[Clinical effect of negative pressure wound therapy in emergency limb-salvage operation of destructive injury of limb].

Objective: To explore the clinical effect of negative pressure wound therapy (NPWT) in emergency limb-salvage operation of destructive injury of limb. Methods: From July 2014 to December 2017, 43 patients with destructive injury of limb in one side conformed to the inclusion criteria were admitted to our hospital. The patients were divided to NPWT group of 24 patients [ 21 males and 3 females, aged (38±10) years] and routine dressing change group of 19 patients [ 17 males and 2 females, aged (37±10) years] according to their treatment methods. After the emergency debridement, fracture external fixation, neurovascular exploration, and microsurgical repair were performed, NPWT were applied on wounds of patients in NPWT group and routine dressing change treatment on wounds of patients in routine dressing change group. On 7 to 10 days after the emergency operation, incidence of arterial embolism of patients in the two groups were calculated, and condition of wound infection of patients in the two groups were observed. Complete wound healing time and survival condition of limb were recorded. Data were processed with independent sample t test or chi-square test. Results: Incidence of arterial embolism of patients in NPWT group on 7 to 10 days after the emergency operation was 6.67% (3/45), which was close to 5.56% (2/36) of patients in routine dressing change group (χ(2)=0.043, P>0.05). There was 1 patient with wound infection in NPWT group on 7 to 10 days after the emergency operation, obviously less than 6 patients in routine dressing change group (χ(2)=5.847, P<0.05). Complete wound healing time of patients in NPWT group was (30±4) d, significantly shorter than (36±8) d of patients in routine dressing change group (t=2.813, P<0.01). Limbs of 24 patients in NPWT group survived, which was close to 18 patients in routine dressing change group (χ(2)=1.293, P>0.05). Conclusions: NPWT can significantly reduce tthe wound infection rate and shorten the time of wound healing of limb with destructive injury after emergency operation, which is worthy of popularization in clinic.

[Application of adaptive statistical iterative reconstruction veo and 80 kv in renal computed tomography angiography].

Objective: To explore the application of adaptive statistical iterative reconstruction Veo (ASIR-V) and 80 kV in renal computed tomography angiography(CTA). Methods: Eighty patients with renal computed tomography angiography were prospectively collected from April 2018 to July 2018 in the Affiliated Hospital of Shaanxi University of Chinese Medicine and randomly divided into group A and group B. The patients in group A adopted tube voltage 120 kV and contrast agent concentration 600 mgI/kg and reconstructed with filtered back projection (FBP), while the patients in group B were scanned with tube voltage 80 kV and contrast agent concentration 350 mgI/kg and reconstructed with FBP and ASIR-V from 10% to 100% with 10% interval. The CT values and standard deviation (SD) of the right renal artery, left renal artery were measured respectively to calculate the signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR).The image quality of renal CTA was subjectively scored by two experienced radiologists blindly using a 5-point criteria.The contrast agent,CT volume dose index(CTDI(vol)) and dose length product(DLP) in both groups were recorded and the effective radiation dose(ED) was calculated. Results: The ED ((2.11±0.19)mSv) and contrast agent ((21.9±3.0)g) in group B were 65.1% (P<0.05) and 42.2% (P<0.05) lower than those in group A ((6.04±1.89)mSv and (38.0±3.8)g).With the increase of ASIR-V percentage in group B, CT values showed no significant difference, SD values gradually decreased, SNR values and CNR values gradually increased.The CT values with different reconstruction algorithm showed no statistically significant difference (all P>0.05) between group A and group B. The SD values with 40%ASIR-V to 100%ASIR-V reconstruction in group B were significantly lower than those of group A (all P<0.05).The SNR values with 50% ASIR-V to 100% ASIR-V reconstruction and CNR values with 70%ASIR-V to 100%ASIR-V were significantly higher than those of group A(all P<0.5).Two radiologists had excellent consistency in subjective scores of image quality for renal CTA(all kappa>0.75, P<0.05). The subjective scores with 60% ASIR-V to 90% ASIR-V in group B were significantly higher than those in group A (P<0.05), of which 70%ASIR-V reconstruction achieved the highest subjective score for renal CTA. Conclusion: ASIR-V and 80 kV can significantly reduce radiation dose (about 65.1%) and contrast agent (about 42.2%) in renal CTA, ASIR-V reconstruction can significantly improve the image quality of renal CTA, of which 70% ASIR-V reconstruction achieved the best image quality in 80 kV renal CTA.

