[Metabolic study of iron deposition based on magnetic resonance in patients with nonalcoholic fatty liver disease].

Objective: To explore the relationship between liver iron deposition and steatosis in patients with non-alcoholic fatty liver disease (NAFLD) through MRI. Methods: 163 cases of liver biopsy underwent MRI examination. R2* was used to measure liver iron content. Dixon-based proton density fat fraction (PDFF) was used to measure liver fat content. One-way ANOVA, r-correlation, ROC curve, and others were used to assess the relationship between clinical case data, serological indices, and imaging results in accordance with the pathological results of the liver biopsy. Results: R2* gradually increased as the pathological steatosis grade rose. The R2* that corresponded to no steatosis (< 5%), mild steatosis (14.95%±8.55%), moderate steatosis (46.30%±9.32%), and severe steatosis (73.86%±6.35%) were 27.56±4.40, 31.06±5.95, 38.06±4.80, and 48.10±5.55 (P < 0.001), respectively. There was a positive correlation between R2* and liver steatosis content (r= 0.769, P < 0.05). The area under the ROC curve and cut-off value were 0.88 and 31.77, respectively, and there was no distinct relationship with liver inflammation or fibrosis. Conclusion: R2* can quantitatively and non-invasively evaluate liver iron deposition in patients with NAFLD. A distinct relationship exists between liver steatosis and iron deposition, and iron deposition tends to increase as the steatosis aggravates.

[Effects of cinepazide maleate injection on blood pressure in patients with acute ischemic stroke and hypertension].

Objective: To investigate the blood pressure change in patients with acute ischemic stroke (AIS) and hypertension treated with cinepazide maleate injection. Methods: This was a subgroup analysis of post-marketing clinical confirmation study of cinepazide maleate injection for acute ischemic stroke: a randomized, double-blinded, multicenter, placebo-parallel controlled trial, which conducted in China from August 2016 to February 2019. Eligible patients fulfilled the inclusive criteria of acute anterior circulation ischemic stroke with National Institutes of Health Stroke Scale (NIHSS) scores of 7-25. The primary endpoints were mean blood pressure of AIS patients treated with cinepazide maleate or control, which were assessed during the treatment period (14 days), and the proportion of the patients with normal blood pressure was analyzed after the treatment period. Furthermore, a subgroup analysis was performed to investigate a possible effect of the history of hypertension on outcomes. Results: This analysis included 809 patients with hypertension. There was no significant difference in patients blood pressure and the proportion of patients with normal blood pressure (60.5% vs. 59.0%,P>0.05) between cinepazide maleate group and control group. Conclusion: Administration of cinepazide maleate injection does not affect the management of clinical blood pressure in patients with AIS.

[Magnetic resonance imaging analysis on the effect of marathon on the dynamic changes of morphological characteristics of patellar tendon in amateur marathon runners after half marathon].

Objective: To explore the dynamic changes of morphological characteristics of patellar tendon (PT) in amateur athletes after a half-marathon using magnetic resonance imaging. Methods: A total of 19 male amateur marathon runners with 38 knee joints,aged from 26 to 53(41.5±7.9) years, running for 3 to 18 years, with a weekly running volume of 30-90 km and a monthly running volume of 100-300 km were enrolled and underwent 1.5T MRI scan before the half-marathon, within 3 hours after running and 3 days after running. Ten healthy male volunteers with 20 knee joints, who had never participated in marathon and exercised (including but not limited to running) per week for less than 150 minutes were recruited as the control group, aged from 26 to 54 (39.4±9.1) years. Firstly, the PT signal was qualitatively assessed on fat-suppressed proton density-weighted imaging (fs-PDWI) sequence to observe the presence of patellar tendinitis. Then, the length, proximal, middle and distal cross-sectional area (CSA), and volume of PT were measured using the post-processing tool ITK-SNAP, and the data were standardized. The independent sample t-test was used for comparing. One-way repeated measures analysis of variance was used to analyze the morphological changes of PT before and after half-marathon running. Results: The incidence of asymptomatic patellar tendinitis in amateur marathon runners was 26.3% (5/19). No significant MR signal changes of PT were observed in all runners after running. The proximal CSA in runners group was larger than that in controls [(4.20±0.62) mm2/kg3/4 vs (3.63±0.57) mm2/kg3/4, P<0.05], and there was no significant difference in length, medium and distal CSA and volume(all P>0.05). The length, proximal and distal CSA and volume of PT in runners group increased at 3 h after running [(47.35±3.22) mm vs (46.83±3.35) mm; (102.52±13.03) mm2 vs (98.98±13.14) mm2; (108.67±15.72) mm2 vs (100.27±14.37) mm2; (4 020.36±514.38) mm3 vs (3 826.57±499.23) mm3, all P<0.05]. There was no significant difference between before running and 3 days after running(all P>0.05). The middle CSA were not significantly different among different periods(all P>0.05). Conclusion: Marathon has effect on the normal PT morphology in male amateur marathon runners, showing an increase in proximal CSA. A half-marathon will cause reversible changes in PT length, regional CSA and volume.

