0.0103 ,which explained 11.30 and 6.36 of the total variation of larval weight, respectively.

[The effect of chronic intermittent hypoxia caused by obstructive sleep apnea hypopnea syndrome on blood pressure].

OBJECTIVE: To explore the effect of chronic intermittent hypoxia caused by obstructive sleep apnea hypopnea syndrome (OSAHS) and chronic continuous hypoxia caused by chronic obstructive pulmonary disease (COPD) on blood pressure and levels of nitric oxide (NO)/endothelin (ET). METHODS: A total of 85 cases were selected, including OSAHS patients over 18 years old visited this hospital from June to August 2006, stable COPD patients and healthy volunteers. According to the results of clinical questionnaire, pulmonary function test and polysomography (PSG), they were divided into 4 groups: non-smoking OSAHS patients (n = 26), smoking OSAHS patients (n = 22), patients with stable COPD (n = 17) and healthy control subjects (n = 20). Blood pressure measurement was performed before and after the PSG examination while the subjects were resting. The levels of ET and NO in blood samples and exhaled breath condensates (EBC) were measured by radioimmunoassay and nitrate reductase, respectively. Measurement data were analyzed by ANOVA, numeration data were analyzed by chi-square test, data of normal distribution were analyzed by Pearson correlation analysis, and non-normal data were analyzed by Spearman correlation analysis. Multiple linear regression analysis was also performed. RESULTS: Regardless of the smoking status, the difference of morning-evening diastolic blood pressure (DBP) in the morning of patients with OSAHS [non-smoking OSAHS: (88 +/- 10) mm Hg, 1 mm Hg = 0.133 kPa; smoking OSAHS: (95 +/- 17) mm Hg] was higher than that of patients with COPD [(76 +/- 7) mm Hg] and healthy subjects [(70 +/- 6) mm Hg]. The difference of morning-evening NO level in EBC of patients with OSAHS [non-smoking OSAHS: (-4.5 +/- 7.9) micromol/L; smoking OSAHS: (-3.4 +/- 5.5) micromol/L] was lower than that of patients with COPD [(1.4 +/- 6.1) micromol/L] and healthy subjects [(3.1 +/- 4.0) micromol/L]. The ratio of NO to ET in serum of the smoking OSAHS group (1.0 +/- 0.5) was lower than that of the COPD group (1.4 +/- 0.7). After adjustment for sex, age, BMI, waist circumference, history of smoking and alcohol, level of hypertension, and category of antihypertensive drugs, the morning-evening difference of NO level in EBC was negatively associated with the morning-evening difference of DBP (r = -0.301, P = 0.021), and was positively associated with apnea hypopnea index (AHI) (r = 0.116, P < 0.05). The morning-evening difference of DBP was associated with AHI (r = -0.303, P = 0.011), amplitude of oxygen desaturation with pulse oxygen saturation (SpO(2)) less than 90% (OLA90%, r = -0.281, P = 0.018), and gradient of oxygen desaturation with SpO(2) less than 90% (OLG90%, r = 0.286, P = 0.035). Multiple linear regression analysis showed that, if AHI increased by 1/h, the morning-evening difference of DBP would increase 0.41 mm Hg; if the morning-evening difference of NO level in EBC increased by 1 micromol/L, the morning-evening difference of DBP would decrease 0.27 mm Hg; and if AHI increased by 1/h, the morning-evening difference of NO level in EBC would increase 0.40 micromol/L. CONCLUSION: The DBP of OSAHS patients in the morning is significantly higher than that in the evening. The morning-evening difference of NO level in EBC reflects indirectly the consumption of NO in the body, and is associated with the degree of OSAHS and the morning-evening difference of DBP.

[Investigation of Lung Cancer Patients Complicated with Chronic Obstructive Pulmonary Disease in Thoracic Surgical Department].