[Surgical treatment of intractable odontoid fracture with atlantoaxial dislocation in children with spinal cord injury].

Objective: To discuss the surgical treatment and its effectiveness of odontoid fracture complicated with atlantoaxial dislocation in children with spinal cord injury. Methods: From January 2010 to December 2014, 10 cases of children under 14 years old, with intractable odontoid fracture with atlantoaxial dislocation were enrolled. The mean duration between injury and admission was 8.5 d (range 1-30 d). The surgery was performed using posterior reduction and internal fixation or anterior release combined with posterior fixation. The medical date including preoperative and postoperative neurological function, degree of reduction and fusion. Results: All cases were followed-up for 12-48 months. The last follow-up X ray and CT examinations showed good reduction and fusion. No fixation failure was observed among all the patients. According to the Frankel, the preoperative neurological function was C in 2 cases, D in 4 cases, and E in 4 cases. At the last follow-up, out of 2 cases with Frankel C improved to D, and the other 8 cases were back to normal. Conclusions: Diagnosis of odontoid fracture complicated with atlantoaxial dislocation is usually delayed in children. One-stage posterior reduction and internal fixation or anterior release combined with posterior fixation is an effective and safe surgical procedure for such cases.

Dynamic QTL analysis of protein content and glutamine synthetase activity in recombinant inbred wheat lines.

Protein content (PC) is a crucial factor that determines the end-use and nutritional quality of wheat (Triticum aestivum). Glutamine synthetase (GS), which is a major participant in nitrogen metabolism, can convert inorganic nitrogen into organic nitrogen. Although many studies have been conducted on PC and GS, a dynamic analysis of all of the filling stages has not been conducted. Therefore, 115 F9-10 recombinant inbred wheat lines of 'R131/R142' were used to analyze PC and GS activity during different developmental stages, using the conditional quantitative trait loci (QTL) mapping method. Twenty-two and six conditional QTL were detected for PC and GS activily, respectively. More QTL in leaf PC were detected during the early filling stages than in the later filling stages. Grain PC QTL displayed different dynamic variations to leaf PC QTL during the entire grain-filling stages. All of the QTL were expressed differently over time, and nine conditional QTL were detected across two filling stages. QTL with similar functions may have tended to group in specific locales. This study provides dynamic genetic information on protein accumulation during grain-filling stages.

Identification and genetic mapping of the putative Thinopyrum intermedium-derived dominant powdery mildew resistance gene PmL962 on wheat chromosome arm 2BS.

KEY MESSAGE: Powdery resistance putatively derived from Thinopyrum intermedium in the wheat line L962 is controlled by a single dominant gene designated PmL962 and mapped to chromosome arm 2BS. Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive disease affecting the production of wheat (Triticum aestivum). Powdery mildew resistance was putatively transferred from Thinopyrum intermedium to the common wheat line L962, which conferred resistance to multiple Chinese Bgt isolates. Genetic analysis of the powdery mildew response was conducted by crossing the resistant line L962 with the susceptible line L983. Disease assessments of the F1, F2, and F2:3 populations from the cross L983/L962 indicated that resistance was controlled by a single dominant gene. A total of 373 F2:3 lines and 781 pairs of genomic simple sequence repeat (SSR) primers were employed to determine the chromosomal location of the resistance gene. The gene was linked to four publicly available and recently developed wheat genomic SSR markers and seven EST-STS markers. The resistance gene was mapped to chromosome arm 2BS based on the locations of the linked markers. Pedigree, molecular marker and resistance response data indicated that the powdery mildew resistance gene in L962 is novel. It was temporarily designated PmL962. It is flanked by Xwmc314 and BE443737at genetic distances of 2.09 and 3.74 cM, respectively, and located in a 20.77 cM interval that is co-linear with a 269.4 kb genomic region on chromosome 5 in Brachypodium distachyon and a 223.5 kb genomic region on rice (Oryza sativa) chromosome 4. The markers that are closely linked to this gene have potential applications in marker-assisted breeding.

