[Establishment and validation of a novel nomogram to predict overall survival after radical nephrectomy].

Objective: To establish a nomogram prognostic model for predicting the 5-, 10-, and 15-year overall survival (OS) of non-metastatic renal cell carcinoma patients managed with radical nephrectomy (RN), compare the modelled results with the results of pure pathologic staging, the Karakiewicz nomogram and the Mayo Clinic Stage, Size, Grade, and Necrosis (SSIGN) score commonly used in foreign countries, and stratify the patients into different prognostic risk subgroups. Methods: A total of 1 246 non-metastatic renal cell carcinoma patients managed with RN in Sun Yat-sen University Cancer Center (SYSUCC) from 1999 to 2020 were retrospectively analyzed. Multivariate Cox regression analysis was used to screen the variables that influence the prognosis for nomogram establishment, and the bootstrap random sampling was used for internal validation. The time-receiver operating characteristic curve (ROC), the calibration curve and the clinical decision curve analysis (DCA) were applied to evaluate the nomogram. The prediction efficacy of the nomogram and that of the pure pathologic staging, the Karakiewicz nomogram and the SSIGN score was compared through the area under the curve (AUC). Finally, patients were stratified into different risk subgroups according to our nomogram scores. Results: A total of 1 246 patients managed with RN were enrolled in this study. Multivariate Cox regression analysis showed that age, smoking history, pathological nuclear grade, sarcomatoid differentiation, tumor necrosis and pathological T and N stages were independent prognostic factors for RN patients (all P<0.05). A nomogram model named SYSUCC based on these factors was built to predict the 5-, 10-, and 15-year survival rate of the participating patients. In the bootstrap random sampling with 1 000 iterations, all these factors occurred for more than 800 times as independent predictors. The Harrell's concordance index (C-index) of SYSUCC was higher compared with pure pathological staging [0.770 (95% CI: 0.716-0.823) vs 0.674 (95% CI: 0.621-0.728)]. The calibration curve showed that the survival rate as predicted by the SYSUCC model simulated the actual rate, while the clinical DCA showed that the SYSUCC nomogram has a benefit in certain probability ranges. In the ROC analysis that included 857 patients with detailed pathological nuclear stages, the nomogram had a larger AUC (5-/10-year AUC: 0.823/0.804) and better discriminating ability than pure pathological staging (5-/10-year AUC: 0.701/0.658), Karakiewicz nomogram (5-/10-year AUC: 0.772/0.734) and SSIGN score (5-/10-year AUC: 0.792/0.750) in predicting the 5-/10-year OS of RN patients (all P<0.05). In addition, the AUC of the SYSUCC nomogram for predicting the 15-year OS (0.820) was larger than that of the SSIGN score (0.709), and there was no statistical difference (P<0.05) between the SYSUCC nomogram, pure pathological staging (0.773) and the Karakiewicz nomogram (0.826). The calibration curve was close to the standard curve, which indicated that the model has good predictive performance. Finally, patients were stratified into low-, intermediate-, and high-risk subgroups (738, 379 and 129, respectively) according to the SYSUCC nomogram scores, among whom patients in intermediate- and high-risk subgroups had a worse OS than patients in the low-risk subgroup (intermediate-risk group vs. low-risk group: HR=4.33, 95% CI: 3.22-5.81, P<0.001; high-risk group vs low-risk group: HR=11.95, 95% CI: 8.29-17.24, P<0.001), and the high-risk subgroup had a worse OS than the intermediate-risk group (HR=2.63, 95% CI: 1.88-3.68, P<0.001). Conclusions: Age, smoking history, pathological nuclear grade, sarcomatoid differentiation, tumor necrosis and pathological stage were independent prognostic factors for non-metastasis renal cell carcinoma patients after RN. The SYSUCC nomogram based on these independent prognostic factors can better predict the 5-, 10-, and 15-year OS than pure pathological staging, the Karakiewicz nomogram and the SSIGN score of patients after RN. In addition, the SYSUCC nomogram has good discrimination, agreement, risk stratification and clinical application potential.

[Long-term survival analysis of 1 367 patients treated with radical nephrectomy from a single center].