[Diffusion tensor imaging in quantitative evaluation on thigh muscle of male amateur marathon runners after running a half marathon].

Objective: To investigate the effect of half marathon on thigh muscle in male amateur marathon runners by diffusion tensor imaging (DTI). Methods: A total of 17 male amateur marathon runners aged from 27 to 57 (43.7±2.8) years were recruited from May to August 2020 in Hangzhou, Zhejiang Province. MRI examination of bilateral thigh muscles were performed before and 3 h, 3 d and 7 d after a half marathon. The fractional anisotropy (FA) values was obtained by DTI sequence. The FA values of rectus femoris, intermedius femoris, medial femoris, lateral femoris, biceps femoris longus, semitendinosus, semimembranosus, adductor magnus and adductor longus were measured on the horizontal axis of bilateral thigh muscles. Friedman M test was used to compare the changes of FA values of each muscle at each time point before and after running, and pairwise comparison of FA values of statistically significant muscles at 3 h, 3 d and 7 d after running was performed. Results: The overall FA value of thigh muscle group [M (Q1, Q3)] at 3 h after running was decreased compared to before running [0.24 (0.20, 0.28) to 0.25 (0.21, 0.29), P<0.001], and there was no significant difference between baseline values at 3 d and 7 d after running (all P>0.05). FA values of vastus intermedius, vastus medialis, semimembranosus and adductor magnus at 3 h after running were lower than those before running [(0.19 (0.18, 0.22) vs 0.21 (0.19, 0.24), 0.19 (0.17, 0.20) vs 0.21 (0.18, 0.23), 0.26 (0.24), 0.29) vs 0.27 (0.15, 0.30) and 0.20 (0.19, 0.22) vs 0.21 (0.20, 0.23), both P<0.05], and there was no statistical significance between 3 d and 7 d after running and those before running (all P>0.05). FA value of vastus lateralis muscle at 3 h after running decreased compared with that before running, but the difference was not statistically significant (P>0.05). FA value began to increase at 7 d after running, and the difference was statistically significant [0.24 (0.21, 0.27) vs 0.23 (0.19, 0.25), P = 0.002]. FA value of rectus femoris muscle at 3 h after running decreased compared with that before running, but the difference was not statistically significant (P>0.05), and began to increase at 3 d after running and the difference was statistically significant [0.29 (0.26, 0.34) vs 0.26 (0.23, 0.29), P=0.006]. FA value of adductor longus muscle increased at 3 h after running, but the difference was not statistically significant (P>0.05). FA value continued to increase at 3 d and 7 d after running, and the difference was statistically significant [0.23 (0.21, 0.25) vs 0.22 (0.19, 0.24), 0.23 (0.21, 0.26) vs 0.22 (0.19, 0.24), all P<0.05]. Conclusions: The change of FA value of thigh muscle after half marathon is reversible. At 3 h after half marathon, FA values of femoris intermedius, femoris medialis, semimembranosus muscle and adductor magnus muscle of amateur marathon runners decreased most obviously, which may be the dominant muscle group during running.

[Study of liver fat and iron deposition quantification based on magnetic resonance imaging in rats with nonalcoholic fatty liver disease].