BACKGROUND: Lung cancer is an important complication of chronic obstructive pulmonary disease (COPD), and even significantly affects the prognosis of patients with COPD. COPD also affects the postoperative complications and recurrence in patients with lung cancer. This study aims to investigate lung cancer patients complicated with COPD in thoracic surgical department. METHODS: All medical records of lung cancer patients discharged from the Department of Thoracic Surgery of People's Hospital, Peking University during January 2015 and December 2015 were reviewed, including gender, age, tobacco smoke history, harmful occupational exposure, clinic symptom, chest computed tomography (CT) scanning, postoperative pathology result report, discharged diagnosis and spirometry [All patients underwent pulmonary function test are received bronchial dilation test if the based predicted value of forced expiratory volume in one second (FEV1) <70%]. RESULTS: A full set of lung function test was measured in 703 lung cancer patients. Bronchial dilation test was finished in 67 patients. 62 (92.5%) patients were diagnosed as COPD. 677 cases with lung cancer were received surgery. Bronchial dilation test was measured in 42 cases. Of them 38 (92.7%) patients were diagnosed as COPD. It was found that the patients with lung cancer and COPD was more frequent in males, elders (≥65 yr), smokers, non-adenocarcinoma patients than those of patients without COPD (P<0.05). The males and the elders (≥65 yr) were more likely to suffer from COPD (OR: 2.374-2.807, 95%CI: 1.101-7.157)(P<0.05). Only 3 patients (4.3‰) were diagnosed as COPD and received standard treatment before admission. And only 5 patients (7.1‰) were diagnosed as COPD as discharged. CONCLUSIONS: The routine pulmonary function as well as bronchial dilation test are helpful for screening the patients with COPD. At present, the diagnosis and treatment of lung cancer combined with COPD is a serious problem, which needs to be paid attention to by thoracic surgeons and to join hands with physicians in order to improve the diagnosis level of COPD.

[Crude extraction and activity assay of CEL I].

CEL I, extracted from celery, is the first known eukaryotic nuclease that cleaves DNA with high specificity at sites of base-substitution mismatch and DNA distortion. It is a key enzyme for TILLING research. Here we reported a crude extraction method and activity assay of CEL I. Incision at mismatches of single nucleotide suggested that CEL I can effectively detect DNA at G-->A base substitution and the result can be obtained from an ABI377 Sequencer. Therefore, the extracted enzyme can be used in TILLING.

[Genetic analysis and RAPD marker of the genes for brachytic stem trait in soybean].

Three crosses between NG94-156 (brachytic stem) and three varieties (normal stem) were made, and F2 segregative population and two recombined inbred line populations(F(7:8)) were obtained. Genetic analysis indicated that the brachytic stem of NG94-156 was controlled by two duplicate recessive genes. In searching for RAPD marker linked to the genes controlling brachytic stem, 260 RAPD primers were applied to screen four parents of three combinations and RIL. Polymorphic bands revealed by the primer S-506 exhibited the best repeatability among all primers. Linkage analysis indicated the genetic distance between S-506(1600) and brachytic stem gene was 6.94 cM.

Expression profile of hydrogen sulfide and its synthases correlates with tumor stage and grade in urothelial cell carcinoma of bladder.

BACKGROUND: Hydrogen sulfide (H2S) is a newly discovered gas transmitter. It is synthesized by cystathionine ß-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (MPST). Endogenous hydrogen sulfide has never been studied in bladder cancer. PURPOSE: We evaluated H2S production and its synthases expression levels in transitional cell carcinoma (urothelial cell carcinoma of bladder [UCB]) of human bladder tissue and cell lines. MATERIALS AND METHODS: Immunostaining was performed in urothelial cell lines and bladder specimens from 94 patients with UCB of different stages/grades. The expression levels/activities of CBS, CSE, and MPST of specimens and cell lines were analyzed by image semiquantity assay, western blot, and a sulfur-sensitive electrode. We tried to find the correlation between hydrogen sulfide and its synthases with tumor stage in UCB. All experiments were repeated at least 3 times. RESULTS: Immunoreactivity for CBS, CSE, and MPST was detected in malignant uroepithelium and muscular layer of all tissues examined and cultured cells. The expression levels of CBS, CSE, and MPST were associated with UCB stage/grade. Muscle-invasive bladder cancer samples showed the highest production of H2S (52.6±2.91 nmol/[mg·min]) among all tested samples and EJ cells (transitional cell carcinoma, grade IIIshowed the highest production of H2S among all tested cell lines (53.3±7.02nmol/[mg·min]). CONCLUSIONS: Protein levels and catalytic activities of CBS, CSE, and MPST increased with the increase of malignant degrees in human bladder tissues and human UCB cell lines. Our findings may promote the application of these novel enzymes to UCB diagnosis or treatment.

[Identification of drought tolerant germplasm and inheritance and QTL mapping of related root traits in soybean (Glycine max (L.) Merr.)].

