TCP family as plant specific transcription factor, plays an important role in different aspects of plant development. In order to screen TCP family members in tobacco, the homologous sequences of tobacco and Arabidopsis TCP family were identified by genome-wide homologous alignment. The physicochemical properties, phylogenetic relationships and cis-acting elements were analyzed by bioinformatics. The homologous genes of AtTCP3/AtTCP4 were screened, and RT-qPCR was used to detect the changes of gene expression upon 20% PEG6000 treatment. The results show that tobacco contains 63 TCP family members. Their amino acid sequence length ranged from 89 aa to 596 aa, and their protein hydropathicity grand average of hydropathicity (GRAVY) ranged from -1.147 to 0.125. The isoelectric point (pI) ranges from 4.42 to 9.94, the number of introns is 0 to 3, and the subcellular location is all located in the nucleus. The results of conserved domain and phylogenetic relationship analysis showed that the tobacco TCP family can be divided into PCF, CIN and CYC/TB1 subfamilies, and each subfamily has a stable sequence. The results of cis-acting elements in gene promoter region showed that TCP family genes contain low docile acting elements (LTR) and a variety of stress and metabolic regulation related elements (MYB, MYC). Analysis of gene expression patterns showed that AtTCP3/AtTCP4 homologous genes (NtTCP6, NtTCP28, NtTCP30, NtTCP33, NtTCP42, NtTCP57, NtTCP63) accounted for 20% PEG6000 treatment significantly up-regulated/down-regulated expression, and NtTCP30 and NtTCP57 genes were selected as candidate genes in response to drought. The results of this study analyzed the TCP family in the tobacco genome and provided candidate genes for the study of drought-resistance gene function and variety breeding in tobacco.
Nicotiana/genetics* ; Phylogeny ; Plant Breeding ; Amino Acid Sequence ; Arabidopsis ; Polyethylene Glycols

OBJECTIVE: To explore the genetic basis for a child with metachromatic leukodystrophy (MLD). METHODS: Clinical data of the patient was collected. Genomic DNA was extracted from peripheral blood samples of the child and his family members. Potential variant was screened by whole exome sequencing (WES), and candidate variant was verified by Sanger sequencing. The pathogenicity the variant was analyzed by multiple sequence alignment of the amino acid sequence and three-dimensional model prediction of its protein product. RESULTS: The child was found to harbor compound heterozygous variants c.257G>A (p.R86Q) and c.467del (p.G156Afs*6) of the ARSA gene, among which the c.467del (p.G156Afs*6) frameshift variation was unreported previously. Multiple sequence alignment showed that the site of the c.257G>A (p.R86Q) missense variant is highly conserved. Three-dimensional structure modeling analysis showed that the partial deletion due to the p.G156Afs*6 variant may cause significant alteration of the structure of ARSA protein. CONCLUSION The discovery of novel variant in ARSA has enriched the mutational spectrum of MLD and may facilitate the understanding of the genotype-phenotype correlation of MLD.
Cerebroside-Sulfatase/genetics* ; DNA ; Genetic Association Studies ; Humans ; Leukodystrophy, Metachromatic/genetics* ; Mutation

Objective:To analyze the correlation between recessive obesity and lifestyle in physical examination population.Methods:A total of 1 026 people with body mass index (BMI) of 18.5<-24.9 kg/m 2 who completed physical examination in the Health Management Center of Affiliated Hospital of Qingdao University from March 1, 2019 to September 31, 2019 were included in this study. Inbody770 body composition analyzer was used to check the percent body fat, and the subjects were divided into recessive obesity group (405 cases) and control group (621 cases) with the results. The results of routine physical examination were compared between the two groups. The data of five dimensions including marital status, education level, income, occupation and dietary habits were obtained with lifestyle questionnaire. The correlation between recessive obesity and lifestyle was analyzed by binary logistic regression. Results:The proportion of women, BMI, total cholesterol, low density lipoprotein cholesterol and waist-hip ratio were significantly higher in the recessive obesity group than those in the control group [65.2% vs 27.1%, (21.83±1.63) vs (21.56±1.74) kg/m2, 4.68 (4.18, 4.22) vs 4.39 (4.13, 4.83) mmol/L, 2.54 (2.08, 3.00) vs 2.24 (2.13, 2.78) mmol/L, 0.87 (0.84, 0.90) vs 0.82 (0.80, 0.86)]; while the systolic blood pressure, albumin and aspartate aminotransferase were significantly lower [(114.99±11.49) vs (118.97±11.84) mmHg (1 mmHg=0.133 kPa), (45.13±2.83) vs (46.37±2.60) g/L, 15 (12, 18) vs 16 (14, 20) U/L] (all P<0.05). Unmarried ( OR=0.200, 95% CI: 0.123-0.325), eating less white meat [occasionally eating, OR=0.565, 95% CI: 0.304-1.053; rarely eating, OR=0.186, 95% CI: 0.094-0.368], eating less spicy food (occasionally eating, OR=0.298, 95% CI: 0.171-0.519; rarely eating, OR=0.828, 95% CI: 0.487-1.408), drinking more water (1 000-2 000 ml/d, OR=0.366, 95%CI: 0.218-0.615; ≥2 000 ml/d, OR=0.176, 95% CI: 0.087-0.356) were negatively correlated with the occurrence of recessive obesity (all P<0.05). Eating takeout food frequently ( OR=4.639, 95% CI: 2.412-8.923), eating too much edible oil ( OR=10.900, 95% CI: 4.376-27.148), drinking beer ( OR=3.702, 95% CI: 2.290-5.982) and infrequent physical exercise (occasional, OR=13.417, 95% CI: 6.907-26.066; rarely, OR=28.290, 95% CI: 13.532-59.142) were positively correlated with the occurrence of recessive obesity (all P<0.05). Conclusions:There is a correlation between recessive obesity and the lifestyle in physical examination population. Attention should be paid to control the intake of white meat, spicy food, edible oil and beer, ensure the amount of drinking water, reduce the frequency of takeout food, and increase physical exercise to prevent recessive obesity.