Diversity resistance to Puccinia striiformis f. sp Tritici in rye chromosome arm 1RS expressed in wheat.

The 1BL.1RS wheat-rye translocation contained in the Russian cultivar Aurora has been the most widespread alien translocation in wheat-breeding programs all over the world. However, following the prevalence of new biotypes of the pathogens, disease-resistance genes in this translocation chromosome have been overcome and consequently they have been eliminated in modern wheat-breeding programs. In this paper, we report on 12 new primary 1BL.1RS translocation lines derived from the crosses of a Chinese high yield wheat cv. Mianyang 11 with three rye cultivars collected from China. GISH, C-banding and PCR techniques using the specific primers for 1BS, 1RS and centromeres of wheat and rye were applied to identify the constitution of chromosomes. The results confirmed that all 1BL.1RS chromosomes in the 12 primary translocation lines contained integrated 1RS chromosome arms. In the resistance analysis using five kinds of Pst pathotypes, the 12 primary translocation lines showed diversity resistance to stripe rust, which contained at least five different new genes (alleles), significantly different from the Yr9 gene coming from Russian wheat cultivar Aurora. The results indicated that the chromosome arm 1RS in the rye population carries abundant yet untapped genes (alleles) for resistance to wheat stripe rust, which would originate from the neutral diversity in the natural population of rye. It is suggested that creating more primary translocation lines in genome modification will be extremely important to use the diversity of alien R-genes, which was generated by long-term neutral mutation and maintained in the population of alien species.

Characterization of a new wheat-Aegilops biuncialis addition line conferring quality-associated HMW glutenin subunits.

In this study, a new disomic addition line, 12-5-2, with 44 chromosomes that was derived from BC3F2 descendants of the hybridization between Triticum aestivum cv. CN19 and Aegilops biuncialis was created and reported. 12-5-2 was immune to both powdery mildew and stripe rust and has stable fertility. Fluorescence in situ hybridization and C-banding revealed that 12-5-2 was a 1U(b) disomic addition line (ADL1U(b)). The seed storage protein electrophoresis showed that 12-5-2 presented all high molecular weight glutenin subunits (7 + 8 and 2 + 12) of CN19 and 2 new subunits that were designated Ux and Uy. Additionally, the flour quality parameters showed that the protein content, Zeleny sedimentation value, wet gluten content, and grain hardness of 12-5-2 were significantly higher than those of its parent CN19. Moreover, 5 pairs of the chromosome 1U(b)-specific polymerase chain reaction-based landmark unique gene markers, TNAC1021, TNAC1041, TNAC1071, TNAC1-01, and TNAC1-04, were also obtained. The new ADL1U(b) 12-5-2 could be a valuable source for wheat improvement, especially for wheat end-product quality and resistance to disease.

Genetic mapping of a putative Thinopyrum intermedium-derived stripe rust resistance gene on wheat chromosome 1B.

KEY MESSAGE: Stripe rust resistance transferred from Thinopyrum intermedium into common wheat was controlled by a single dominant gene, which mapped to chromosome 1B near Yr26 and was designated YrL693. Stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) is a highly destructive disease of wheat (Triticum aestivum). Stripe rust resistance was transferred from Thinopyrum intermedium to common wheat, and the resulting introgression line (L693) exhibited all-stage resistance to the widely virulent and predominant Chinese pathotypes CYR32 and CYR33 and to the new virulent pathotype V26. There was no cytological evidence that L693 had alien chromosomal segments from Th. intermedium. Genetic analysis of stripe rust resistance was performed by crossing L693 with the susceptible line L661. F(1), F(2), and F(2:3) populations from reciprocal crosses showed that resistance was controlled by a single dominant gene. A total 479 F(2:3) lines and 781 pairs of genomic simple sequence repeat (SSR) primers were employed to determine the chromosomal location of the resistance gene. The gene was linked to six publicly available and three recently developed wheat genomic SSR markers. The linked markers were localized to wheat chromosome 1B using Chinese Spring nulli-tetrasomic lines, and the resistance gene was localized to chromosome 1B based on SSR and wheat genomic information. A high-density genetic map was also produced. The pedigree, molecular marker data, and resistance response indicated that the stripe rust resistance gene in L693 is a novel gene, which was temporarily designated YrL693. The SSR markers that co-segregate with this gene (Xbarc187-1B, Xbarc187-1B-1, Xgwm18-1B, and Xgwm11-1B) have potential application in marker-assisted breeding of wheat, and YrL693 will be useful for broadening the genetic basis of stripe rust resistance in wheat.