Objective: To report the long-term survival of renal cell carcinoma (RCC) patients treated with radical nephrectomy in Sun Yat-sen University Cancer Center. Methods: We retrospectively analyzed the clinical, pathological and follow-up records of 1 367 non-metastatic RCC patients treated with radical nephrectomy from 1999 to 2020 in this center. The primary endpoint of this study was overall survival rate. Survival curves were estimated using the Kaplan-Meier method, and group differences were compared through Log-rank test. Univariate and multivariate Cox analysis were fit to determine the clinical and pathological features associated with overall survival rate. Results: A total of 1 367 patients treated with radical nephrectomy with complete follow-up data were included in the study. The median follow-up time was 52.6 months, and 1 100 patients survived and 267 died, with the median time to overall survival not yet reached. The 5-year and 10-year overall survival rates were 82.8% and 74.9%, respectively. The 5-year and 10-year overall survival rates of Leibovich low-risk patients were 93.3% and 88.2%, respectively; of Leibovich intermediate-risk patients were 82.2% and 72.3%, respectively; and of Leibovich high-risk patients were 50.5% and 30.2%, respectively. There were significant differences in the long-term survival among the three groups (P<0.001). The 10-year overall survival rates for patients with pT1, pT2, pT3 and pT4 RCC were 83.2%, 73.6%, 55.0% and 31.4%, respectively. There were significant differences among pT1, pT2, pT3 and pT4 patients(P<0.001). The 5-year and 10-year overall survival rates of patients with lymph node metastasis were 48.5% and 35.6%, respectively, and those of patients without lymph node metastasis were 85.1% and 77.5%, respectively. There was significant difference in the long-term survival between patients with lymph node metastasis and without lymph node metastasis. The 10-year overall survival rate was 96.2% for nuclear Grade 1, 81.6% for nuclear Grade 2, 60.5% for nuclear Grade 3, and 43.4% for nuclear Grade 4 patients. The difference was statistically significant. There was no significant difference in the long-term survival between patients with localized renal cancer (pT1-2N0M0) who underwent open surgery and minimally invasive surgery (10-year overall survival rate 80.5% vs 85.6%, P=0.160). Multivariate Cox analysis showed that age≥55 years (HR=2.11, 95% CI: 1.50-2.96, P<0.001), T stage(T3+ T4 vs T1a: HR=2.37, 95% CI: 1.26-4.46, P=0.008), local lymph node metastasis (HR=3.04, 95%CI: 1.81-5.09, P<0.001), nuclear grade (G3-G4 vs G1: HR=4.21, 95%CI: 1.51-11.75, P=0.006), tumor necrosis (HR=1.66, 95% CI: 1.17-2.37, P=0.005), sarcomatoid differentiation (HR=2.39, 95% CI: 1.31-4.35, P=0.005) and BMI≥24kg/m(2) (HR=0.56, 95%CI: 0.39-0.80, P=0.001) were independent factors affecting long-term survival after radical nephrectomy. Conclusions: The long-term survival of radical nephrectomy in patients with renal cell carcinoma is satisfactory. Advanced age, higher pathological stage and grade, tumor necrosis and sarcomatoid differentiation were the main adverse factors affecting the prognosis of patients. Higher body mass index was a protective factor for the prognosis of patients.

[Relation factor analysis for the short-term preservation of ipsilateral renal function after partial nephrectomy].

Objectives: To analyze the factors relative to the short-term preservation of ipsilateral renal function after partial nephrectomy. Methods: The clinical data of 83 patients who were treated with partial nephrectomy from December 2014 to December 2019 in the Department of Urology, Sun Yat-sen University Cancer Center were retrospectively analyzed. There were 54 males and 29 females, aging (M (IQR)) 49 (17) years (range: 27 to 74 years). The ischemia time in operation was 25 (18) minutes (range: 10 to 67 minutes). Emission computed tomography scan and CT scan were performed before (within 1 month) and after (3 to 12 months) surgery. The volume of the ipsilateral and contralateral kidney was measured on the basis of preoperative and postoperative CT scans. The glomerular filtration rate (GFR) specifically in each kidney was estimated by emission computed tomography. Recovery from ischemia is determined by the formula: GFR preservation/volume saved×100%. Linear regression was used to explore the factors ralative to the short-term preservation of ipsilateral renal function after partial nephrectomy. Results: The GFR preservation of the ipsilateral kidney was 80.9 (25.2) % (range: 31.0% to 109.4%). The volume loss of the kidney resulted in a decrease of 12.0% (5.8 ml/(min×1.96 m2)) of GFR, while the ischemic injury resulted in a decrease of 6.5% (2.5 ml/(min×1.96 m2)) of GFR. The volume saved from the ipsilateral kidney was 87.1 (12.9) % (range: 27.0% to 131.7%). Recovery from ischemia was 93.5 (17.5) % (range:44.3% to 178.3%). In multivariate analysis, GFR preservation of the ipsilateral kidney was significantly correlated with the volume saved of the ipsilateral kidney (ß=0.383, 95%CI: 0.144 to 0.622, P=0.002). It was not related to the ischemia time (ß=0.046, 95%CI:-0.383 to 0.475, P=0.831). Conclusion: In the condition of limited ischemic time, in the short term ipsilateral renal function after partial nephrectomy is mainly determined by the loss of kidney volume, while ischemic injury only plays a minor role.

[Efficacy of partial nephrectomy in patients with localized renal carcinoma: a 20-year experience of 2 046 patients in a single center].