Objective: To investigate the accuracy of magnetic resonance imaging (MRI) for quantitative determination of liver fat and iron content through a rat model of non-alcoholic fatty liver disease (NAFLD) induced by methionine-choline deficient (MCD) diet. Methods: Sixty SD rats were randomly divided into experimental (MCD-diet group, n = 30) and normal control group (normal diet, n = 30). Rats were subjected to special MRI examinations at the ends of 2, 4, and 8 weeks. Proton density fat fraction (PDFF) and R2* value were obtained, and then the rats were sacrificed. The liver tissues were stained with HE, Prussian blue, etc. Liver tissue non-heme iron (NHI) homogenate was determined by flame atomic absorption spectrometry. According to different data, one-way analysis of variance, t-test or χ (2) test was used for statistical analysis. Results: PDFF and R2 * values in the MCD diet group at 2, 4 and 8 weeks were 23.37% ± 9.20%, 28.07% ± 6.84%, 25.40% ± 7.04% (P < 0.01) and 90.58 ± 15.92, 104.12 ± 13.47, 106.35 ± 15.76 (P < 0.05), respectively, which were significantly higher than the normal control group PDFF (2.39% ± 0.50%, 2.45% ± 0.45%, 3.26% ± 0.80%) and R2* (48.93 ± 7.90, 54.71 ± 5.91, 64.25 ± 15.76). Additionally, with the disease progression, R2 * had gradually increased, which was consistent with the NHI trend in liver tissue homogenates of each group. Conclusion: MRI, as a non-invasive quantitative method, can accurately assess liver fat and iron content in fatty liver disease, and with the degree of severity of fat changes, iron deposits tend to increase.

[Studying the correlation between ferritin and non-alcoholic fatty liver disease].

Objective: To analyze the correlation between serum ferritin and steatosis in non-alcoholic fatty liver disease. Methods: Data of 167 cases who underwent liver biopsy in the Affiliated Hospital of Hangzhou Normal University were collected. Hydrogen proton magnetic resonance spectroscopy were performed within one week. The pathological results of liver biopsy were used as the gold standard to analyze the case data, serological indicators, magnetic resonance spectroscopy-proton density fat fraction. Results: Pathological monitoring result showed that the serum ferritin in patients without steatosis, and with mild, moderate and severe steatosis were (206.20 ± 189.83), (286.65 ± 200.80), (326.55 ± 214.71), (391.50 ± 184.93) ng/ml, respectively, P < 0.005. Serum ferritin was correlated to body mass index, PDFF, alanine aminotransferase, gamma glutamyltransferase, low-density lipoprotein, high-density lipoprotein. The area under ​​the receiver operating characteristic curve with ferritin for the diagnosis of non-alcoholic fatty liver disease was 0.716, and the optimal diagnostic threshold was 214.56 ng/ml. The sensitivity and specificity were 80.1%, and 68.8%, respectively. There was no statistically significant difference between the intralobular inflammation, fibrosis, and ferritin. Prussian blue iron staining had no apparent deposition of iron particles. Conclusion: Ferritin has significant positive correlation with the results of pathological and magnetic resonance imaging for liver steatosis. Therefore, it can be used as a non-invasive diagnostic method for liver steatosis evaluation.

[A preliminary study of resting-state fMRI in patients with diarrhea-type irritable bowel syndrome after pinaverium bromide combined treatment with flupentixol-melitracen].

Objective: To observe the difference of brain activity in patients with diarrhea-type irritable bowel syndrome (IBS-D) treated with pinaverium bromide (PB) combined with flupentixol-melitracen (FM), and to explore the mechanism of efficacy of combined with anxiolytic/antidepressant drugs in IBS-D patients at the central level, using resting-state functional magnetic resonance imaging (rs-fMRI). Methods: Forty-eight patients with IBS-D, including 28 males, 20 females, mean age 22-48 (33±7) years, were selected from the Affiliated Hospital of Hangzhou Normal University from October 2015 to October 2018.All patients with IBS-D underwent rs-fMRI scans before and after receiving either PB (basic treatment group, n=16), PB combined with FM (combination therapy group, n=16), or no medication (no treatment group, n=16). Rs-fMRI regional homogeneity (ReHo) parameter among the three groups of patients were compared using One-way ANOVA analysis and post analysis.Partial correlation and mediation analyses were performed on ReHo values and the improvement of symptoms scores (gastrointestinal symptom rating scale(GSRS) and hospital anxiety/depression scale (HAD)) in the two medicated groups. Results: No significant differences in ReHo values were observed among the three groups before treatment. Compared with patients on no-medication, patients receiving either PB or PB-FM showed decreased ReHo in the striatum, insular lobe, medial prefrontal cortex (MPFC) and subcallosal gyrus, and increased ReHo in the occipital cortex. In particular, the combined treatment group showed more extensive decreased ReHo in the left thalamus and left temporal pole, and increased in the left precuneus. Compared with the basic treatment group, the combined treatment group showed decreased ReHo in the right putamen, right insula, right MPFC and subcallosal gyrus, and increased ReHo in the left precuneus. In addition, the combined treatment group demonstrated a positive correlation between ReHo values in the left thalamus and the improvement of HAD score (r=0.653, P=0.011) , and a negative correlation between ReHo values in left precuneus and the improvement of GSRS and HAD score (r=-0.771, P=0.001; r=-0.716, P=0.004). ReHo values in the left precuneus were observed to mediate between gastrointestinal symptoms and anxiety-depressive symptoms in the combined treatment group. Conclusions: The efficacy of PB combined with FM is superior to PB alone in the treatment of IBS. In addition to more extensive changes in pain-related brain areas, IBS-D patients treated with anxiolytic/antidepressant also show changes in default network and brain areas related to emotional regulation, and are associated with improvement in gastrointestinal symptoms, anxiety and depression.