Objective To investigate the features and characteristics of GLIS3 gene mutation in patients with congenital hypothyroidism(CH),and to establish the theoretical basis for gene diagnosis and prenatal diagnosis of CH.Methods Genomic DNA was extracted from peripheral blood leukocytes of 50 patients with CH who were collected from February 2007 to November 2016 in Shandong Province.The exon 2 to 11 of GLIS3 were amplified with 11 pairs of sequence specific primers designed by Primer 5.0.Polymerase chain reaction and the first generation of sequencing method(Sanger sequencing) were used to detect the mutation.Comparison of the sequencing results with the GLIS3 reference sequence (National Center for Biotechnology Information Reference Sequence:NC_000009.12) helped to screen gene mutations.Results The 50 CH patients included 22 boys and 28 girls,and the sex ratio was 1.0 ∶ 1.3.The mean age was (2.5 ± 0.5) years.Six cases (12％) had thyroid gland hypoplasia,23 cases (46％) had thyroid gland agenesis and 21 cases(42％) with ectopic thyroid gland.C2507A missense mutation was found in exon 10 of GLIS3 in a thyroid gland agenesis case,which might result in proline to glutamine substitution at codon 836.One mutant (rs780019691,c.C289T) was detected which was nonsense mutation (Arg→Stop) in another thyroid gland agenesis child.Conclusions The mutation rate of GLIS3 gene is very low in CH children of Shandong province.Further studies are needed to investigate the relationship between GLIS3 genotypes and clinical phenotypes.

Objective:To explore the inhibitory effect of dihydroartemisinin (DHA ) on the growth of neuroblastoma cells,and to clarify the anti-tumor mechanism of DHA.Methods:The experiment was divided into blank control group and DHA groups (the final concentrations of DHA were 0.05, 0.50, 5.00 and 50.00μmol·L-1 ).The proliferation rates of neuroblastoma SH-SY5Y cells after treated with DHA were examined by MTT assay;the changes of cell cycle of SH-SY5Y cells after treated with DHA were examined by flow cytometry;the expression levels of cyclin D1 and caspase-3 proteins were detected by ELISA and Western blotting methods.Results:The proliferation of SH-SY5Y cells 24,48,and 72 h after treated with different concentrations of DHA were inhibited.Compared with blank control group,the proliferation rates of SH-SY5Y cells in 0.50,5.00 and 50.00μmol·L-1 DHA groups were significantly decreased (P<0.05 or P<0.01).The density of cells was decreased with the increasing of DHA concentration.Compared with blank control group,the percentage of SH-SY5Y cells at SubG1 phase in 50.00μmol·L-1 DHA group was increased (P<0.05),and the percentage of cells at G0/G1 phase was increased first then was decreased;otherwise, the percentages of cells at S and G2/M phase were decreased.Compared with blank control group,the expression level of cyclin D1 protein in 50.00μmol·L-1 DHA group was decreased (P<0.05),but the expression level of caspase-3 protein in 50.00μmol· L-1 DHA group was increased (P<0.05).Conclusion:DHA could inhibit the proliferation through arresting the cell cycle and inducing the apoptosis of neuroblastoma cells.

Objective To screen the dual oxidase maturation factor 1 (DUOXA1) gene mutations in children with congenital hypothyroidism (CH) and thyroid goiter from Shandong Province,China,and to identify the gene mutation type and characteristics of DUOXA1 gene mutations in order to provide some evidence for gene diagnosis and therapy of CH.Methods A cohort of 52 cases of CH with thyroid goiter and 100 normal controls were selected according to neonatal screening system in Shandong Province whose genomic DNA was isolated from peripheral blood leukocytes with a standard phenol chloroform method.The whole coding sequence (CDS) of DUOXA1 gene was amplified with 8 pairs of sequence specific primers by using PCR.The PCR products were directly sequenced with Sanger sequencing to detect new mutations types of DUOXA1 gene.The sequencing data were compared to the DUOXA1 gene reference sequence(National Center for Biotechnology Information:RefSeq:NG_033105.1) to see if there was any mutation.Ax2 test was done for the gene frequency of discovered single nucleotide polymorphisms (SNP).Results There was no mutation in CDS of 52 CH patients with thyroid goiter and 100 normal controls.However,a SNP (rs75981505,c.398G ＞ T) which was an missense mutation and could lead to a change of the codon from CGC to CTC,was found in 9 CH patients with thyroid goiter and 11 normal controls in the exon 7.The corresponding amino acid arginine was replaced by histidine(p.Arg133His).There was no significant difference in the SNP rate between CH patients with thyroid goiter and normal controls (17.3％ vs 11.0％,x2 =1.24,P ＞ 0.05).Conclusion DUOXA1 gene mutation rate is very low which may not be the main cause of CH patients with thyroid goiter in the population of Shandong Province.