Objectives: To analyze the long-term survival of patients with localized renal cell carcinoma after partical nephrectomy. Methods: The clinicopathological records and survival follow-up data of 2 046 patients with localized renal cell carcinoma, who were treated with partial nephrectomy from August 2001 to February 2021 in the Department of Urology, Sun Yat-sen University Cancer Center, were retrospectively analyzed. There were 1 402 males and 644 females, aged (M(IQR)) 51 (19) years (range: 6 to 86 years). The primary end point of this study was cancer-specific survival. Survival curves were estimated using the Kaplan-Meier method, and the difference test was performed by Log-rank test. Univariate and multivariate Cox analysis were fitted to determine factors associated with cancer-specific survival. Results: The follow-up time was 49.2 (48.0) months (range: 1 to 229 months), with 1 974 patients surviving and 72 dying. The median cancer-specific survival time has not yet been reached. The 5- and 10-year cancer specific survival rates were 97.0% and 91.2%, respectively. The 10-year cancer-specific survival rates for stage pT1a (n=1 447), pT1b (n=523) and pT2 (n=58) were 95.3%, 81.8%, and 81.7%, respectively. The 10-year cancer-specific survival rates of patients with nuclear grade 1 (n=226), 2 (n=1 244) and 3 to 4 (n=278) were 96.6%, 89.4%, and 85.5%, respectively. There were no significant differences in 5-year cancer-specific survival rates among patients underwent open, laparoscopic, or robotic surgery (96.7% vs. 97.1% vs. 97.5%, P=0.600). Multivariate analysis showed that age≥50 years (HR=3.93, 95%CI: 1.82 to 8.47, P<0.01), T stage (T1b vs. T1a: HR=3.31, 95%CI: 1.83 to 5.99, P<0.01; T2+T3 vs. T1a: HR=2.88, 95%CI: 1.00 to 8.28, P=0.049) and nuclear grade (G3 to 4 vs. G1: HR=2.81, 95%CI: 1.01 to 7.82, P=0.048) were independent prognostic factors of localized renal cell carcinoma after partial nephrectomy. Conclusions: The long-term cancer-specific survival rates of patients with localized renal cancer after partial nephrectomy are satisfactory. The type of operation (open, laparoscopic, or robotic) has no significant effect on survival. However, patients with older age, higher nuclear grade, and higher T stage have a lower cancer-specific survival rate. Grasping surgical indications, attaching importance to preoperative evaluation, perioperative management, and postoperative follow-up, could benefit achieving satisfactory long-term survival.

[Effects of three methods for correction of low-degree against-the-rule corneal astigmatism during cataract surgery].

Objective: To compare the clinical effects of and visual quality after correction of low-degree against-the-rule (ATR) corneal astigmatism by implantation of an astigmatism-corrected intraocular lens (IOL), femtosecond laser release and manual release in cataract surgery. Methods: It was a prospective cohort study. A total of 120 patients (120 eyes) with cataract combined with low-degree ATR corneal astigmatism diagnosed in Chongqing Aier Mega Eye Hospital from December 2017 to October 2020 were included and divided into 3 groups, each with 40 patients, according to their own selections of astigmatism correction methods during cataract surgery. In the astigmatism-corrected IOL group, phacoemulsification for cataract extraction combined with toric IOL implantation was performed. In the femtosecond laser release group, astigmatic keratotomy using a femtosecond laser was combined. In the manual release group, a limbal relaxing incision was made. Uncorrected distance visual acuity (UDVA) and corneal astigmatism were measured before surgery. At 3 months and 1 year after surgery, UDVA and best-corrected distance visual acuity were examined, as well as whole eye residual astigmatism by ARK-1, corneal astigmatism by the IOLMaster 500, whole eye high order aberration (HOA) and modulation transfer function (MTF) by the iTrace visual function analyzer. Analysis of variance was used for the comparison of data in a normal distribution. Repeated measures were used for the comparison within groups. The rank sum test was used for the comparison of data that were not normally distributed. Results: Of the 120 patients, 100 patients (100 eyes), including 44 males and 56 females, with an age of (66.48±6.20) years, completed the follow-up. Among the three groups, the differences were not statistically significant in terms of gender distribution, age, preoperative corneal astigmatism, UDVA and spherical equivalent of the IOL (all P>0.05). At 3 months and 1 year after surgery, the UDVA was significantly better than that before surgery in each group (Z=5.18, 5.04, 4.98, 4.99, 4.90, 4.89; all P<0.001). At the two time points, the differences in the whole eye residual astigmatism among the three groups were statistically significant (H=30.69, 31.23; both P<0.001). At 3 months, the whole eye residual astigmatism in the astigmatism-corrected IOL group was lower than that in the other two groups. At 1 year, the residual astigmatism in the astigmatism-corrected IOL group [0.25(0.00, 0.50) D] was also lower compared to that in the femtosecond laser release group [0.50(0.50, 0.75) D] and the manual release group [0.75(0.50, 0.75) D] (Z=-3.71, -5.18, -3.94, -5.15; all P<0.001). The differences in the HOA at 3 months and 1 year among the three groups were statistically significant (H=36.30, 34.38; both P<0.001). The HOA in the astigmatism-corrected IOL group was significantly higher than that in the other two groups at the two time points (Z=5.01, 4.73, 5.31, 5.27; all P<0.001). At 3 months and 1 year, the differences in the MTF value among the three groups were also statistically significant (H=30.02, 29.92; both P<0.001), and the MTF value in the femtosecond laser release group was significantly higher than that in the other two groups (Z=4.61, 4.67, 4.66, 4.69; all P<0.001). Conclusions: All the three astigmatism correction methods used at the time of cataract surgery can effectively correct low-degree ATR corneal astigmatism. The residual astigmatism in the whole eye after astigmatism-corrected IOL implantation is small and stable, while the HOA after release using the femtosecond laser is low with good visual quality.