Major quality trait analysis and QTL detection in hexaploid wheat in humid rain-fed agriculture.

Humid rain-fed agriculture is a special environment for wheat (Triticum aestivum) culture that tends to negatively affect wheat yield and quality. To identify quality characters of wheat in a humid environment, we conducted quality analysis and quantitative trait loci (QTL) detection in a recombinant inbred line whose parent had a high level of quality for several years. We found that high-quality wheat had less gluten content and lower protein content. Apparently, wheat quality and associated quantity traits were in a dynamic state of equilibrium. We detected 83 QTL for 10 wheat quality traits in this recombinant inbred line population. Nine QTL were detected in both evaluation years; Q.DT.scau-2A, linked to Xwmc522-2A, was detected at the same genetic location in both years. Other QTL for different traits were detected simultaneously in more than one location. Consequently, there appeared to be pleiotropic genes that control wheat quality. Based on previous studies and our research on QTL analysis of grain protein content, we conclude that there must be one or more genes for grain protein content on chromosome 6B, whose expression was little affected by environment. We constructed a consensus map and projected the QTL on it. It was useful for choosing optimal markers for marker-assisted breeding and map-based cloning.

QTL analysis of the spring wheat "Chapio" identifies stable stripe rust resistance despite inter-continental genotype × environment interactions.

Chapio is a spring wheat developed by CIMMYT in Mexico by a breeding program that focused on multigenic resistances to leaf rust and stripe rust. A population consisting of 277 recombinant inbred lines (RILs) was developed by crossing Chapio with Avocet. The RILs were genotyped with DArT markers (137 randomly selected RILs) and bulked segregant analysis conducted to supplement the map with informative SSR markers. The final map consisted of 264 markers. Phenotyping against stripe rust was conducted for three seasons in Toluca, Mexico and at three sites over two seasons (total of four environments) in Sichuan Province, China. Significant loci across the two inter-continental regions included Lr34/Yr18 on 7DS, Sr2/Yr30 on 3BS, and a QTL on 3D. There were significant genotype × environment interactions with resistance gene Yr31 on 2BS being effective in most of the Toluca environments; however, a late incursion of a virulent pathotype in 2009 rendered this gene ineffective. This locus also had no effect in China. Conversely, a 5BL locus was only effective in the Chinese environments. There were also complex additive interactions. In the Mexican environments, Yr31 suppressed the additive effect of Yr30 and the 3D locus, but not of Lr34/Yr18, while in China, the 3D and 5BL loci were generally not additive with each other, but were additive when combined with other loci. These results indicate the importance of maintaining diverse, multi-genic resistances as Chapio had stable inter-continental resistance despite the fact that there were QTLs that were not effective in either one or the other region.

Chloroplast ultrastructure regeneration with protection of photosystem II is responsible for the functional 'stay-green' trait in wheat.