[Study on diffusion kurtosis imaging in hyperacute phase of mild traumatic brain injury].

Objective: To explore the changes of cerebral white matter in the hyperacute period (<24 h) patients with mild traumatic brain injury (mTBI) by diffusion kurtosis imaging(DKI) technique. Methods: A total of 52 patients with mTBI were included in this study, collected in Emergency Department of Affiliated Hospital of Hangzhou Normal University from May 2016 to June 2017. Twenty-one healthy controls were recruited at the mean time(gender, age and years of education were matched with the patients). DKI data were acquired with 3.0 T scanners.The FSL software was used to preprocess the DKI data, and the white matter abnormalities were detected by tract-based spatial statistics (TBSS). Results: There were no statistical differences in gender, age and years of education between mTBI patients and healthy controls (P=0.427, P=0.235, P=0.165). The values of MK of the body of corpus callosum (BCC), the genu of corpus callosum (GCC), the splenium of corpus callosum (SCC), the bilateral anterior limb of interbal capsule (ALIP), the right posterior limb of internal capsule (PLIC_R), the bilateral anterior corona radiate (ACR), the bilateral posterior corona radiate (PCR), the bilateral superior corona radiate (SCR), the left inferior fronto-occipital fasciculus (IFOF_L)and the bilateral superior longitudinal fasciculus (SLF) were lower in mTBI patients((1.095±0.080), (1.130±0.066), (1.160±0.080), (1.135±0.077), (1.108±0.076), (1.203±0.069), (1.073±0.056), (1.052±0.055), (1.170±0.055), (1.149±0.050), (1.028±0.056), (1.051±0.059), (0.868±0.060), (1.194±0.048), (1.183±0.054) mm(2)/s) than those in healthy controls((1.153±0.054), (1.184±0.057), (1.215±0.068), (1.181±0.053), (1.163±0.062), (1.258±0.041), (1.115±0.037), (1.096±0.049), (1.210±0.040), (1.190±0.049), (1.063±0.042), (1.087±0.057), (0.913±0.063), (1.236±0.047), (1.214±0.038) mm(2)/s)(P<0.01). However, there were no statistical differences in fractional anisotropy (FA) between groups (P>0.01). Conclusion: DKI technology is more sensitive in detecting cerebral white matter abnormalities in patients with hyperacute mTBI which routine MRI findings was normal.

[Regional homogeneity in the patients of irritable bowel syndrome complicated with depression: a resting-state functional magnetic resonance imaging study].