Selection of Differential Isolates of Magnaporthe oryzae for Postulation of Blast Resistance Genes.

A set of differential isolates of Magnaporthe oryzae is needed for the postulation of blast resistance genes in numerous rice varieties and breeding materials. In this study, the pathotypes of 1,377 M. oryzae isolates from different regions of China were determined by inoculating detached rice leaves of 24 monogenic lines. Among them, 25 isolates were selected as differential isolates based on the following characteristics: they had distinct responses on the monogenic lines, contained the minimum number of avirulence genes, were stable in pathogenicity and conidiation during consecutive culture, were consistent colony growth rate, and, together, could differentiate combinations of the 24 major blast resistance genes. Seedlings of rice cultivars were inoculated with this differential set of isolates to postulate whether they contain 1 or more than 1 of the 24 blast resistance genes. The results were consistent with those from polymerase chain reaction analysis of target resistance genes. Establishment of a standard set of differential isolates will facilitate breeding for blast resistance and improved management of rice blast disease.

Kisspeptin-13 enhances memory and mitigates memory impairment induced by Aß1-42 in mice novel object and object location recognition tasks.

Kisspeptin (KP), the endogenous ligand of GPR54, is a recently discovered neuropeptide shown to be involved in regulating reproductive system, anxiety-related behavior, locomotion, food intake, and suppression of metastasis across a range of cancers. KP is transcribed within the hippocampus, and GPR54 has been found in the amygdala and hippocampus, suggesting that KP might be involved in mediating learning and memory. However, the role of KP in cognition was largely unclear. Here, we investigated the role of KP-13, one of the endogenous active isoforms, in memory processes, and determined whether KP-13 could mitigate memory impairment induced by Aß1-42 in mice, using novel object recognition (NOR) and object location recognition (OLR) tasks. Intracerebroventricular (i.c.v.) infusion of KP-13 (2µg) immediately after training not only facilitated memory formation, but also prolonged memory retention in both tasks. The memory-improving effects of KP-13 could be blocked by the GPR54 receptor antagonist, kisspeptin-234 (234), and GnRH receptors antagonist, Cetrorelix, suggesting pharmacological specificity. Then the memory-enhancing effects were also presented after infusion of KP-13 into the hippocampus. Moreover, we found that i.c.v. injection of KP-13 was able to reverse the memory impairment induced by Aß1-42, which was inhibited by 234. To sum up, the results of our work indicate that KP-13 could facilitate memory formation and prolong memory retention through activation of the GPR54 and GnRH receptors, and suppress memory-impairing effect of Aß1-42 through activation of the GPR54, suggesting that KP-13 may be a potential drug for enhancing memory and treating Alzheimer's disease.

Differential expression of CYB5A in Chinese and European pig breeds due to genetic variations in the promoter region.

Cytochrome b5 (CYB5A) is an important electron transfer protein with homologues in a number of different organisms. In pigs, CYB5A is related to boar taint because of its role in androstenone biosynthesis. To determine the variety of CYB5A expression in pig breeds, genetic variations in the porcine CYB5A promoter region in both Chinese and European pig breeds were examined. Three single nucleotide polymorphisms (NC_010443.4:g.165901487delG, g.165901767T>C and g.165902078C>T) were identified in the porcine CYB5A promoter region. These SNPs occurred in different frequencies in Chinese and European pigs. Chinese pigs were primarily haplotype B (denoted as delG-C-T: the position of nt 165901487 of the CYB5 gene is a G deletion, nt 165901767 is C and nt 165902078 is T), except for Licha black pigs, which were primarily haplotype A (denoted as G-T-C: nt 165901487 is G, nt 165901767 is T and nt 165902078 is C), similar to European pigs. Quantitative PCR data from liver tissues demonstrated that haplotype B individuals had higher CYB5A expression than did those with haplotype A. This was confirmed by in vitro cell transfection assays, in which haplotype B individuals had higher reporter activity than did those with haplotype A. In silico analysis predicted that Myc-associated zinc-finger protein (MAZ) is a potential transcription factor at position 165901767. Electrophoretic mobility shift assays showed this polymorphism affects the stable binding of transcription factors to the CYB5A promoter, which in turn affects the expression levels of this gene. Therefore, this variation of the porcine CYB5A promoter region may explain the differences in androstenone accumulation between Chinese and European pig breeds and may also prove useful as a genetic marker to distinguish the origin of different pig breeds.

Effect of ORF119 gene deletion on the replication and virulence of orf virus.

Orf is a severe infectious disease of sheep and goats caused by orf virus (ORFV). To investigate the role of ORF119 gene of ORFV, we constructed ORFV with deleted ORF119 gene and LacZ as reporter gene (ORFV-Δ119-LacZ) via homologous recombination. The results showed that wild-type ORF-SHZ1 and ORFV-Δ119-LacZ deletion viruses replicated in Vero cells to similar titers. Relative transcriptional levels of virulence genes OVIFNR, GIF, VEGF and VIL-10 of ORFV-Δ119-LacZ deletion virus were slightly but not significantly lower after 24 hr compared with the wtORF-SHZ1 virus. In vivo experiments showed that 2-month-old lambs inoculated with ORFV-Δ119-LacZ deletion virus exhibited a similar total clinical score compared with those inoculated with wtORF-SHZ1 virus. Based on these results, we conclude that deletion of the ORF119 gene has no significant effect on ORFV replication and virulence.