CN17 is a functional stay-green wheat variety that exhibits delayed leaf senescence and enhanced photosynthetic competence. To better understand these valuable traits, levels of chlorophyll a and b, soluble proteins, unsaturated fatty acids, and other components of CN17 were assayed. In addition, chloroplast ultrastructure, chloroplast number, and differences in gene expression between CN17 and a control variety, MY11, were examined. By 21 d post-anthesis (DPA), CN17 leaves exhibited a significantly higher maximal photochemical efficiency for photosystem II (PSII) (F(v) /F(m) ) and a significantly higher efficiency of excitation capture by open PSII reaction centres (F(v) '/F(m) '). In addition, chlorophyll degradation in CN17 was delayed by approximately 14 d, and was not blocked as observed in cosmetic stay-green phenotypes. The soluble protein content (Ps) of CN17 was higher than MY11 at all timepoints assayed, and the ratio of unsaturated to saturated fatty acids was significantly higher. CN17 also exhibited isolated granal lamellae associated with vesicles and diminished peroxidation, and between 35 and 42 DPA, a sharp decrease in chloroplast number was detected. Taken together, these results strongly support the hypothesis that chloroplast ultrastructure regeneration is responsible for the functional stay-green trait of CN17, and gene expression data provide insight into the mechanistic details.

Molecular cytogenetic characterization of a new leaf rolling triticale.

Leaf rolling occurs in some cereal genotypes in response to drought. We identified and made a phenotypic, cytological and physiological analysis of a leaf-rolling genotype (CMH83) of hexaploid triticale (X Triticosecale Wittmack) that exhibited reduced plant height, rolled and narrow leaves. Gliadin electrophoresis of seed protein showed that CMH83 was genetically stable. Sequential Giemsa-C-banding and genomic in situ hybridization showed that CMH83 contains 12 rye chromosomes; two pairs of these chromosomes have reduced telomeric heterochromatin bands. Tests of relative water content and water loss rate of leaves of CMH83 compared with those of wheat cultivars indicated that rapid water loss after drought stress in CMH83 is associated with the leaf rolling phenotypes. Leaf rolling in CMH83 was a dominant trait in our inheritance studies. Triticale line CMH83 could be used to study drought resistance mechanisms in triticale.

Evolutionary trends of microsatellites during the speciation process and phylogenetic relationships within the genus Secale.

Eleven weedy or wild species or subspecies of the genus Secale L. were compared with a set of cultivated rye accessions, based on inter-simple sequence repeat (ISSR) markers to analyze their phylogenetic relationships. A total of 846 bands were amplified from reactions using 12 screening primers, including 79 loci with a mean of 10.1 alleles per locus. The number of amplified bands for each primer ranged from 12 to 134, with a mean of 70.5 amplified bands per primer. The presence and distribution of amplified bands in different accessions demonstrate that a rapid evolutionary trend of microsatellite repeats occurred during the speciation process from the perennial wild form to annual cultivated rye. In addition, variation, amplification, and deletion of microsatellites in genomes revealed phylogenetic relationships in the genus Secale. Analysis of the presence, number, and distribution of amplified bands in genomes, as well as the comparison with genetic similarity (GS) indices based on ISSR, indicate that Secale strictum subsp. africanum (Stapf) Hammer, Secale strictum anatolicum (Boiss.) Hammer, Secale sylvestre Host, and Secale strictum subsp. strictum (C. Presl) Hammer emerged in succession from a common ancestor of Secale following geographic separation and genetic differentiation. The annual weedy rye evolved from S. strictum subsp. strictum, which was domesticated as present-day cultivated rye. Data from ISSR analyses separated all investigated accessions of the genus Secale into three distinct groups. These results support the division of the genus Secale into three species: the annual wild species S. sylvestre; the perennial wild species S. strictum, including several differential subspecies forms such as strictum, africanum, and anatolicum; and S. cereale, including cultivated and weedy rye as subspecies forms.

Characterization and chromosomal location of Pm40 in common wheat: a new gene for resistance to powdery mildew derived from Elytrigia intermedium.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a very destructive wheat (Triticum aestivum) disease. Resistance was transferred from Elytrigia intermedium to common wheat by crossing and backcrossing, and line GRY19, that was subsequently selected, possessed a single dominant gene for seedling resistance. Five polymorphic microsatellite markers, Xgwm297, Xwmc335, Xwmc364, Xwmc426 and Xwmc476, on chromosome arm 7BS, were mapped relative to the powdery mildew resistance locus in an F(2) population of Mianyang 11/GRY19. The loci order Xwmc426-Xwmc335-Pm40-Xgwm297-Xwmc364-Xwmc476, with 5.9, 0.2, 0.7, 1.2 and 2.9 cM genetic distances, was consistent with published maps. The resistance gene transferred from Elytrigia intermedium into wheat line GRY19 was novel, and was designated Pm40. The close flanking markers will enable marker assisted transfer of this gene into wheat breeding populations.

