Eight isolates of Colletotrichum gloeosporioides isolated from host plants Cunninghamia lanceolata lanceolata and Euonymus japonichum, respectively, were cultured on MMC medium containing KCIO3 to select chlorate-resistant and nitrate non-utilizing mutants (Nit). All the Nit mutants obtained by this way belong to one of 3 kinds of the following phenotypes: the nitrate reductase structural locus (nit1), the nitrate-assimilation pathway-specific regulatory locus(nit3), and the molybdenum-containing cofactor locus(nitM). The higher mutation frequency on MMC amended with increasing concentration of KCIO3 was induced, and various nitrogen sources were able to influence production of the phenotypic classes. Seven of the 8 isolates tested were self-compatibility in which the two mutants with different phenotypes from the same isolate could genetically complement. The two isolates frail C. lanceolata belong to the same vegetative compatibility group (VCG), the other six isolates belong to distinctive VCGs.

MLFE (Micrococcus luteus fibrinolytic enzyme) is a fibrinolytic enzyme produced by Micrococcus luteus ML909 strain. The promoter and signal peptide-coding sequence of ?-amylase gene from Bacillus amyloliquefaciens DC-4 was cloned and fused to the sequence coding for mature peptide of MLFE (Gen-Bank: EU232121), forming the fusion gene called amymlfe. This hybrid gene was inserted into the Escherichia coli-Bacillus subtilis shuttle plasmid vector pSUGV4 and expression plasmid pSU-AmyMLFE was constructed. After transformation with B. subtilis WB600, transformant WB600/pSU-AmyMLFE was obtained and produced clear hydrolyzed zones on fibrin plates. The fibrinolytic activity in supernatants of WB600/pSU-AmyMLFE fermented for 24 hours was tested and found to be 238 UKU/mL. The results of SDS-PAGE analysis showed that there was indeed recombinant protein in supernatants. The Western blotting showed that the molecular weight of the expressed protein was the same as expected. These results indicate that the gene, amymlfe, is successfully expressed in B. subtilis WB600.

BACKGROUND: Jinguishenqi pills (JP) can ameliorate the symptoms of kidney deficiency, but the molecular mechanism of the effect is unclear.OBJECTIVE: To observe the differentially expressed genes of panic-induced kidney deficiency model mice treated with JP using suppression subtractive hybridization (SSH).DESIGN: Randomized control animal experiment.SETTING: Institute of Traditional Chinese Medicine Genetics, Chengdu University of Traditional Chinese Medicine; Sichuan Key Laboratory of Molecular Biology and Biotechnology, College of Life Science, Sichuan University.MATERIALS: This experiment was carried out at the Experimental Animal Center of Chengdu University of Traditional Chinese Medicine. Totally 30 BALB/c male adult mice were used in the experiment, and they were randomly divided into model group, JP-treated group and control group,with 10 in each.METHODS: Mice models of kidney deficiency were created in the mice of model group and JP-treated group with the methods of "terrifying mouse with cat" and "suspending mouse over water". Mice of the JP-treated group were intragastrically administrated of JP (produced in Tongrentang Pharmacy Co., Beijing, Z11020147, composed of Guizhi, Fuzi, Shudi, Shanyao,Shanzhuyu, Zexie, Fuling, Danpi, etc) 20-fold as human taken according to the bodyweight once per day. Modeling and administration were conducted in 14 days. Mice of normal control group were fed normally without any extra stimulation. Twenty-four hours after the last administration, 100 μL blood was collected respectively from mice of each group. Switch Mechanism at 5' end of RNA Template (SMART) technique was used in reverse transcription and amplification to obtain the whole length of total cDNA of mice in each group; Forward and reverse SSHs were used to acquire 6 groups of differential expression cDNA segments which were under 3 states(at kidney deficiency, after treatment and at normal physiological status).MAIN OUTCOME MEASURES: Effect of JP on gene expression of kidney deficiency mice RESULTS: ① Differentially expressed cDNA segments were found in each group; The differentially expressed cDNA profile of model mice had restored to that of the normal physiological state by JP treatment. cDNA of the same kind was used as the tester and driver of SSH, and almost all the homologous fragments were subtracted completely, which showed that the subtraction efficiency was very high. ② 6 cDNA libraries were acquired. CONCLUSION: Panic-induced kidney deficiency is related to differential expression of some genes. The gene expression of the kindey deficiency model mice could be affected by JP treatment,making it trend to normal physiological status.