Objective To study the thyroid hormone receptor β(TRβ)gene mutation types and characteristics in children with congenital hypothyroidism(CH)and thyroid dysgenesis(TD)from Shandong Province,and to provide theoretical basis for gene diagnosis and prenatal diagnosis. Methods Sixty cases of TD patients of which genomic DNA were isolated from peripheral blood leukocytes were selected by neonatal screening system in Shandong Province. The exon 6 to 12 of TRβ gene were amplified with 8 pairs of sequence specific primers using PCR and the first generation of sequencing method(Sanger method)to detect mutation. The sequencing results were compared with the TRβ gene reference sequence[National Center for Biotechnology Information(NCBI)Reference Sequence:NC 000003. 12]to see whether there was a mutation. Results Analysis of TRβ in 60 cases of CH patients with TD revealed no mutation was demonstrated in exons 6 - 12,but 2 single nucleotide polymorphism(SNP)( rs 3752874,c. 735C ﹥ T;rs79220627, c. 162G ﹥ A)were detected. Through the analysis,the 2 SNP were all synonymous mutations(Phe→Phe;Ser→Ser), without the change of the amino acids. Conclusions TRβ mutation rate is very low,which may not be the main mutation type in CH patients with TD in Shandong Province.

Objective To investigate the associated factors of complete virological response (CVR) in Entecavir treatment among chronic hepatitis B (CHB) patients with positive HBeAg.Methods Data of 166 CHB patients with positive HBeAg receiving Entecavir treatment in Yantai Infectious Diseases Hospital during December 2009 and May 2014 were collected.Clinical data including age,gender,genotype,baseline alanine aminotransferase (ALT),HBV DNA load,HBsAg and HBeAg levels as well as CVR cumulative rates at different time points during Entecavir treatment were retrospectively analyzed.The cumulative rate of CVR was estimated using Kaplan-Meier method,and the difference in cumulative CVR rates was studied with Log-rank test.Cox's proportional hazards regression model was used to analyze the factors associated with CVR during Entecavir treatment.Results The cumulative rates of CVR during Entecavir treatment in HBeAg positive CHB patients were 54.5％ (87/157),74.3％ (106/129),80.2％ (109/119) and 86.8％ (110/112) at 48,96,144 and 192 weeks,respectively.Log-rank test showed that female patients and patients with genotype B,high baseline ALT level or low baseline HBV DNA load had higher CVR rates (x2 =15.601,11.542,17.021 and 10.094,all P ＜ 0.01).Cox's proportional hazards regression model showed that female was the only associated factor for CVR in Entecavir treatment among HBeAg positive CHB patients [hazard ratio (HR) =3.015,95％ confidence interval (CI):1.875-4.968,P ＜ 0.01].Conclusion CVR rate is increasing with the course of Entecavir treatment in HBeAg positive CHB patients,and CVR is more likely to occur in female patients.

Objective To investigate the status of life quality in the elderly in Shanxi Province Changzhi City, and to analyze its influencing factors. Methods The quality of life, activities of daily living and status of community services were measured simultaneously by 36-Item Short Form Health Survey (SF-36). Activities of Daily Living Scale, Community Healthy Needs Scale and survey results were used and analyzed using t-test, correlation and stepwise regression analysis. Results According to the criterion of life quality in the elderly which were proposed by Zhang Lei, the quality of life in the elderly scored 72. 1-117.0, reached 98.5%. The quality of life in the elderly was impacted by the ability of caring oneself, marital status, degree of culture, economic situation and so on. The demand rate for health guidance and periodic physical examination was higher. Conclusions The quality of life in the elderly is at a medium level. There is a wide gap between the demand of community healthy services and the utilization of community healthy services.

The study on the buds of Jasminum officinale L. var. grandiflorum was carried out to look for anti-HBV constituents. The isolation and purification were performed by HPLC and chromatography on silica gel, polyamide and Sephadex LH-20 column. The structures were elucidated on the basis of physicochemical properties and spectral analysis. Six iridoid glycosides were identified as jasgranoside B (1), 6-O-methy-catalpol (2), deacetyl asperulosidic acid (3), aucubin (4), 8-dehydroxy shanzhiside (5), and loganin (6). Jasgranoside B (1) is a new compound. Compounds 2-6 were isolated from Jasminum officinale L. var. grandiflorum for the first time.