Alendronate, a bisphosphonate, increased upper and lower gastrointestinal bleeding: risk factor analysis from a nationwide population-based study.

UNLABELLED: Patients receiving alendronate for osteoporosis carry a significantly higher risk of developing upper gastrointestinal bleeding (GIB) and lower GIB (hazard ratio 1.32 and 1.84, respectively) after adjusting for potential confounding factors such as age, gender, co-morbidity, and some medications. The risk factors associated with GIB were further analyzed. INTRODUCTION: Patients receiving alendronate, a type of bisphosphonate, for osteoporosis have a higher risk of developing upper gastrointestinal bleeding (UGIB). Whether patients receiving alendronate also have a higher risk of lower gastrointestinal bleeding (LGIB) has not been studied. In this study, we investigated the association between GIB and alendronate use and to identify the possible risk factors of GIB among alendronate users. METHODS: Using the National Health Insurance (NHI) Research Database of Taiwan, 3,000 alendronate users and 12,000 age-, sex-, and enrollment time-matched controls were extracted for analysis from a cohort data set of 1,000,000 randomly sampled subjects. Cox proportional hazard regression models were used to identify the risk factors for UGIB and LGIB in all enrollees and alendronate users after adjustments for age, gender, comorbidity (hypertension, diabetes mellitus, coronary heart disease, heart failure, chronic renal disease, chronic obstructive pulmonary disease, peptic ulcer, and cirrhosis), and medications (nonsteroidal anti-inflammatory drugs [NSAIDs], aspirin, steroids, clopidogrel, ticlopidine, warfarin, and selective serotonin reuptake inhibitors). RESULTS: During a median of 1.30-year follow-up, patients receiving alendronate had significant higher risk of UGIB and LGIB after adjusting for age, gender, and potential confounding factors such as comorbidity and medications. Age, chronic renal disease, NSAID, and clopidogrel use may be independent risk factors for UGIB among alendronate users. Age, male gender, clopidogrel, and ticlopidine use may be independent risk factors for LGIB among alendronate users. CONCLUSION: Patients receiving alendronates seemed to carry a higher risk for UGIB and LGIB, respectively, after adjustment for age, sex, underlying comorbidity, and certain medications.

Causal association between complement system FHR-5, CTRP9, and breast carcinoma in situ: a Mendelian randomization study.

OBJECTIVE: Breast carcinoma in situ accounts for a significant number of newly diagnosed breast cancer cases. However, the cause of this type of cancer is unclear, which has led to debates regarding treatment strategies. A Mendelian randomization (MR) study was conducted to explore whether complement system or complement C1q/tumor necrosis factor-related proteins (CTRPs) are causally associated with breast carcinoma in situ. MATERIALS AND METHODS: This two-sample multivariable MR study used genome-wide association study (GWAS) data for all complement system factors and CTRPs. Summary-level statistics were obtained from the breast carcinoma in situ GWAS database. The study employed the MR-Egger method, inverse variance weighted (IVW) method, and weighted median method. Additionally, sensitivity analyses, including the MR-Egger intercept, funnel plot, and leave-one-out analysis, were conducted to address uncertainties and enhance the reliability of the findings. RESULTS: The study indicated that certain immunomodulatory molecules might increase the risk of breast carcinoma in situ, with consistent results. Specifically, CTRP9 showed a 57.0% increased risk [IVW: odds ratio (OR) 0.570 (0.350, 0.928), p < 0.05], and complement factor H (FH)-related protein 5 (FHR-5) was linked to a 67.2% higher risk [IVW: OR 0.672 (0.477, 0.947), p < 0.05]. However, no associations were found with other molecules, suggesting the relationship between immunomodulatory molecules and cancer may be context-specific. CONCLUSIONS: This MR study marks the initial identification of a direct link between FHR-5 and CTRP9 and the susceptibility to breast carcinoma in situ. Delving into the roles of immunomodulatory molecules and immune responses within the tumor microenvironment holds considerable importance for the management of breast carcinoma in situ.

Genetic divergence, range expansion and possible homoploid hybrid speciation among pine species in Northeast China.

Although homoploid hybrid speciation in plants is probably more common than previously realized, there are few well-documented cases of homoploid hybrid origin in conifers. We examined genetic divergence between two currently widespread pines in Northeast China, Pinus sylvestris var. mongolica and Pinus densiflora, and also whether two narrowly distributed pines in the same region, Pinus funebris and Pinus takahasii, might have originated from the two widespread species by homoploid hybrid speciation. Our results, based on population genetic analysis of chloroplast (cp), mitochondrial (mt) DNA, and nuclear gene sequence variation, showed that the two widespread species were divergent for both cp- and mtDNA variation, and also for haplotype variation at two of eight nuclear gene loci surveyed. Our analysis further indicated that P. sylvestris var. mongolica and P. densiflora remained allopatric during the most severe Quaternary glacial period that occurred in Northeast China, but subsequently exhibited rapid range expansions. P. funebris and P. takahasii, were found to contain a mixture of chlorotypes and nuclear haplotypes that distinguish P. sylvestris var. mongolica and P. densiflora, in support of the hypothesis that they possibly originated via homoploid hybrid speciation following secondary contact and hybridization between P. sylvestris var. mongolica and P. densiflora.