Objective: To evaluate the effects of Xiaoer Feire Kechuan Granule for treating children with bronchial pneumonia. Methods: A randomly controlled trial was used. One hundred and twenty children with bronchial pneumonia were studied. The children in the experimental group were given Xiaoer Feire Kechuan Granule combined with basic treatment, and in the control group only treated by basic treatment, then the children with the disease in these two groups were observed to evaluate the general curative effect, traditional Chinese medicine (TCM) syndrome efficacy, symptoms of fever, and cough disappeared time before and after treatment of TCM primary symptom integra. Results: The overall efficacies in disease, syndromes curative effect, cough effect disappeared time, primary symptom score after therapy and hospital days for children in the two groups had statistical significance (P<0.05). Conclusion: Xiaoer Feire Kechuan Granule is applicable to the treatment of infantile pneumonia by antipyretic and antitussive function, promoting the pulmonary rales absorption, and shortening the course of treatment.

OBJECTIVETo evaluate the therapeutic effect of Parkinson's disease combined with overactive bladder syndrome (GAB) treated with combined therapy of oral administration of Tolterodine with low dose and electroacuponcture.

METHODSSixty cases of Parkinson's disease combined with GAB were randomly divided into a combined acupuncture and medication group (group A) and a medication group (group B), 30 cases in each group. In both groups, Madopar basic doses were same, and anticholinergic agents such as Artane were stopped. In group A, Tolterodine was orally taken for 1 mg, twice a day; Baihui (GV 20), Sishengcong (EX-HN 1) and Yintang (EX-HN 3) were punctured with electroacupuncture, once a day. In group B, Tolterodine was orally taken for 2 mg, twice a day. After 6 weeks, the changes of urination and UPDRS III scores were observed, and the adverse reactions were recorded in both groups.

RESULTSAfter treatment, the frequency of average urination of 24 hours, frequency of incontinence of 24 hours and average urine volume at a time were obviously improved (all P < 0. 01), of which, the above items in group A were superior to those in group B (all P < 0. 05) the UPDRSIII score in group A was superior to that in group B (P < 0.05). The adverse reactions in group A were less than those in group B.

CONCLUSIONThe therapeutic effect of Parkinson' s disease combined with GAB treated with combined therapy of Tolterodine with low dose and electroacupuncture is superior to that of complete dose of Tolterodine with oral administration, with less adverse reactions. And it also can improve the motor symptom of Parkinson's disease patients.

Adult ; Aged ; Benzhydryl Compounds ; therapeutic use ; Combined Modality Therapy ; Cresols ; therapeutic use ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; drug therapy ; therapy ; Phenylpropanolamine ; therapeutic use ; Tolterodine Tartrate ; Urinary Bladder, Overactive ; drug therapy ; therapy

Objective To investigate the correlation between neural tube defects (NTDs) and DACT1 gene, and provide the basic data for disease diagnosis and genetic counseling. Methods Blood samples were obtained from 163 NTDs patients and 480 unrelated healthy individuals. Mutation detection of DACT1 gene and DNA direct sequencing was carried out by PCR amplification. Bioinformatics analysis of these mutated loci was performed. Results Six mutations were found in NTDs patients, including 4 missense mutations (p.R45W, p.D142G, p.N356K and p.V702G). But these mutations were not found in 480 healthy individuals. Three mutated amino acid residues (p.45R, p.142D and p.356N) were highly conservative in evolution, and the mutated carriers were female patients, and suffered from anencephaly. Conclusion DACT1 gene mutation may be a risk factor of NTDs in Han population of northern China.

Objective: Based on observational, longitudinal and intervention study of common diseases among students in Jiangsu Province, this paper presents the current progress of two year follow up of myopia cohort regarding the association between growth parameters with progression of myopia among children and adolescents in areas with rapid economic growth. Methods: This survey adopted the stratified cluster sampling method for school selection. The full automatic computer optometry (TOPCON RM800) was used to track myopia related parameters for all participants from 2019 to 2020 under the condition of mydriasis (compound topicamide eye drops). Relationship between growth parameters of children and adolescents and the incidence and progression of myopia was analyzed by using Cox regression multiple statistical model. Results: The myopia rates of students from grade 1 to grade 3 in 2019 were 5.4%, 21.5% and 37.3% respectively. After one year, the myopia rates of all school stages increased to 25.3%, 43.3% and 58.1% respectively( χ 2=53.59, 49.63, 32.52, P <0.01). The mean diopter of right eye and left eye after mydriasis were ( 0.30± 1.24/0.39±1.26)D in 2019 and (-0.33±1.54/-0.19±1.55)D in 2020, respectively based on Cox multiple regression results, age ( HR =1.21, 95% CI =1.09-1.34), naked eye vision ( HR =0.08, 95% CI =0.07-0.11), height ( HR =0.98, 95% CI =0.97-0.99) showed a strong correlation with the incidence and progression of myopia( P <0.05). Conclusion Myopia is growing rapidly in the central region of Jiangsu Province. It is suggested that diopter, axial length, naked eye vision, age, height and other indicators should be included in the refractive archives of children and adolescents in the region.