Objective: To explore the changes of the brain regional homogeneity (ReHo) in the patients of irritable bowel syndrome (IBS) complicated with depression by using resting-state functional magnetic resonance imaging (rs-fMRI) and to investigate the relation between abnormal brain function and depression in those patients. Methods: A total of 35 patients with IBS were included in this study, collected in Gastroenterology Department of Affiliated Hospital of Hangzhou Normal University from June 2015 to June 2017.According to Hamilton's Depression Scale (HAMD)-17 score, the IBS patients was divided into two groups: IBS with depression group, 21 cases and non-depression group, 14 cases.Thirty-six healthy controls were recruited at the mean time (gender, age and years of education were matched with the patients). Rs-fMRI and 3D T1 data were acquired with 3.0 T scanners.For all subjects, ReHo, a method measuring the synchronism of regional spontaneous activities, was calculated and one-way ANOVA was performed to assess the group effects.For those regions with significant group effects, correlation between ReHo values and GSRS and HAMD scores was analyzed too. Results: Significant group effects were found in multiple regions: the right pons, the left inferior temporal gyrus (ITG)/fusiform, the left superior temporal gyrus (STG)/temporal pole, the right inferior frontal gyrus (IFG), the right middle occipital gyrus (MOG), the right caudate nucleus, the left posterior cingulate cortex/precuneus (pCC/pC), the left postcentral gyrus, the right inferior parietal lobule (IPF). Compared with control group, all the two patient groups had increased ReHo values in the left pCC/pC, the right IPF, the left postcentral gyrus and the left STG/temporal pole; all the two patient groups had decreased ReHo values in the right MOG and the right IFG.Compared with non-depression group, depression group had increased ReHo values in the right caudate nucleus, and decreased ReHo values in the right pons, the left pCC/pC, the right IPF, the left postcentral gyrus.Particularly, there was no significant difference in the ReHo value in the right caudate nucleus between the control group and the non-depressed IBS group.In the depression group, significantly positive correlations were observed between ReHo values and HAMD scores in the left pCC/pC (r=0.599, P=0.011), and significantly positive correlations were observed between ReHo values and GSRS in the left postcentral gyrus (r=0.547, P=0.023). Conclusions: IBS patients have abnormal ReHo regions in default network and pain regulation-associated brain area.IBS with depressed patients has its own characteristics in the brain regions which related with pain perception and self-regulation.IBS could has an abnormal self-regulation of visceral pain, which is associated with the occurrence of depression.Depression can aggravate the disorders of pain perception and other functions of IBS patients.

[Study of susceptibility weighted imaging on MR and pathologic findings to distinguish benign or malignant soft tissue tumor].

Objective: To explore the diagnostic performance of susceptibility weighted imaging (SWI)in distinguishing benign or malignant soft tissue tumor, and to study pathological observation. Methods: Sixty-eight patients with soft tissue tumor, who received no previous treatment or invasive examination, received routine preoperative MRI examination and SWI scanning. The graduation and distribution of intratumoral susceptibility signal intensity(ITSS) and proportion of tumor volume were observed.The pathological results were also included for comparative analysis. Results: Fourty of 68 patients were benign and 28 were malignant. 72.5% (29/40) patients with benign soft tissue tumors were ITSS grade 1 and ITSS grade 3 (hemangioma). 89.3%(25/28) patients with malignant soft tissue tumors were ITSS grade 2 and ITSS grade 3. The difference was statistically significant (P<0.01). The distribution of ITSS in patients with benign soft tissue tumors was dominated by peripheral distribution and diffuse distribution (hemangioma), accounting for 90.0% (36/40). The distribution of ITSS in patients with malignant soft tissue tumors mainly distributed in the central region, accounting for 78.6% (22 /28). The difference was statistically significant (P<0.01). The proportion of tumor volume occupied by ITSS in benign soft tissue tumors was <1/3 and> 2/3 (hemangioma), accounting for 90.0% (36/40). The volume of malignant soft tissue tumors were predominantly <1/3 , accounting for 82.1% (23/28). The difference was statistically significant (P<0.01). Conclusion: SWI is sensitive in displaying the vein and blood metabolites in soft tissue lesions, which is helpful for the differential diagnosis of benign and malignant tumors in soft tissue.

[Value of hydrogen proton magnetic resonance spectroscopy in the determination of liver triglyceride in patients with fatty liver disease and its influencing factors].

Objective: To investigate the value of 1H-magnetic resonance spectroscopy ((1)H-MRS) in determining the content of liver triglyceride in patients with fatty liver disease (FLD), as well as its influencing factors. Methods: A total of 124 patients with nonalcoholic fatty liver disease (NAFLD), chronic hepatitis B (CHB), or hepatitis B complicated by FLD who underwent liver biopsy in the Affiliated Hospital of Hangzhou Normal University were enrolled, and the clinical data, serological markers, FibroScan results, and (1)H-MRS results were collected. A correlation analysis was performed with the results of liver biopsy as the gold standard, and the influence of factors including hepatitic B virus (HBV) infection and obesity on accuracy was analyzed. A one-way analysis of variance was used for comparison of means between the three groups, and the LSD or SNK test (for homogeneity of variance) or the Tamhane's or Dunnett's test (heterogeneity of variance) was used for comparison between any two groups. The t-test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data. The MRS-PDFF receiver operating characteristic (ROC) curve was plotted, the area under the ROC curve (AUC) was calculated, the optimal cut-off points for the diagnosis of NAFLD were estimated, and sensitivity and specificity were calculated. Results: The NAFLD group (42 patients) and the CHB + NAFLD group (40 patients) had a significantly higher proton density fat fraction (PDFF, the content of triglyceride in the liver) than the CHB group (42 patients) (16.84±9.76/9.39 ± 5.50 vs 3.45 ± 1.63, P < 0.001). The results were significantly correlated with the degree of steatosis confirmed by liver biopsy (P < 0.001), but it was not significantly correlated with inflammation or fibrosis grade. The correlation analysis showed that the MRS-PDFF value measured by 1H-MRS was significantly correlated with body mass index (BMI), blood lipids, alkaline phosphatase, and blood glucose, while it was not significantly correlated with age, sex, or the presence or absence of hepatitis B. The ROC curve analysis showed that the AUCs of PDFF measured by 1H-MRS were 0.93, 0.974, and 0.976, respectively, for the diagnosis of steatosis S1(≥5%), S2(≥34%), and S3(≥66%), and the corresponding optimal thresholds were 5.14%, 11.16%, and 16.7%, respectively. Conclusion: 1H-MRS has a high diagnostic value in quantitative evaluation of the degree of liver steatosis in patients with FLD and is not affected by the factors such as HBV infection, age, and sex, while it is correlated with BMI and lipid metabolism.

