1-Deoxy-D-xylulose-5-phosphate synthase (DXS), the first key enzyme in 2-methyl-D-erythritol-4-phosphate (MEP) pathway, catalyzes the condensation of glyceraldehyde-3-phosphate with pyruvate to 1-deoxy-xylose-5-phosphate (DXP). In this study, PgDXS1, PgDXS2, and PgDXS3 genes were cloned from the root of Platycodon grandiflorum (P. grandiflorum). The open reading frame (ORF) of PgDXS1, PgDXS2, and PgDXS3 were 2 160, 2 208, and 2 151 bp in full length, encoding 719, 735, and 716 amino acids, respectively. Homologous alignment results showed a high identity of PgDXSs with DXS in Hevea brasiliensis, Datura stramonium and Stevia rebaudiana. The recombinant expression plasmids of pET-28a-PgDXSs were constructed and transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and the induced proteins were successfully expressed. Subcellular localization results showed that PgDXS1 and PgDXS2 were mainly located in chloroplasts, and PgDXS3 was located in chloroplasts, nucleus and cytoplasm. The expression of three DXS genes in different tissues of two producing areas of P. grandiflorum were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves of P. grandiflorum from Taihe. Under methyl jasmonate (MeJA) treatment, the expression levels of three PgDXS genes showed a trend of first decreasing and then increasing at different time points (3 - 48 h), and the activity of DXS showed a trend of first increasing and then decreasing in three tissues of P. grandiflorum. This study provides a reference for further elucidating the biological function of PgDXS in terpenoid synthesis pathway in P. grandiflorum.

The "student-centered" flipped classroom teaching model can improve students' academic achievement, improve cognition, and cultivate innovation ability. However, it has obvious deficiencies in the integrity and systematization of knowledge as well as education. The traditional teaching concept based on " teacher-centered" has the unique advantages of systematization of knowledge learning and education. Therefore, we integrated the merits of these two different teaching models and introduced the semi-flipped classroom teaching model into the Biochemistry teaching of 2020 stomatology, pharmacy and preventive medicine majors in Cheeloo Medical College, Shandong University, compared with the traditional teaching of 2019 same majors. The data on the improvement of students ' academic achievement and self-cognition were analyzed. The results showed that the students' achievements of the semi-flipped classroom teaching model were better than those of the traditional teaching (P<0. 01). The students' self-cognitions were significantly improved after the implementation of semi-flipped classroom teaching (P<0. 01 or P<0. 05). This study provides a reference for the related teaching and research work in medical colleges.

Classical famous prescription Dihuang Yinzi is widely used in modern clinical practice，and can treat many kinds of diseases，especially the diseases of nervous system in internal medicine. Its clinical effect is accurate，but it has not been converted into Chinese patent medicine preparations. Therefore，the authors have collected ancient traditional Chinese medicine（TCM） literatures of Dihuang Yinzi by the methods of bibliometrics，and selected and sorted out 254 pieces of effective data， involving 144 ancient books of TCM，and systematically summarized and analyzed the historical development origin，main treatment syndrome，formula making principle，dosage，preparation method，decoction method and medicine taking method of Dihuang Yinzi，in order to provide the ancient literary evidence support for the development and clinical application of classic famous prescriptions. It is found that Dihuang Yinzi was from Xuanming Lunfang written by LIU He-jian，a doctor of Jin dynasty. It was composed of 12 kinds of herbs，namely Rehmanniae Radix Praeparata，Morindae Officinalis Radix，Corni Fructus，Cistanches Herba，Dendrobii Caulis，Aconm Lateralis Radix Praeparaia，Schisandrae Chinensis Fructus，Cinnamomi Ramulus，Poria，maimendong，Acori Calami Rhizoma and Polygalae Radix, and mainly used for the treatment of Yinfei. The later records of Dihuang Yinzi mostly followed the prescription composition and main treatment set forth in Xuanming Lunfang，and its clinical application was expanded. In the 199 articles with the indications for disease treatment，Yinfei was the most commonest indication, and took up about half of the total，which was followed by stroke，taking up about two fifths of the total. It was also used for the treatment of sudden aphonia，flaccidity syndrome，vertigo，enuresis. Dihuang Yinzi has a wide range of treatment，but the pathogenes is always belongs to "the deficiency of water and fire in the kidney". The recipe of Dihuang Yinzi was unique，and can be used to treat both the upper and lower parts of the body，as well as both the outward symptoms and root causes of an illness at the same time， in particular，it mainly focuses on the treatment of the lower and the root. Among the 56 literatures with drug dosage records，about one third of them inherited the records of Xuanming Lunfang: "Equal division，the top is the end，3 qian for each dose." The dosage was generally light. The preparations are mostly decoction and boiled powder. In the decocting and taking methods，it was suggested that "turbid medicine shall be boiled for a short time，and taken after several boilings，with no limit to time."

Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI：3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.

OBJECTIVETo evaluate the efficacy and safety of endoscopic mucous resection with transparent cap (EMR-Cap) and endoscopic multi-band mucosectomy (MBM) in the treatment of early esophageal cancer and precancerous lesion.

METHODSA retrospective study was performed to review 30 EMR-Cap cases from December 2008 to December 2009 and 32 MBM cases from January 2010 to January 2011 of early esophageal cancer and precancerous lesions. The differences between these two techniques in efficacy, safety, and cost were compared.

RESULTSIn EMR-Cap group, the median resection time was 26(10-56) min and median procedure time was 43(22-81) min, significantly longer than those in MBM group [10(7-18) min and 32(28-45) min, P=0.036 and 0.038, respectively]. There were no significant differences between the two groups in total thickness and depth of resected lesions (P>0.05). In EMR-Cap group, the median cost was significantly higher than that of MBM group [(5466±354) vs. (4014±368) RMB, P=0.008)].

CONCLUSIONSEMR-Cap and MBM are minimally invasive, safe and effective methods in the treatment of early esophageal cancer and precancerous lesions. Compared to the EMR-Cap, MBM is simple with shorter treatment time and lower cost.

Aged ; Endoscopy ; methods ; Esophageal Neoplasms ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mucous Membrane ; surgery ; Precancerous Conditions ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo study the effect of substrate stiffness on proliferation, migration of fibroblast and integrin β(1) expression in fibroblast.

METHODSFibroblasts were inoculated on silicon substrate with stiffness of (16.2 ± 0.5), (19.8 ± 1.1), and (200.1 ± 2.6) kPa. After being cultured for 5 days or 6 days, cells were counted and cell proliferative activities (recorded as absorbance value) were assessed with methyl thiazolyl blue (MTT). After being cultured for 3 days, cell cycle was detected and proliferation index (PI) was calculated. The cell scratch test was used for determination of cell migration rate on post scratch day (PSD) 0 (the day of scratch), 1, 2, and 3. After being cultured for 2 days, the expression of integrin β(1) was determined by flow cytometry with fluorescence. Data were processed with one-way analysis of variance.

RESULTS(1) The proliferative speed and proliferative activity of fibroblasts were all increased along with the increase in substrate stiffness. PI of fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5), (19.8 ± 1.1), and (200.1 ± 2.6) kPa was respectively 24.8%, 27.4%, 32.4%. On PSD 2, migration rate of fibroblasts inoculated on silicon substrate with stiffness of (19.8 ± 1.1) and (200.1 ± 2.6) kPa was respectively (91.4 ± 5.1)%, (100.0 ± 1.3)%, which were higher than that of fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5) kPa [(55.8 ± 6.8)%, with F value respectively 3.5, 4.0, P values all below 0.01]. (3) The expression rate of integrin β(1) in fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5) kPa was the lowest (43.22%), and that in fibroblast inoculated on silicon substrate with stiffness of (200.1 ± 2.6) kPa was the highest (81.26%).

CONCLUSIONSSubstrate stiffness may have a great effect on proliferation and migration of fibroblast during the process of wound healing and scar formation, which can be related to regulation of integrin β(1) expression.

Cell Movement ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; pathology ; Humans ; Integrin beta1 ; metabolism ; Mechanical Phenomena ; Silicon

OBJECTIVETo observe the clinical effectiveness of a topical application of Xiaozheng Zhitong: Paste (, XZP) in alleviating the cancerous pain of patients with middle/late stage cancer

METHODSBy: adopting a random number table, 124 patients enrolled were randomized into the treatment group (64 patients) and the control group (60 patients). In addition to the basic therapy [including the three-ladder (3L) analgesia] used in both groups, topical application of XZP was given to patients in the treatment group for pain alleviation. The analgesic efficacy was recorded in terms of pain intensity, analgesia initiating time and sustaining time, and the optimal analgesic effect revealing time. Meanwhile, the quality of life (QOL) and adverse reactions that occurred in patients were recorded as well.