Time-intensity curve parameters in colorectal tumours measured using double contrast-enhanced ultrasound: correlations with tumour angiogenesis.

AIM: The aim of the study was to assess the correlation between time-intensity curve (TIC) parameters and colorectal tumour angiogenesis using double contrast-enhanced ultrasound (DCEUS), in which an intraluminal contrast agent was used in combination with an intravascular contrast agent. METHOD: Thirty nine patients with colorectal tumours were examined preoperatively. During hydrocolonal examination with the intraluminal contrast agent, an intravascular contrast agent, SonoVue, was used to perform the DCEUS. The parameter arrival time (AT), time to peak (TTP), peak intensity (PI) and area under the curve (AUC) were measured. Postoperative specimens were assessed for microvessel density (MVD) and vascular endothelial growth factor (VEGF). The correlation between TIC parameters and the expression of VEGF or MVD was studied. RESULTS: The mean values of AT, TTP, PI and AUC of the colorectal tumours were 14.32 ± 11.36 s, 30.61 ± 18.65 s, 20.38 ± 17.45 dB and 221.10 ± 156.09 dB.s, respectively. Both AUC and MVD were significantly higher in colorectal adenocarcinomas than in adenomas (all P < 0.05). A positive linear correlation was found between the AUC and MVD in colorectal tumours (r = 0.686, P = 0.0019). No correlation was found between VEGF and any TIC parameter. CONCLUSION: DCEUS is a valuable method for evaluating angiogenesis in colorectal tumours in vivo. The AUC has a positive linear correlation with MVD and could form a new index for assessing angiogenesis and the biological behaviour of colorectal tumours.

Species delimitation and biogeography of two fir species (Abies) in central China: cytoplasmic DNA variation.

It remains unclear how speciation history might contribute to species-specific variation and affect species delimitation. We examined concordance between cytoplasmic genetic variation and morphological taxonomy in two fir species, Abies chensiensis and A. fargesii, with overlapping distributions in central China. Range-wide genetic variation was investigated using mitochondrial (mt) and plastid (pt) DNA sequences, which contrast in their rates of gene flow. Four mtDNA haplotypes were recovered and showed no obvious species' bias in terms of relative frequency. In contrast, a high level of ptDNA variation was recorded in both species with 3 common ptDNA haplotypes shared between them and 21 rare ptDNA haplotypes specific to one or other species. We argue that the lack of concordance between morphological and molecular variation between the two fir species most likely reflects extensive ancestral polymorphism sharing for both forms of cytoplasmic DNA variation. It is feasible that a relatively fast mutation rate for ptDNA contributed to the production of many species-specific ptDNA haplotypes, which remained rare due to insufficient time passing for their spread and fixation in either species, despite high levels of intraspecific ptDNA gene flow. Our phylogeographic analyses further suggest that polymorphisms in both organelle genomes most likely originated during and following glacial intervals preceding the last glacial maximum, when species distributions became fragmented into several refugia and then expanded in range across central China.

First Report of Grapevine Trunk Disease Caused by Botryosphaeria obtusa in China.

In September 2010, grapevine (Vitis vinifera) trunk diseases were observed in several vineyards of Yantai District in Shandong Provinces and Changli County of Hebei Provinces of China. Characteristic symptoms of Botryosphaeria canker were apparent, including dark brown discoloration on the trunk (visible in cross-section), cob base shriveling, drying of fruit clusters, and berry falling (2). To identify the causal pathogen, culturing of fungi was attempted from 387 small pieces of tissue from the canker margins of 43 diseased plants. Samples were surface disinfected by placing them in 75% ethanol for 1 min and rinsing with sterilized water three times before culturing on potato dextrose agar (PDA) at 28°C for 7 to 10 days. Fungi isolated were single spored to obtain pure cultures. On the basis of colony characteristics on PDA, 18 isolates from the 387 tissue pieces were eventually identified as Botryosphaeria obtusa (1), Most of the other fungi isolated were B. dothidea. B. obtusa colonies were grayish white, becoming dark brown with age, and pycnidia were formed after incubation for approximately 7 days. Conidia measured 8 to 11 × 17 to 26 µm (n= 50). Two isolates were used for rDNA internal transcribed spacer (ITS) sequence analysis with primers ITS1 and ITS4 (3). PCR products were separated by electrophoresis and bands were purified for legation with PMD-18T (Takara Company, Dalian, China) vector for sequencing. BLAST searches of two ITS sequences had 99 to 100% identity to B. obtusa. EF1-α and ß-tubulin sequence analysis gave similar results. Koch's postulates were completed in the greenhouse on grape shoots inoculated with two isolates of B. obtusa originally isolated from diseased plants in the field. Inoculations were made on green shoots of V. vinifera cv. Dunkelfelder T. Six shoots were inoculated per isolate by wounding with a 4-mm cork borer (2 mm deep) and placing a colonized agar plug from a 5-day-old culture on the wound and wrapping it with Parafilm. Controls were mock inoculated with an agar plug from sterile PDA. Inoculated shoots were incubated in the dark under moist conditions in the laboratory for 8 to 10 days at 25°C. Inoculated shoots had necrotic cankers after 8 to 10 days and B. obtusa was recovered from each canker margin. The results indicated that some grapevines in China with symptoms of Botryosphaeria canker were infected by B. obtusa. To our knowledge, this is the first report of this pathogen causing trunk disease on grapevine in China. References: (1) A. Taylor et al. Australas. Plant Pathol. 34:187, 2005. (2) J. R. Úrbez-Torres et al. Plant Dis. 92:519, 2008. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