[Clinical, pathological and imaging features of primary pelvic Ewing's sarcoma].

OBJECTIVE: To explore the clinical, pathological and imaging features of Ewing's sarcoma in pelvis and to improve knowledge and diagnosis of the disease. METHODS: A retrospective analysis of the clinical, pathological and imaging data of pathologically confirmed 13 cases of Ewing's sarcoma in pelvis was carried out between May 2008 and March 2016 in the Affiliated Hospital of Hangzhou Normal University, the Third Hospital of Hebei Medical University and the Second Hospital of Hebei Medical University. RESULTS: The median age 13 cases of pelvic primary Ewing's sarcoma was 17 years old.The X-ray and CT imagings showed osteolytic and mixed bone destruction, CT showed mixed type in 10 cases, 8 cases of bone tumors as a flocculent, 10 cases of bone expansion failure, 10 cases of periosteal reaction, the layered 5 cases, radial in 5 cases.Thirteen cases showed soft tissue mass, soft tissue mass was equal or slightly lower density.Four cases showed heterogeneous contrast enhancement.The lesions showed low signal in T1WI and mixed high signal in T2WI of magnetic resonance imaging(MRI). The boundary of the lesions were obscure, and 5 cases had patchy necrosis area, and 9 cases had incomplete false capsule, surrounding soft tissue was violated.Four cases showed heterogeneous contrast enhancement after MRI enhancement scan. CONCLUSIONS: The age of onset of Ewing's sarcoma of the pelvis is more concentrated in about 15 years.The imaging feaures are mixed bone destruction and more bone is swelling and permeability damage, soft tissue mass is larger, bone tumor is cloudy or acicular, periosteal reaction in a layered and radial, most cases show that the false envelope is not complete.Combined with clinical and imaging examination, the diagnosis of the disease can be made.

Identification of complete linkage disequilibrium in the DSG4 gene and its association with wool length and crimp in Chinese indigenous sheep.

The desmoglein 4 (DSG4) gene is a potential candidate in the search for genes that may affect wool traits, because of its function. This study aimed to screen for polymorphisms in partial exon 16 and 3ꞌUTR of the sheep desmoglein 4 DSG4 gene, and to test its possible association with wool length and crimp associated with fur. Overall, 326 sheep were scanned via single-strand conformational polymorphism assay, through three pairs of primers. The breeds included Tan, Han, and TanxHan from China, Polled Dorset from Australia, and Suffolk from Britain genotypes AA, BB, and AB for primer2 and genotypes DD, EE, and DE for primer3 were detected in native breeds. Six SNPs and 3-bp insertion/deletions were found in exon 16, of which 4 lead to amino acid substitutions. In addition, 1 SNP was found in 3ꞌUTR. The DSG4 genotype was found to be strongly associated with all wool traits that were considered in this study (P < 0.01). Sheep with the genotype MM had a higher least square mean compared to sheep with the genotype WW or WM with respect to birth scapular wool length (P < 0.01), crimp number of birth scapular wool crimp (P < 0.01), crimp number of weaning scapular wool crimp (P < 0.01), and crimp number of weaning rump wool crimp (P < 0.01, P < 0.05). In conclusion, our study is the first to demonstrate that the DSG4 gene may be a candidate, or major gene, which influences important wool traits.