Predicting protein-coding genes still remains a significant challenge. Although a variety of computational programs that use commonly machine learning methods have emerged, the accuracy of predictions remains a low level when implementing in large genomic sequences. Moreover, computational gene finding in newly sequenced genomes is especially a difficult task due to the absence of a training set of abundant validated genes. Here we present a new gene-finding program, SCGPred, to improve the accuracy of prediction by combining multiple sources of evidence. SCGPred can perform both supervised method in previously well-studied genomes and unsupervised one in novel genomes. By testing with datasets composed of large DNA sequences from human and a novel genome of Ustilago maydi, SCG-Pred gains a significant improvement in comparison to the popular ab initio gene predictors. We also demonstrate that SCGPred can significantly improve prediction in novel genomes by combining several foreign gene finders with similarity alignments, which is superior to other unsupervised methods. Therefore, SCG-Pred can serve as an alternative gene-finding tool for newly sequenced eukaryotic genomes. The program is freely available at http://bio.scu.edu.cn/SCGPred/.
Algorithms ; Chromosome Mapping ; methods ; Computational Biology ; methods ; Exons ; genetics ; Genes, Fungal ; genetics ; Genome, Fungal ; Genome, Human ; Humans ; Reproducibility of Results ; Software ; Ustilago ; genetics

Objective To investigate the methylation alteration of genomic DNA (gDNA) and its significance in pedigree neural tube defects (NTDs).Methods Twelve subjects from 3 NTDs pedigrees were enrolled in this study.NTDs patients were served as the case group,and their family members with normal phenotype were served as the control group.Peripheral vein blood was extracted,then gDNA was extracted.The extracted gDNA was treated with sodium bisulfite propagated as DNA segments in the way of whole genome amplification,which was put in I11umina Infinium human methylation 450k bead chip to perform hybridization,elution,extension,and imaging.The chip was scanned by iScan.Genome Studio was used to read the outcome.Illumina methylation analyzer software was used to analyze the methylation data.Results Gene differential methylation analysis showed that differential methylation sites only accounted for 0.2％ of the detected CpG sites and there were 617 differential hypermethylation sites (P ＜ 0.05),and 63 of them represented significant difference(P ＜ 1 × 10-4),including zinc finger E-box binding homebox 2,5,10-methylenetetrahydrofolate dehydrogenase 1 etc;there were 104 differential hypomethylation sites (P ＜ 0.05),and 65 of them represented significant difference (P ＜ 0.01),including Homeobox B7 and runt-related transcription factor 3 etc.Clustering analysis indicated that the tendency of DNA hypermethylation was consistent with NTDs patients,but the tendency of DNA hypomethylation was consistent with the controls.Conclusion In NTDs pedigree,the abnormal DNA methylation alterations may be the risk factor for NTDs occurrence.

ObjectiveThe development of new endovascular imaging techniques has optimized surgical treatment strategies. In this paper, we investigated the effect of rotational atherectomy (RA) guided by intravascular ultrasound (IVUS) on long-term postoperative follow-ups.MethodsClinical data of 211 patients who underwent RA in the Department of Cardiology, Nanjing Drum Tower Hospital from November 2011 to December 2018 were retrospectively analyzed, and they were divided into IVUS Group (116 patients) and Non-IVUS Group (95 patients) according to whether they underwent the guidance of intravascular ultrasound or not. Basic information of all patients, coronary artery lesions and details of operation and other clinical data were collected. The long-term prognosis of the patients was collected and compared by telephone or outpatient follow-ups.ResultsThe head diameter, average stent diameter and total hospitalization expenses of the IVUS Group were significantly higher than those of the Non-IVUS Group, and the differences were statistically significant [(1.53±0.19) mm vs (1.46±0.14) mm, P=0.001; (3.09±0.48) mm vs (2.87±0.30) mm, P<0.001; (57,300±18,300) yuan vs (49,300±16,600) yuan, P=0.001]. The all-cause case fatality rate and cardiogenic case fatality rate of the IVUS Group were lower than those of the Non-IVUS Group (7.5% vs 16.7%, P=0.048; 1.9% vs 11.9%, P=0.005); There were no significant differences in the incidence of long-term major adverse cardiovascular events (MACE), myocardial infarction, target vessel revascularization and non-fatal stroke between the two groups (P>0.05). Multivariate COX regression analysis showed that the cardiogenic mortality was significantly reduced in the IVUS Group (HR=0.10, 95%CI: 0.02~0.63, P=0.014), but there was no statistically significant difference between the two groups in the incidence of all-cause death and long-term MACE (P>0.05).ConclusionCompared with the Non-IVUS Group, IVUS-guided RA can significantly reduce the incidence of long-term cardiogenic death and total hospitalization expenses.