The leucoanthocyantin reducase (LAR) gene, an important functional gene of catechins biosynthesis pathway, was cloned from Fagopyrum dibotrys (D.Don) Hara by degenerate PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA of FdLAR is 1 581 bp (GenBank accession: JN793953), containing a 1 176 bp ORF encoding a 391 amino acids protein, and its 3'-untranslated region has an obvious polyadenylation signal. The recombinant plasmid containing FdLAR completed ORF was transformed into E. coli BL21 (DE3). The target fusion peptide with molecular weight of 66 kD was expressed under the condition of 16 degrees C and induced by IPTG at final concentration of 1.0 mmol x L(-1). Bioinformation analysis indicated that the amino acid sequence of FdLAR showed great homology to other LAR with the NADB-Rossmann conversed domain in the N-terminus. Real-time quantitative PCR was used to detect the expression levels of FdLAR gene during different development periods. The determination of flavonoids contents in appropriate rhizomes showed that the relationship between FdLAR gene expression and the accumulation of flavonoids displayed different trends during vegetative growth and reproductive growth stages, suggesting that the FdLAR gene may be involved in the pathway of flavonoid metabolisms in Fagopyrum dibotrys.

Objective To observe the effect of Danshen in the treatment of severe hand-foot-and-mouth disease and investigate its mechanism.Methods One hundred and forty cases of children with severe hand foot and mouth diseases who hospitalized the intensive care unit were enrolled in the study from February 2012 to July 2014.The children were randomly divided into a control group and an observation group,with 70 cases in each group.The control group was given antiviral to reduce the intracranial pressure and intravenous immunoglobulin and the observation group was given Danshen in addition to the control group.The levels of neuron specific enolase (NSE)、S-100βprotein、TNF-α and IL-6 were detected at admission and checked again seven days after treatment.The changes of the above indicators and the clinical curative effect were compared before and after therapy.Results The total effective rate of the control group and the observation group was 80.0% and 92.9% respectively,showing statistical significance(P<0.05). After treatment, observation group of serum NSE (9.42 ± 2.56μg/Lvs. 11.71 ± 3.21μg/L,t=2.159), S-100β (177.34 ± 87.28 ng/Lvs. 286.14 ± 159.69 ng/L, t=2.315), TNF-α (15.98 ± 4.35 ng/Lvs. 23.17 ± 4.80 ng/L, t=4.297), IL-6 (41.72 ± 6.64 ng/Lvs. 52.05 ± 9.33 ng/L,t=3.492) , the level of were lower than those in the control group(P<0.05 orP<0.01). In the observation group, serum. The fever clearance time、the disappearance time of rash and the hospitalization time in the observation group (3.55 ± 1.02 d vs.4.55 ± 1.09 d, 7.14 ± 1.04 d) were shorter than those in the control group (4.46 ± 0.97 d vs.5.88 ± 1.44 d, 8.68 ± 1.06 d;t=5.409, 6.161, 8.677 respectively, P<0.01).Conclusions On the basis of conventional therapy,Danshen can effectively alleviate the systemic inflammatory response in children with severe hand foot and mouth diseases, reduce brain damage and improve the clinical efficacy.

Objective To assess mutations in the ALDH3A2 gene in two patients with Sj(o)gren-Larsson syndrome manifesting primarily as congenital ichthyosis,mental retardation and spastic paraplegia.Methods Two patients,a 2-year-old girl and a 1.5-year-old boy,with Sj(o)gren-Larsson syndrome were included in this study.None of their family members suffered from this disease.Peripheral blood samples were collected from the two patients,their family members (an elder brother and both parents),and 100 unrelated healthy controls.DNA was extracted from the blood samples,and subjected to PCR for the amplification of 10 encoding exons and their flanking sequences of the ALDH3A2 gene followed by DNA sequencing.Results A homozygous missense mutation c.325G ＞ A,which leads to the substitution of glycine by arginine at position 109,was detected in the ALDH3A2 gene of patient 1,whose parents and elder brother were heterozygous carriers of this mutation.The patient 2 carried compound heterozygous mutations,including c.1157A ＞ G (p.Asn386Ser) inherited from his father and c.1294A ＞ T (p.Arg432X) inherited from his mother.None of these mutations was detected in the unrelated healthy controls.Conclusion The homozygous mutation p.Gly109Arg and compound heterozygous mutations p.Asn386Ser and p.Arg432X present in these patients may be associated with clinical phenotypes of Sj(o)grenLarsson syndrome.

Objective To explore the tobacco control project based health education curriculum reform's effect of improving medical students' tobacco control.Method Choosing the students of two classes of students in preventive medicine specialty of Harbin Medical University as the research object, investigating the students respectively with the questionnaire prepared uniformly by 7 schools before and after the course.All data were coded by Epidata 3.0, and SPSS 18.0 was used to carry out the relevant information for the chi square test and t test.Result The sample size of baseline survey was114 and after the intervention was 87.The knowledge pass rate of the girls before and after the intervention were 87.3％ (48/55) and 98.2％ (54/55) respectively (P=0.000).The boys' awareness rate of the secondhand smoke safety level were 13.6％ (8/59) and 21.9％ (7/32) respectively, and the girls were 13.0％ (7/55) and 31.5％ (17/55) (P=0.021).Students interested in tobacco increased from 79.8％ (91/114) before intervention to 90.8％ (79/87) after intervention (P=0.033).Students willing to work in the tobacco related increased from 52.2％(59/114) before intervention to 66.7％ (58/87)after intervention (P=0.044).The girl who replied after intervention I am not smoking now, but won't smoke either when working in the disease control in the future reached as high as 96.4％ (53/55), but the students were lack of confidence to help people to stop smoking.Conclusion The reform of social practice activity curriculum designed for tobacco control project is effective, and it is urgent to strengthen the training of preventive medical students' smoking cessation skills and enhance their confidence.

Objective:To observe the changes of macular visual function after myopic foveoschisis (MF) and pars plana vitrectomy (PPV) combined with internal limiting membrane peeling and air filling.Methods:A single-center, retrospective study. From October 2018 to October 2019, 29 MF inpatients (32 eyes) in Tianjin Eye Hospital were included in this study. There were 3 males (4 eyes) and 26 females (28 eyes). The age was 63.00±3.45 years old. Equivalent spherical lens degree was -14.16±2.54 D, and axial length was 29.14±1.04 mm. Among them, 3 patients (3 eyes) had lamellar macular holes. All eyes underwent standard pars plana three-channel 25G PPV combined with internal limiting membrane peeling and air filling. Before surgery and at 1, 3, and 6 months after surgery, macular microperimetry was performed with a macular integrity assessment instrument, and the mean retinal sensitivitie (MS) within 10° of the macula, fovea 2° and 4° fixation rates (P1, P2), 63% and 95% bivariate contour ellipse area (BCEA) were recorded. The comparison of MS, P1, P2, 63%BCEA and 95%BCEA at different times before and after surgery was performed by paired t test; the comparison of fixation stability rate was performed by χ 2 test. Results:Compared with before surgery, there were significant differences in the improvement of MS in affected eyes at 1, 3 and 6 months after surgery ( t=-2.208, -3.435,-4.919; P=0.038, 0.002, 0.000). In the pairwise comparison at different times after surgery, only 6 months after surgery and 1 month after surgery were significantly different ( P=0.036). Compared with the preoperative P1, P2, 63%BCEA and 95%BCEA, the P1 and P2 of the eyes gradually increased after surgery, while the 63%BCEA and 95%BCEA gradually decreased, however, the difference was not statistically significant (P1: t=-1.595,-1.698,-1.966; P=0.125, 0.104, 0.062. P2: t=-1.622,-1.654,-1.707; P=0.119, 0.112, 0.102. 63%BCEA: t=1.410, 1.409, 1.553; P=0.172, 0.173, 0.135. 95%BCEA: t=1.412, 1.408, 1.564; P=0.172, 0.173, 0.132). Six months after surgery, all the eyes underwent anatomical repositioning of the macular area, and no serious complications such as full-thickness macular hole and macular hole retinal detachment were found. Conclusions:PPV with internal limiting membrane peeling and air filling is an effective and safe method for MF, and the macular function improved significantly within 6 months postoperatively.

Objective:To evaluate the role of 1, 4, 5-inositol triphosphate receptor (IP3R) in necroptosis of hippocampal neurons induced by sevoflurane anesthesia in aged rats.Methods:Sixty healthy male Sprague-Dawley rats, aged 18 months, weighing 500-600 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), sevoflurane anesthesia group (group S) and sevoflurane anesthesia + IP3R antagonist group (group S+ I). S and S+ I groups inhaled 2% sevoflurane for 5 h. In group S+ I, IP3 receptor antagonist 2-APB 3 mg/kg was intraperitoneally injected at 10 min before sevoflurane inhalation, and the equal volume of dimethyl sulfoxide was intraperitoneally injected in group C and group S. Morris water maze test was used to test the cognitive function on the day after the end of sevoflurane anesthesia.Then the animals were sacrificed and the brain tissues were obtained for microscopic examination of the pathological changes after HE staining and Nissl staining (with a light microscope) and for determination of the free calcium concentration ([Ca 2+ ] i) and rate of necroptosis of hippocampal neurons (by flow cytometry) and expression of IP3R, receptor-interacting protein kinase-1 (RIPK1), receptor-interacting protein kinase-3 (RIPK3) and phosphorylated mixed lineage kinase domain-like protein (p-MLKL) (by Western blot). Results:Compared with group C, the escape latency was significantly prolonged, the times of crossing the platform were reduced, the time of staying at the target quadrant was shortened, the [Ca 2+ ] i and necroptosis rate of hippocampal neurons were increased, and the expression of IP3R, RIPK1, RIPK3 and p-MLKL in hippocampal neurons was up-regulated in group S and group S+ I ( P<0.05). Compared with group S, the escape latency was significantly shortened, the times of crossing the platform were increased, the time of staying at the target quadrant was prolonged, the [Ca 2+ ] i and necroptosis rate of hippocampal neurons were decreased, and the expression of IP3R, RIPK1, RIPK3 and p-MLKL in hippocampal neurons was down-regulated in group S+ I ( P<0.05). Conclusions:The mechanism by which sevoflurane induces cognitive dysfunction may be related to the imbalance of calcium homeostasis caused by activation of IP3R and thus inducing programmed necrosis in aged rats.

Objective:To investigate predictive factors for successful endovascular recanalization in patients with non-acute symptomatic internal carotid artery occlusion (SICAO), to develop a decision tree model using the Classification and Regression Tree (CART) algorithm, and to evaluate the predictive performance of the model.Methods:Patients with non-acute SICAO received endovascular therapy at 8 comprehensive stroke centers in China were included retrospectively. They were randomly assigned to a training set and a validation set. In the training set, the least absolute shrinkage and selection operator (LASSO) algorithm was used to screen important variables, and a decision tree prediction model was constructed based on CART algorithm. The model was evaluated using the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow goodness-of-fit test and confusion matrix in the validation set.Results:A total of 511 patients with non-acute SICAO were included. They were randomly divided into a training set ( n=357) and a validation set ( n=154) in a 7:3 ratio. The successful recanalization rates after endovascular therapy were 58.8% and 58.4%, respectively. There was no statistically significant difference ( χ2=0.007, P=0.936). A CART decision tree model consisting of 5 variables, 5 layers and 9 classification rules was constructed using the six non-zero-coefficient variables selected by LASSO regression. The predictive factors for successful recanalization included fewer occluded segments, proximal tapered stump, ASITN/SIR collateral grading of 1-2, ischemic stroke, and a recent event to endovascular therapy time of 1-30 d. ROC analysis showed that the area under curve of the decision tree model in the training set was 0.810 (95% confidence interval 0.764-0.857), and the optimal cut-off value for predicting successful recanalization was 0.71. The area under curve in the validation set was 0.763 (95% confidence interval 0.687-0.839). The accuracy was 70.1%, precision was 81.4%, sensitivity was 63.3%, and specificity was 79.7%. The Hosmer-Lemeshow test in both groups showed P>0.05. Conclusion:Based on the type of ischemic event, the time from the latest event to endovascular therapy, proximal stump morphology, the number of occluded segments, and the ASITN/SIR collateral grading constructed the decision tree model can effectively predict successful recanalization after non-acute SICAO endovascular therapy.

Objective:To investigate the predictive value of fluid-attenuated inversion recovery (FLAIR) signal strength ratio (SIR) in onset time≤4.5 h in patients with acute ischemic stroke.Methods:A retrospective analysis was performed; 180 acute ischemic stroke patients admitted to Department of Neurology, Nanjing Hospital Affiliated to Nanjing Medical University from January 2020 to June 2023 were chosen. Hypoperfusion intensity ratio (HIR) was used to evaluate the collateral circulation (poor collateral circulation: HIR≤0.4; good collateral circulation: HIR>0.4); clinical data and imaging indexes between poor collateral circulation and good collateral circulation groups were compared. Univariate and multivariate Logistic regressions were used to analyze the influencing factors for onset time≤4.5 h in patients with acute ischemic stroke. Correlation between SIR and onset time was analyzed in patients with acute ischemic stroke. Role of HIR as agency between SIR and onset time was explored. Receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of SIR and diffusion weighted imaging (DWI)-FLAIR mismatch in onset time≤4.5 h in acute ischemic stroke patients.Results:Of the 180 patients, 100 were into the good collateral circulation group and 80 were into the poor collateral circulation group; compared with the good collateral circulation group, the poor collateral circulation group had significantly higher percentage of patients with hyperlipidemia, larger DWI infarction volume before treatment, larger perfusion weighted imaging (PWI)-DWI mismatch volume and higher SIR ( P<0.05). In these 180 patients, 76 had onset time≤4.5 h and 104 had onset time>4.5 h. Univariate Logistic regression analysis showed that hyperlipidemia, DWI infarct volume before treatment, DWI-FLAIR mismatch, HIR and SIR were influencing factors for onset time≤4.5 h in acute ischemic stroke patients ( P<0.05). Multivariate Logistic regression analysis showed that hyperlipidemia ( OR=6.654, 95% CI: 5.751-8.824, P<0.001), HIR ( OR=0.724, 95% CI: 0.521-1.321, P=0.041) and SIR ( OR=739.881, 95% CI: 383.296-14 258.065, P<0.001) were independent influencing factors for onset time≤4.5 h in acute ischemic stroke patients. Pearson correlation analysis showed that SIR was positively correlated to onset time in patients with acute ischemic stroke ( r=0.420, P<0.05), and SIR was positively correlated to onset time in patients from poor collateral circulation group ( r=0.781, P<0.05). ROC curve showed that AUC of SIR in predicting onset time≤4.5 h was 0.917 (95% CI: 0.814-1.000, P<0.001) and that of DWI-FLAIR mismatch in predicting onset time≤4.5 h was 0.530 (95% CI: 0.509-0.757, P=0.075) in poor collateral circulation group, enjoying significant difference in predictive efficacy. Conclusion:Acute ischemic stroke patients with low HIR and SIR have higher odds of onset time≤4.5 h; SIR can more accurately predict the onset time in these patients with poor collateral circulation.