Allelic analysis of stripe rust resistance genes on wheat chromosome 2BS.

Stripe rust, caused by Puccinia striiormis Westend f. sp. tritici, is one of the most important foliar diseases of wheat (Triticum aestivum L.) worldwide. Stripe rust resistance genes Yr27, Yr31, YrSp, YrV23, and YrCN19 on chromosome 2BS confer resistance to some or all Chinese P. striiormis f. sp. tritici races CYR31, CYR32, SY11-4, and SY11-14 in the greenhouse. To screen microsatellite (SSR) markers linked with YrCN19, F1, F2, and F3 populations derived from cross Ch377/CN19 were screened with race CYR32 and 35 SSR primer pairs. Linkage analysis indicated that the single dominant gene YrCN19 in cultivar CN19 was linked with SSR markers Xgwm410, Xgwm374, Xwmc477, and Xgwm382 on chromosome 2BS with genetic distances of 0.3, 7.9, 12.3, and 21.2 cM, respectively. Crosses of CN19 with wheat lines carrying other genes on chromosome 2B showed that all were located at different loci. YrCN19 is thus different from the other reported Yr genes in chromosomal location and resistance response and was therefore named Yr41. Prospects and strategies of using Yr41 and other Yr genes in wheat improvement for stripe rust resistance are discussed.

Identification, Chromosome Location, and Diagnostic Markers for a New Gene (YrCN19) for Resistance to Wheat Stripe Rust.

ABSTRACT Several wheat lines and cultivars of wheat (Triticum aestivum) originating from the southwestern region of China were found to be highly resistant to stripe rust by inoculation with the prevalent races (CYR30, CYR31, and CYR32) and newly emerged races (H46-4, SY11-4 and SY11-14) of the pathogen. An inheritance study of the resistance to stripe rust was carried out by crossing resistant AIM6 with susceptible BeiZ76. Results indicated that the resistance to stripe rust was controlled by a single dominant gene. The 112 F(2) plants chosen from the cross BeiZ76/ AIM6 were analyzed with 218 pairs of microsatellite primers to determine the map location of the resistance gene. A simple sequence repeat marker on chromosome arm 2BS, Xgwm410, showed polymorphism and co-segregation between stripe rust resistant and susceptible plants. From the pedigree, inheritance, molecular marker, and resistance response, it is concluded that the stripe rust resistance gene in wheat cv. Chuan-nong19 (CN19) and wheat lines AIM5 and AIM6 is a novel gene, designated YrCN19. The microsatellite primer Xgwm410 is a diagnostic marker of the resistance gene YrCN19, which has potential for application in the marker-assisted breeding of wheat.

Hybrid necrosis in triticale and the expression of necrosis genes in allopolyploids.

The occurrence in triticale of four different genes causing hybrid necrosis is described: Ne1 and Ne2 in the B genome of wheat and Ner1 and Ner2 in the rye genome. Hybrid necrosis develops due to dominant complementary interaction of two genes. This interaction in triticale, however, may take place not only between genes belonging to the same genome but also between genes of different genomes. In triticale, these genes can cause hybrid necrosis in four different combinations. The inheritance of the phenomenon in triticale is, therefore, more complicated than it is in wheat or rye. To avoid hybrid necrosis in triticale, attention should be paid that no necrosis genes are introduced into the primary triticale stocks from the wheat and rye parents. The expression of necrosis genes is influenced by the level of ploidy. Any additional genome - A, B, D, or R - may exert a suppressing effect on the expression of necrosis genes. Therefore, when identifying genotypes of triticale with regard to their necrosis genes, the level of ploidy has to be accounted for. Moreover, the present results illustrate that gene expression in polyploids is not only determined by interactions with other single genes but that it may also be modified by the total genotype of the respective individual.