[Effects of positive end-expiratory pressure setting of mechanical ventilation guided by esophageal pressure in the treatment of patients with traumatic craniocerebral injury combined with acute respiratory distress syndrome].

Objective: To investigate the effects of positive end-expiratory pressure (PEEP) setting of mechanical ventilation guided by esophageal pressure in the treatment of patients with traumatic craniocerebral injury combined with acute respiratory distress syndrome (ARDS). Methods: The retrospective cohort study was conducted. From June 2016 to June 2018, 55 patients with traumatic craniocerebral injury combined with ARDS who met the inclusion criteria were admitted to Zhengzhou Central Hospital Affiliated to Zhengzhou University. According to PEEP setting method, 28 patients were allocated to esophageal pressure group (17 males and 11 females, aged (40±13) years) and 27 patients were allocated to PEEP-fractional concentration of inspired oxygen (FiO2) table group (18 males and 9 females, aged (38±10) years). Patients in the 2 groups were treated with mechanical ventilation guided by lung protective ventilation strategy, and the optimal PEEP at 0 (immediately), 24, 48, and 72 h after treatment was determined according to esophageal pressure and PEEP-FiO2 table, respectively. The mechanical ventilation parameters in the 2 groups were adjusted according to the optimal PEEP. The transpulmonary end-expiratory pressure, pulmonary compliance, oxygen index, central venous pressure, mean arterial pressure, and intracranial pressure at 24, 48, and 72 h after treatment were recorded. Data were statistically analyzed with analysis of variance for repeated measurement, chi-square test, independent sample t test, and Bonferroni correction. Results: The optimal PEEP of patients in esophageal pressure group at 0, 24, 48, and 72 h after treatment was (12.4±3.9), (11.2±3.5), (13.4±2.6), and (13.2±3.6) cmH2O (1 cmH2O=0.098 kPa), respectively, which was significantly higher than (8.2±2.5), (7.4±2.2), (8.3±2.3), and (8.5±2.5) cmH2O in PEEP-FiO2 table group, respectively (t=4.702, 4.743, 7.849, 5.623, P<0.01). The transpulmonary end-expiratory pressure and pulmonary compliance at 24, 48, and 72 h after treatment and oxygen index at 48 and 72 h after treatment of patients in esophageal pressure group were significantly higher than those in PEEP-FiO2 table group (t=17.852, 20.586, 19.532, 4.752, 5.256, 7.446, 2.342, 4.178, P<0.05 or P<0.01). The central venous pressure of patients in esophageal pressure group at 24, 48, and 72 h after treatment was significantly higher than that in PEEP-FiO2 table group (t=12.632, 5.247, 8.994, P<0.01), and there was no statistically significant difference in mean arterial pressure of patients between the 2 groups at 24, 48, and 72 h after treatment (P>0.05). The intracranial pressure of patients in esophageal pressure group was higher than that in PEEP-FiO2 table group at 24, 48, and 72 h after treatment, but there was no statistically significant difference between the 2 groups (P>0.05). Conclusions: For patients with traumatic craniocerebral injury combined with ARDS, the optimal PEEP can be set under the guidance of esophageal pressure method, and the mechanical ventilation parameters adjusted according to the optimal PEEP can improve lung compliance and accelerate recovery of lung function more effectively, with no adverse effect in mean arterial pressure and intracranial pressure.

First Detection of Virulence in Puccinia striiformis f. sp. tritici in China to Resistance Genes Yr24 (=Yr26) Present in Wheat Cultivar Chuanmai 42.