Characterization of microRNAs from goat (Capra hircus) by Solexa deep-sequencing technology.

MicroRNAs (miRNAs) are an important class of small noncoding RNAs that are highly conserved in plants and animals. Many miRNAs are known to mediate a myriad of cell processes, including proliferation and differentiation, via the regulation of some transcription and signaling factors, which are closely related to muscle development and disease. In this study, small RNA cDNA libraries of Boer goats were constructed. In addition, we obtained the goat muscle miRNAs by using Solexa deep-sequencing technology and analyzed these miRNA characteristics by combining it with the bioinformatics technology. Based on Solexa sequencing and bioinformatics analysis, 562 species-conserved and 5 goat genome-specific miRNAs were identified, 322 of which exceeded 100 in the expression levels. The results of real-time quantitative polymerase chain reaction from 8 randomly selected miRNAs showed that the 8 miRNAs were expressed in goat muscle, and the expression patterns were consistent with the Solexa sequencing results. The identification and characterization of miRNAs in goat muscle provide important information on the role of miRNA regulation in muscle growth and development. These data will help to facilitate studies on the regulatory roles played by miRNAs during goat growth and development.

Polymorphisms of the myostatin gene (MSTN) and its relationship with growth traits in goat breeds.

Mutations in the myostatin (MSTN) gene can inactivate its expression and result in a non-functional protein, which leads to dramatic muscularity and a "double-muscling" phenomenon in many species. Using gene sequencing and polymerase chain reaction-single-strand conformation polymorphism methods, polymorphisms of the MSTN gene were investigated as a candidate marker for growth in 288 goats. The results showed 2 novel single nucleotide polymorphisms: DQ167575 g.197G>A and 345A>T. Three potential genotypes (AA, AB, and BB) of substitution 197G>A in the 5'-untranslated region were detected in the 2 breeds. The polymorphism (CC and CD) of substitution 345A>T in exon I was segregated. The genetic diversity analysis revealed that Boer goat and Anhui white goat possessed intermediate genetic diversity in the P1 and P3 loci. Significant associations between the genotypes of the P3 locus and body weight, body length, and body height were observed in Boer goat and Anhui white goat (P < 0.05). It could be inferred that the MSTN gene may be a major gene or linked to the major gene affecting the goat growth traits. The polymorphic site could be a molecular marker-assisted selection program for body weight.

First Report of Powdery Mildew Caused by Podosphaera fusca on Herba eupatorii in China.

Herba eupatorii, one of the most important Chinese medicinal herbs, belongs to the Asteraceae family. In June 2012, a previously unknown disease, tentatively identified as powdery mildew, was observed on H. eupatorii growing in Shangqiu, in eastern Henan Province, China. Symptoms began as white mycelium partially covering upper leaf surfaces; as the disease progressed, it spread to cover entire leaf surfaces. The infected leaves became yellow and necrotic at advanced stages of infection. Specimens consisting of infected leaves were maintained at the Plant-Microbe Interaction Laboratory at Shangqiu Normal University. Microscopic observations of the morphology of the fungus revealed oval primary conidia measuring 18 to 27 × 15 to 22 µm. A long unbranched germ tube that germinated laterally from the ends of conidia was observed in some samples. Conidiophores were cylindrical, simple unbranched, and composed of a basal cell with a swollen base and three to six barrel-shaped conidia formed in chains, measuring 112 to 180 × 9 to 12 µm. Mycelial appressoria were nipple-shaped. Chasmothecia were not observed in the collected samples. To verify the identity of the fungus, the internal transcribed spacer (ITS) rDNA was amplified with ITS1 and ITS4 primers (3) and sequenced. The sequences were deposited as GenBank Accession No. JX546297. Comparison with sequences in the GenBank database revealed that the ITS sequence was 100% homologous with the sequence of Podosphaera fusca on Calendula officinalis (AB525914) (2) and Syneilesis palmata (AB040349) (1). The ITS sequence analysis verified that the causal agent was P. fusca, which is reported to be a cosmopolitan powdery mildew fungus, parasitic on numerous plant species in the Asteraceae family. Koch's postulates were completed by inoculating healthy H. eupatorii plants with a conidial suspension (prepared in distilled water) of 105 conidia/ml collected from infected plants. Five plants were sprayed until the suspension ran off the leaves, while five additional plants were sprayed with distilled water as a control. Plants were maintained in a climate cell under the following conditions: day, 24°C, 16 h; night, 20°C, 8 h; 85% humidity. After 10 days, inoculated plants developed symptoms similar to those observed in the field, whereas control plants remained healthy. Further examination showed that the inoculated plants were infected by P. fusca. To our knowledge, this is the first report of P. fusca affecting H. eupatorii in China. Because there are no fungicides labeled for use on this plant, the appearance of powdery mildew caused by P. fusca could result in substantial production loss of H. eupatorii. References: (1) T. Hirata et al. Can. J. Bot. 78:1521, 2000. (2) S. Takamatsu et al. Persoonia 24:38, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Genetic differentiation of chinese indigenous meat goats ascertained using microsatellite information.

To investigate the genetic diversity of seven Chinese indigenous meat goat breeds (Tibet goat, Guizhou white goat, Shannan white goat, Yichang white goat, Matou goat, Changjiangsanjiaozhou white goat and Anhui white goat), explain their genetic relationship and assess their integrity and degree of admixture, 302 individuals from these breeds and 42 Boer goats introduced from Africa as reference samples were genotyped for 11 microsatellite markers. Results indicated that the genetic diversity of Chinese indigenous meat goats was rich. The mean heterozygosity and the mean allelic richness (AR) for the 8 goat breeds varied from 0.697 to 0.738 and 6.21 to 7.35, respectively. Structure analysis showed that Tibet goat breed was genetically distinct and was the first to separate and the other Chinese goats were then divided into two sub-clusters: Shannan white goat and Yichang white goat in one cluster; and Guizhou white goat, Matou goat, Changjiangsanjiaozhou white goat and Anhui white goat in the other cluster. This grouping pattern was further supported by clustering analysis and Principal component analysis. These results may provide a scientific basis for the characteristization, conservation and utilization of Chinese meat goats.

Effects of L-ascorbic acid, alpha-tocopherol and co-culture on in vitro developmental potential of porcine cumulus cells free oocytes.

The maturation and developmental potential on cumulus-cell-free oocytes is of great importance theoretically and practically. The present study was to investigate the effects of l-ascorbic acid, alpha-tocopherol and co-culture on in vitro developmental potential of porcine denuded oocytes (DOs). Porcine DOs were cultured in maturation medium supplemented with vitamin C (0, 50, 100, 250, 500, 750 microM) and vitamin E (0, 10, 20, 50, 100, 250 microm), respectively. And they were also co-cultured with dispersed cumulus cells (group CCscoculture), intact cumulus cells oocyte complexes (COCs) (group COCscoculture), and COCs whose oocytes were removed (group OOXcoculture), respectively. After 44 h incubation, the maturation rates, cleavage rates and blastocyst rates after parthenogenetic activation in three experiments mentioned above were collected and analysed, respectively. L-Ascorbic acid promoted porcine DOs in vitro maturation and blastocyt development after parthenogenetic activation while alpha-tocopherol did not increase the in vitro maturation rates, but improved the blastocyst rate. None of the three co-culture manner promoted the in vitro maturation and the cleavage of porcine DOs after parthenogenetic activation, but all the co-culture manners improved the blastocyst rates. Both Vitamin C and E enhance the in vitro developmental potential of porcine DOs. Co-culture increases the developmental potential of porcine DOs.

Effect of bone marrow mesenchymal stem cells on perforator skin flap survival area in rats.

Trans-territory perforator flaps are commonly used to reconstruct large defects of the soft tissues. The distal portion of the flap often becomes necrotic, however, as a result of the jeopardised vasculature of choke zone II. The trophic and vascular regenerative properties of bone marrow mesenchymal stem cells (BMSC) seemed to be a promising approach to prevent flaps becoming ischaemic. The purpose of our study is to evaluate the effects of BMSC on the survival of the three-territory perforator flap. The flap model was created based on the deep circumflex iliac vessel in rats. Eighteen rats were distributed, at random, into three groups. Immediately after the flaps were placed, groups were respectively given a single panniculus carnosus injection at choke zone II of either 1×105 (BMSCslow), 1×106 (BMSCshig) BMSC, or phosphate-buffered saline (PBS). On postoperative day seven, we assessed the gross view of the flap and survival. We also evaluated microvessels by histological examination and angiogenesis-related gene expression by quantitative real-time polymerase chain reaction. After high dosage of BMSC, the flap survival rate, diameter and density of microvessels, vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) levels were significantly higher in the BMSC treatment group than the control group. We therefore confirmed the positive effects of BMSC on the survival of multi-territory perforator flaps.