RESULTSThe total effective rate in the treatment group was: 84.38% (54/64), and in the control group it was 88.33% (53/60), showing no significant difference between them (P>0.05), but the analgesia initiating time and the optimal analgesia effect revealing time in the treatment group were significantly shorter (both P<0.01). Moreover, XZP was better in improving patients' QOL, showing more significant improvements in the treatment group than those in the control group in aspects of mental condition, walking capacity, working capacity, social acceptability, sleep and joy of living (P<0.05 or P<0.01). Lower incidence of adverse reactions, such as nausea, vomiting, mouth dryness, dizziness, etc., especially constipation, was noted in the treatment group (P<0.05 or P<0.01).

CONCLUSIONApplying an external compress: of XZP showed a synergistic action with 3L analgesia for shortening the initiating time and the optimal effect revealing time, and could evidently enhance patients' QOL with fewer adverse reactions.

Aged ; Analgesics ; adverse effects ; therapeutic use ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Emollients ; adverse effects ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Neoplasms ; complications ; drug therapy ; Pain ; complications ; drug therapy ; Phytotherapy ; Quality of Life ; Time Factors ; Treatment Outcome

Coronary Restenosis ; pathology ; Fibroblasts ; metabolism ; pathology ; Fibrosis

OBJECTIVETo evaluate the value of transbronchial needle aspiration (TBNA) combined with transesophageal endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) in the diagnosis of mediastinal and pulmonary hilar lesions as well as in the lymph node staging (N staging) of lung cancer.

METHODS129 patients with mediastinal and pulmonary hilar lesions underwent either TBNA or EUS-FNA with cytological needle aspiration. The samples obtained from TBNA or EUS-FNA were examined by both cytologiy and histopathology.

RESULTSOf the 129 patients, 59 underwent TBNA and 70 EUS-FNA. The diagnostic rate were 84.7% (50/59) by TBNA and 94.3% (66/70) by EUS-FNA, resepectively. The diagnosis of 116 (89.9%) patients were confirmed by either TBNA or EUS-FNA. The pathological and cytological diagnostic rates were 92.2% (107/116) and 88.0% (102/116), resepectively. The diagnostic rate was elevated by 8.4% (9/107) through pathological examination. The histological classification rates by cytological and pathological examination were 73.8% (76/116) and 89.3% (92/103), respectively. The diagnostic rate of histological classification was elevated by 35.5% (27/76) through pathological examination.

CONCLUSIONThe combination of TBNA and EUS-FNA can improve the diagnostic rate for wider mediastinal and pulmlonary hilar lesions. Pathological examination of the samples obtained from the TBNA and EUS-FNA can elevate not only the rate of diagnosis but also the rate of histological classification.

Adenocarcinoma ; diagnostic imaging ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; methods ; Biopsy, Needle ; Carcinoma, Squamous Cell ; diagnostic imaging ; pathology ; Endosonography ; methods ; Female ; Humans ; Lung Neoplasms ; diagnostic imaging ; pathology ; Lymph Nodes ; diagnostic imaging ; pathology ; Lymphatic Metastasis ; Male ; Mediastinal Neoplasms ; diagnostic imaging ; pathology ; secondary ; Mediastinum ; Middle Aged ; Neoplasm Staging ; Small Cell Lung Carcinoma ; diagnostic imaging ; pathology ; Young Adult

OBJECTIVETo study the expression levels of monokine induced by interferon-gama; (Mig) mRNA and its association with HBV DNA and alanine aminotransferase (ALT) in patients with chronic hepatitis B.

METHODSThe level of Mig mRNA in peripheral blood mononuclear cells (PBMCs) was dynamically detected with real-time quantitative PCR, and the ratio of chemokine/GAPDH was considered to represent the final chemokine level. The plasma level of was measured by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe mean level of Mig mRNA in PBMCs of the patients with chronic hepatitis B was 0.6883-/+0.0693, which was significantly higher than that in normal controls (P<0.001). The plasma Mig level in the patients was 609.6-/+73.8 pg/ml, also significantly higher than that in normal controls (P<0.05). In patients with chronic hepatitis B, the level of Mig mRNA in the PBMCs was significantly correlated with plasma Mig level (r=0.7157, P<0.001), and plasma Mig level was correlated with plasma ALT level (r=0.7220, P<0.001) and plasma HBV DNA level (r=0.7266, P<0.001).

CONCLUSIONBoth the expression of Mig mRNA in PBMCs and plasma Mig concentration are elevated in patients with chronic hepatitis B. Mig plays an important role in migration of the inflammatory cells to the liver and mediates the development of chronic hepatitis B.

Adolescent ; Adult ; Chemokine CXCL9 ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B, Chronic ; blood ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction