OBJECTIVE:To establish the gas chromatography (GC) fingerprint for the petroleum ether part of Areca peel and provide reference for the quality evaluation. METHODS:GC was conducted to establish the fingerprint for the petroleum ether part of Areca peel from 10 different areas,the cluster analysis and similarity of fingerprint data were conducted to study the similarity of GC fingerprint of Areca peel. RESULTS:There were totally 8 common peaks. All the medicine materials had the characteristics,but there were differences among the relative peak area. CONCLUSIONS:The GC fingerprint has high importance and precision and can be used for the quality control of Areca peel.

To construct CRISPR/Cas9 vectors for the editing of SmPAL1 in the phenylpropane metabolic pathway of Salvia miltiorrhiza, CIRSPR/Cas9 target sites of SmPAL1 were designed by online software. Its target efficiencies were detected in vitro by enzyme digestion and sequences with highly efficiency were constructed into CRISPR/Cas9 vectors. Three possible CRISPR target sequences (SmPAL1-g1, SmPAL1-g2, SmPAL1-g3) were designed and the enzyme digestion efficiencies were 53.3%, 76.6% and 10.0%. SmPAL1-g1 and SmPAL1-g2 were constructed into vector VK005-03 named as VK005-03-g1 and VK005-03-g2. The results of sequencing showed that the two CRISPR/Cas target sequences were all constructed into VK005-03. Here we first laid the foundation for the study of SmPAL1 and provided an effective strategy for the screening of sgRNA.
CRISPR-Cas Systems ; Clustered Regularly Interspaced Short Palindromic Repeats ; Metabolic Networks and Pathways ; genetics ; Salvia miltiorrhiza ; chemistry ; genetics

ObjectiveTo assess the relationship between alcoholic fatty liver disease (AFLD) and carotid intima-media thickness (CIMT) and the influencing factors for CIMT. MethodsA cross-sectional study was conducted in 147 males who were inpatients or had physical examination at the Taishan Hospital in Shandong Province during January 2012 to December 2013. The participants, including 136 drinking cases and 11 non-drinking cases, were divided into mild AFLD group (A, 45 cases), moderate to severe AFLD group (B, 58 cases), and non-AFLD group (C, 44 cases). Comparison of mean values between groups was performed using one-way ANOVA, and pairwise comparisons were performed using the LSD test (for homogeneity of variance) or Tamhane's test (for heterogeneity of variance). Results(1) CIMT was significantly higher in group B than in group C (P=0.001). (2) BMI was significantly higher in group A than in group C (P=0.029). Total cholesterol, triglyceride, very-low-density lipoprotein, apolipoprotein B, and uric acid levels significantly rose in groups A and B compared with group C (P＜0.05). (3) Gamma-glutamyl transpeptidase (GGT) level was significantly higher in group A than in group C (P=0001), whereas alanine transaminase, aspartate aminotransferase, and GGT levels were significantly higher in group B than in group C (P=0.023, 0.003, and 0.000, respectively). (4) Serum creatinine level was significantly lower in groups A and B than in group C (P=0007 and 0.005, respectively). ConclusionAFLD can cause the increase in CIMT, resulting in metabolic syndrome-like changes and liver/kidney dysfunction. With increasing severity of AFLD, CIMT increase becomes more significant. Thus, AFLD can be used as an indicator for predicting CIMT increase in human population.

Objective To investigate the feasibility and efficacy of carotid endarterectomy and carotid artery stenting in the treatment of symptomatic severe carotid stenosis. Methods Retrospectively analyzed 203 patients with symptomatic severe carotid artery stenosis who received CEA or CAS treatment in Taishan Affiliated Hospital of Shandong First Medical University hospital from January 2016 to December 2018,The postoperative complications,and 1 year follow-up outcomes were recorded. Results A total of 203 patients with symptomatic severe carotid stenosis were included,and they were divided into the CAS group (n=132) and the CEA group (n=71). There were no statistically significant differences in age,gender,risk factors,blood pressure,etc. (P>0.05). In the analysis of perioperative complications,there were 3 cases of neck hematoma in CEA group (P=0.017). The proportion of stroke within 30 days in the CAS group was significantly higher than that in the CEA group (P=0.034). Results Of 1-year follow-up showed that the restenosis rate of CAS group was significantly higher than that of CEA group (P=0.047). There was no significant difference between the two groups in terms of myocardial infarction,death and stroke. Conclusion Both CAS group and CEA group can safely and effectively treat symptomatic severe carotid artery stenosis. In terms of perioperative complications,neck swelling in the CEA group was significantly higher than CAS group,and the proportion of stroke within 30 days and restenosis within 1-year flow up in the CAS group was significantly higher than that in the CEA group.

To study the correlation between 50% tissue-culture infective dose (TCID50) value and p27 antigen S/P value of Avian leukosis virus subgroup J and discuss their significance, chicken embryo fibroblast (CEF) cells were inoculated with Avian leukosis virus subgroup J strain NX0101 and samples were tested continuously for ten days after changing maintenance media. The correlation between TCID50 and p27 antigen S/P value of ten days were then analysized. Simultaneously, DF-1 cells were inoculated with NX0101 and passaged to 20 generations. Samples taken from 1st generation, 5th generation, 10th generation, 15th generation and 20th generation were tested for the TCID50 titer and the p27 antigen S/P value separately. A significant Pearson correlation was found between them in CEF cells (r = 0.85277; P < 0.0001) and in DF-1 cells (r = 0.93000; P = 0.0220). This study provided an important parameter for predicting TCID50 by detecting the p27 antigen S/P value.
Animals ; Avian Leukosis ; virology ; Avian Leukosis Virus ; immunology ; pathogenicity ; Chick Embryo ; Fibroblasts ; virology ; Proliferating Cell Nuclear Antigen ; analysis ; immunology ; Viral Load ; immunology

Depression is a mental or affective disorder， whose occurrence is closely related to the physical， psychological， and social environmental factors. Clinically， patients with depression often present with significant and long-lasting blue mood， impaired thinking and cognitive functions， reduced mobility， and even self-harm and suicide attempts. The pathogenesis of depression is complex and diverse， and there mainly exist the neurotransmitter hypothesis， immunodeficiency hypothesis， hypothalamus-pituitary-adrenal （HPA） axis activation hypothesis， brain-derived neurotrophic factor （BDNF） hypothesis， and intestinal flora hypothesis. In western medicine， it is mostly treated with antidepressant drugs， but the resulting side effects and repeated attack cannot be ignored. Traditional Chinese medicine（TCM） has obvious advantages in dealing with depression. The advocates of treating "Qi"， "phlegm"， "stasis"， and "five zang organs" have all been proved effective. Depression is characterized by a long course of disease and repeated attack， which coincides with the concept of "deficiency" in TCM. Tonifying deficiency is a method for treating depression based on TCM syndrome differentiation. Under the guidance of this principle， the formulated prescriptions will produce the desired effects. This article reviewed the related clinical and experimental studies on depression treatment via deficiency tonification in recent years， and summarized the corresponding prescriptions for benefiting Qi， replenishing blood， nourishing Yin， and tonifying yang， respectively， so as to enrich the TCM theory of depression and provide new ideas for its clinical treatment.

Edible bird's nest contains lots of glycoproteins. The glycosylation inhomogeneity for glycoprotein often results in wide range of molecular weight and the difficulty for protein separation and charaterization. In this paper, proteins in the edible bird's nest were extracted using multiple extractions, and then digested by PNgase F and trypsin. The digest mixture was separated with HPLC, and peptides were identified based on MS/MS data searching. The results indicated that the extracted proteins were amount to 79.7% of total protein in the edible bird's nest. More than 20 species of peptides in the digested mixture were identified. The sequences of these peptides showed similarity with some proteins from Swiss-prot. The research indicated that deglycosylation, tryptic digestion coupled with HPLC-MS/MS is a proper strategy for characterization of proteins in the edible bird's nest.
Amino Acid Sequence ; Animals ; Birds ; Chromatography, High Pressure Liquid ; Glycoproteins ; chemistry ; Mass Spectrometry ; Medicine, Chinese Traditional ; Peptide Fragments ; chemistry ; isolation & purification ; metabolism ; Proteolysis

Mood disorders/psychosis have been associated with dysfunctions in the default mode network (DMN). However, the relative contributions of DMN regions to state and trait disturbances in pediatric bipolar disorder (PBD) remain unclear. The aim of this study was to investigate the possible mechanisms of PBD through brain imaging and explore the influence of psychotic symptoms on functional alterations in PBD patients. Twenty-nine psychotic and 26 non-psychotic PBD patients, as well as 19 age- and sex-matched healthy controls underwent a resting-state functional MRI scan and the data were analyzed by independent component analysis. The DMN component from the fMRI data was extracted for each participant. Spearman's rank correlation analysis was performed between aberrant connectivity and clinical measurements. The results demonstrated that psychotic PBD was characterized by aberrant DMN connectivity in the anterior cingulate cortex/medial prefrontal cortex, bilateral caudate nucleus, bilateral angular gyri, and left middle temporal gyrus, while non-psychotic PBD was not, suggesting further impairment with the development of psychosis. In summary, we demonstrated unique impairment in DMN functional connectivity in the psychotic PBD group. These specific neuroanatomical abnormalities may shed light on the underlying pathophysiology and presentation of PBD.

BACKGROUND: MicroRNA-20a (miR-20a) is dysregulated in many types of malignancies, including human hepatocellular carcinoma (HCC), but its expression level and functional significance in HCC are still disputed. We aimed to study the role of miR-20a-5p in HCC and its downstream molecular mechanisms. METHODS: We used real-time polymerase chain reaction to detect the expression of miR-20a-5p and runt-related transcription factor 3 ( RUNX3 ) in HCC and paraneoplastic tissue, transfected Huh7 and highly metastatic human hepatocellular carcinoma (MHCC97H) cells. A live cell workstation was used to observe the proliferation and migration of transfected cells. The invasiveness of transfected cells was verified by Transwell assay. Cell apoptosis was detected by flow cytometry. The expression levels of proteins after transfection were measured using simple western immunoblot measurements. Gene expression profiles between HCC and normal samples were obtained from The Cancer Genome Atlas. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment results were processed by the database for annotation, visualization and integrated discovery. Potential target genes of miR-20a-5p were predicted to further investigate how miR-20a-5p regulates epithelial-mesenchymal transition (EMT) in HCC. RESULTS: MiR-20a-5p was significantly highly expressed in HCC tissues, and overexpression of miR-20a-5p significantly promoted HCC cell proliferation, migration, and invasion and inhibited apoptosis in vitro. The protein expression of E-cadherin was decreased and that of vimentin was increased after overexpression of miR-20a-5p in HCC cells. We discovered the intersection of genes from miRDB, miR TarBase, and TargetScan, obtained 397 target genes and finally focused on RUNX3. RUNX3 was not only reduced in HCC specimens but also drastically reduced in HCC cells overexpressing miR-20a-5p. RUNX3 expression decreased with elevated miR-20a-5p, which activated downstream EMT signaling and promoted cell proliferation, migration, and invasion. CONCLUSIONS Since RUNX3 is involved in EMT in HCC, as proven by previous research, our findings provide further evidence for a novel regulatory pathway comprising the miR-20a/RUNX3/EMT axis that upregulates EMT signaling and enhances the migration of HCC cells.
Humans ; Carcinoma, Hepatocellular/pathology* ; Epithelial-Mesenchymal Transition/genetics* ; Liver Neoplasms/pathology* ; Gene Expression Regulation, Neoplastic/genetics* ; MicroRNAs/metabolism* ; Cell Movement/genetics* ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Core Binding Factor Alpha 3 Subunit/metabolism*

OBJECTIVETo summarize the treatment status of gastric gastrointestinal stromal tumor (GIST) in Shandong province,by analyzing the clinicopathological features and prognostic factors.

METHODSClinicopathological and follow-up data of 1 165 patients with gastric GIST between January 2000 and December 2013 from 23 tertiary referral hospitals in Shandong Province were collected to establish a database. The risk stratification of all cases was performed according to the National Institutes of Health(NIH) criteria proposed in 2008. Kaplan-Meier method was used to calculate the survival rate. Log-rank test and Cox regression model were used for univariate and multivariate prognostic analyses.

RESULTSAmong 1 165 cases of gastric GIST, 557 were male and 608 were female. The median age of onset was 60 (range 15-89) years. Primary tumors were located in the gastric fundus and cardia in 623 cases(53.5%), gastric body in 346 cases(29.7%), gastric antrum in 196 cases(16.8%). All the cases underwent resection of tumors, including endoscopic resection (n=106), local resection (n=589), subtotal gastrectomy(n=399), and total gastrectomy(n=72). Based on the NIH risk stratification, there were 256 cases (22.0%) at very low risk, 435 (37.3%) at low risk, 251 cases (21.5%) at intermediate risk, and 223 cases (19.1%) at high risk. A total of 1 116 cases(95.8%) were followed up and the median follow-up period was 40 (range, 1-60) months. During the period, 337 patients relapsed and the median time to recurrence was 34 (range 1-60) months. The 1-, 3-, and 5-year survival rates were 98.6%, 86.1% and 73.4%, respectively. The 5-year survival rates of patients at very low, low, intermediate, and high risk were 93.1%, 85.8%, 63.0% and 42.3% respectively, with a statistically significant difference (P=0.000). Multivariate analysis showed that primary tumor site (RR=0.580, 95%CI:0.402-0.835), tumor size (RR=0.450, 95%CI:0.266-0.760), intraoperative tumor rupture(RR=0.557, 95%CI:0.336-0.924), risk classification (RR=0.309, 95%CI:0.164-0.580) and the use of imatinib after surgery (RR=1.993, 95%CI:1.350-2.922) were independent prognostic factors.

CONCLUSIONSThe choice of surgical procedure for gastric GIST patients should be based on tumor size. All the routine procedures including endoscopic resection, local excision, subtotal gastrectomy and total gastrectomy can obtain satisfactory curative outcomes. NIH classification has a high value for the prediction of prognosis. Primary tumor site, tumor size, intraoperative tumor rupture, risk stratification and postoperative use of imatinib are independent prognostic factors in gastric GIST patients.