Stripe rust disease of wheat caused by Puccinia striiformis f. sp. tritici was observed on previously resistant bread wheat (Triticum aestivum L.) cv. Chuanmai 42 during the 2008-2009 crop season in Pi County, Sichuan Province, China. More than 10 single pustules were isolated from the diseased leaf samples collected in the field and inoculated on 7-day-old susceptible wheat seedlings cv. Mingxian 169. After 18 to 24 h of incubation at 100% relative humidity in darkness, the plants were moved into the greenhouse, maintained at 15 to 18°C, and supplemented with 10,000 lx of fluorescent light for 10 h per day. The second leaves were clipped when chlorotic spots appeared on leaves (~7 days postinoculation), and plants were covered with glass cylinders to prevent cross contamination. Urediniospores of each isolate were collected 16 days after inoculation and temporarily kept in a dryer at low temperature (3 to 4°C). The virulence spectra of the isolates were tested on Chinese differentials and wheat lines with known Yr genes at the seedling stage (1). A new Yr24 (=Yr26) virulent pathotype, different from currently known pathotypes in China, was identified. To our knowledge, this is the first detection of Yr24 virulence in Puccinia striiformis f. sp. tritici populations on Chuanmai 42. In addition, the new pathotype was also virulent to Lantian 17, Guinong 22 (Chinese differential), and 92R137 derived wheat lines Nannong 04Y10 and Nannong 05Y628, known to carry Yr24 (2,3). The avirulence/virulence formula of the new pathotype is Yr1, 3, 4, H46, 5, 6, 15, 17, 18, 32, Sp, Sd/Yr2, 8, 9, 10, 12, 24 (=26), 31, and Su. Wheat cultivars and breeding materials, previously protected by Yr24 gene, are now vulnerable to stripe rust epidemics in the region. Pure isolates of the new pathotype (Accession No. 09-6-16-3) are stored in the Chinese Academy of Agricultural Sciences (CAAS; Beijing) stripe rust collection. References: (1) W. Q. Chen et al. Plant Dis. 93:1093, 2009. (2) G. Q. Li et al. Theor. Appl. Genet. 112:1434, 2006. (3) Z. F. Li et al. Plant Dis. 90:1302, 2006.

Race Dynamics, Diversity, and Virulence Evolution in Puccinia striiformis f. sp. tritici, the Causal Agent of Wheat Stripe Rust in China from 2003 to 2007.

Stripe (or yellow) rust caused by Puccinia striiformis f. sp. tritici is the most destructive foliar disease of wheat in China. The pathogen populations were analyzed for virulence evolution, complexity, phenotypic dynamics, and diversity on temporal and spatial bases. A total of 41 races were identified and characterized from 4,714 stripe rust isolates collected during 2003 through 2007 from wheat growing areas in 15 provinces in China. The races were based on avirulence/virulence patterns to 19 differential host genotypes. Chinese stripe rust population exhibited high diversity with a complex virulence structure. Comparisons using the relative Shannon's index indicated that some differences in the richness and evenness of races were present in pathogen populations within years and between regions despite a national tendency to reduced diversity over time. A noticeably increased frequency of race CYR33 (Chinese yellow rust 33) with virulence for YrSu was the major virulence change recorded in this study compared to the results on an annual basis. Isolates of Puccinia striiformis f. sp. tritici from different regions showed differences in the composition of races, distribution frequency, and diversity. The uneven distribution of major races and comparatively greater diversity in the Northwest and Southwest regions than that in the Huang-Huai-Hai region suggest that long-distance migrations of the pathogen occur from one or more over-summering areas eastward into over-wintering areas. This supports the hypothesis that southern Gansu and northwestern Sichuan comprises a "center of origin for virulence". Mutation of virulence or avirulence for host resistance in the stripe rust fungus may be the basic cause of the occurrence of new virulent types. The subsequent dominance of certain races will vary with parasitic fitness and the opportunities to be selected through large-scale cultivation of varieties with matching resistance genes. Implications of the center of origin for virulence variation and diversity in the pathogen population and an alternative strategy for limiting virulence evolution are discussed.

Incubation of excised apothecia enhances ascus maturation of Sclerotinia sclerotiorum.

Synchronized maturation of ascospores of Sclerotinia sclerotiorum is desirable for establishing a transformation system, conducting genetic analyses of the pathogen, defining the precise epidemiological roles of ascospores and screening plant germplasm for resistance. In general, fresh apothecia collected from germinated sclerotia contained primarily immature or discharged asci. This study was undertaken to investigate whether maturation of asci and ascospores could be enhanced by incubation of excised apothecia and to determine the effects of factors such as temperature, excision time, light and ventilation on maturation of asci and ascospores in excised apothecia. Maturation of asci was compared between intact and excised apothecia that were incubated under similar conditions. Results demonstrated that temperature was an important factor affecting ascus maturation of S. sclerotiorum during incubation of excised apothecia, and the optimum temperature was around 21 C. After incubation at 21 C for 30 h, the percentage of undischarged mature asci in excised apothecia increased up to 70-80%. This increase was accompanied by a significant increase in ascospore production of up to 5 x 10(5) ascospores per apothecium. Detailed time course studies indicated that mature asci peaked at 30-36 h of postexcision incubation. Mature asci and the number of ascospores were higher in open incubation than in closed incubation, suggesting that accumulation of volatile substances was not required for ascus/ascospore maturation during postexcision incubation and ventilation could enhance the maturation process. Light also did not affect the maturation of asci during the incubation of excised apothecia. Germination rates for ascospores from excised apothecia under various treatments were similar to those from untreated apothecia but declined slightly with time postexcision. The incubation of excised apothecia promoted ascus maturation compared with intact apothecia.