Fifty nine accessions of soybean (Glycine max (L.) Merr.) selected from 301 ones in Huang-Huai-Hai and Middle-Lower Changjiang Valleys were tested in two years for their tolerance to drought by using the mean membership index value averaged over those of plant height,leave number,dry root weight and dry stem and leaf weight. Four most tolerant accessions (Rank 1) and two most sensitive ones (Rank 5) were identified. There existed very significant correlations between drought tolerance and relative values of dry root weight,total root length, and root volume (per plant dry weight basis), respectively,which could be used as root indicators of drought tolerance. The RIL population derived from Kefeng 1 x Nannong 1138-2 was used to analyze the inheritance of the three related root traits by using the segregation analysis of quantitative traits under the major gene plus polygene mixed inheritance model. The results showed that between the two parents (Rank 1 x Rank 4), the relative values of dry root weight,total root length and root volume were respectively controlled by two major genes (linked together for the latter two traits, recombination value being 4.30% and 1.93%, respectively) plus polygenes with their major gene heritability values of 62.26%-91.81% and polygene heritability values 2.99%-24.75%, indicating that the major genes,especially the one with larger effect,accounted for a major part of the genetic variation between the two parents. It was identified that five, three, and five QTLs located on N6-C2, N8-D1b + W, N11-E, and N18-K linkage groups for relative dry root weight, total root length and root volume, respectively. Each of the traits appeared to have one locus (Dw1, R/1, and Rv1) with relatively large effect in comparison with their other loci, and those major ones were located near the same site of the same linkage group N6-C2. The results of segregation analysis and QTL mapping appeared pretty consistent with each other, which could be used as a demonstration of each other.

[Segregation analysis of genetic system of quantitative traits in plants].

Based on the traditional polygene inheritance model of quantitative traits, the mixed major gene and polygene inheritance model was raised and considered as the general model, while pure major gene or pure polygene inheritance model being only the specific case of the general model. From the proposed theory, the segregation analysis procedure was established for studying the genetic system of quantitative traits of plants. At present this procedure can be used to evaluate the genetic effects of individual major genes (up to 2-3 major genes) and the collective genetic effects of polygenes as well as their heritability values. The present paper introduces the process of the establishment of the procedure, main achievements and application results. An example was given to illustrate the steps, methods and effectiveness of the procedure.

Inhibition of allergic responsiveness in a murine asthma model via IFN-gamma transgene expression.

OBJECTIVE: To investigate adenoviral vector mediated exogenous gene expression in mouse lungs and the effect of mIFN-gamma transgene expression on allergen-induced pulmonary eosinophil infiltration in a murine asthmatic model. METHODS: LacZ marker gene was transduced into CD-1 mouse airway epithelial cells by installation of a replication-deficient adenovirus with LacZ gene (AdCMVLacZ) 5 x 10(9) plaque forming unit (pfu) in the intratrachea or nostril. C57 mice were sensitized intraperitoneally and challenged by aerosol with ovalbumin (OVA) to produce an asthmatic model. AdCMVmIFNgamma 5 x 10(9) pfu was administered via nostril in asthmatic mice 48 h before OVA challenge. Sera, bronchial alveolar lavage (BAL) and lungs were recovered 48 h after OVA challenge. RESULTS: After administration with AdCMVLacZ by intratracheal installation or nose-drop, the lungs revealed a high level of widespread LacZ transduction with X-gal staining, mainly along airways. IFN-gamma via adenoviral vector transduction could be overexpressed both in vitro and in vivo (1624.7 +/- 1321.5 pg/ml in BAL 96 h after AdCMVIFNgamma infection). In AdCMVIFNgamma treated asthmatic models, histological evaluation revealed marked suppression of eosinophil peribronchial and perivascular infiltration; the recoverable percentage of eosinophils in BAL was an average of 9.00% +/- 4.58%, which was a statistically significant decrease versus that of the positive control group (75.13% +/- 6.85%) (P < 0.001). The total cell number in BAL ((145 +/- 55.6) x 10(3) cells/ml) in AdCMVmIFNgamma treated mice also was tremendously reduced compared to the positive control group ((216.6 +/- 71.1) x 10(3) cells/ml). CONCLUSIONS: Adenoviral vector was able to overexpress exogenous gene in murine lungs. IFN-gamma overexpression via adenoviral vector in pulmonary epithelia in vivo can abrogate allergen-induced eosinophilic infiltration in lungs in an asthmatic model, which may suggest a new preventively therapeutic method for cytokine immunogenetic transfer in allergic asthma.