Danshen, the dried roots and rhizomes of Salvia miltiorrhiza Bunge (S. miltiorrhiza), is widely used in the treatment of cardiovascular and cerebrovascular diseases. Tanshinones, the bioactive compounds from Danshen, exhibit a wide spectrum of pharmacological properties, suggesting their potential for future therapeutic applications. Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized, most of which belong to the CYP76 and CYP71 families. In this study, CYP81C16, a member of the CYP71 clan, was identified in S. miltiorrhiza. An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones, namely neocryptotanshinone, deoxyneocryptotanshinone, and danshenxinkuns A and B. SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%. Kinetic evaluations andin vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates. The overexpression of SmCYP81C16 promoted the accumulation of (iso)tanshinone in hairy root lines. The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones, either through microbial or plant metabolic engineering.
Humans ; Salvia miltiorrhiza/metabolism* ; Biosynthetic Pathways ; Quinones/metabolism* ; Plant Roots/metabolism* ; Gene Expression Regulation, Plant

OBJECTIVE：To observe the ef fectiveness and safety of bosentan in the treatment of hypoxic pulmonary hypertension（HPH）in neonates. METHODS ：From Jan. 2014 to Mar. 2019，a total of 82 HPH neonates hospitalized in the department of neonatology of our hospital were selected as research subjects. According to whether or not receiving bosentan therapy，50 cases were included into bosentan group and 32 cases into non-bosentan group. Meanwhile ，another 25 non-HPH neonates with serum sample retention time and general information such as gestational age at birth and day age matching the HPH group were selected as the control group. All neonates with HPH were given continuous intravenous infusion of Dopamine hydrochloride injection 5 mg/（kg·min）until PASP was normal. On this basis, neonates in the bosentan group were additionally given Bosentan tablets 1 mg/kg（fed after dissolving with appropriate amount of water for injection ）for q 12 h，72 h. The relationship between serum ET- 1 levels of neonates with HPH and PASP was analyzed ，as well as PASP before and after treatment and therapeutic efficacy between bosentan and non-bosentan groups ，the changes of arterial blood gas indexes and ADR in 3 groups were compared. RESULTS ：Before treatment ，the serum ET- 1 levels of bosentan group was （164.3±115.3）pg/mL，which was significantly higher than （41.9±3.7）pg/mL of control group and positively correlated with PASP level （r＝0.864，P＜0.001）. Total response rate of bosentan group was 90.00％，which was significantly higher than 71.88％ of non-bosentan group （P＜0.05）. After 72 h of treatment ，PASP of 2 groups was decreased significantly ，compared with before treatment （P＜0.001），and the bosentan group was significantly lower than the non-bosentan group （P＜0.05）. The PaO 2，SaO2，PaCO2 and OI in 3 groups was significantly improved compared with that before treatment （P＜0.001），and the PaO 2，SaO2 and OI in the bosentan group was significantly higher than that in the non-bosentan group （P＜0.05）. During the treatment period of bosentan and within one week after drug withdrawal，there was no significant change in serum LDH ，AST，ALT and Scr levels in neonates. There was no statistically significant difference in the incidence of feeding intolerance ，anemia，reduced WBC and reduced PLT in 3 groups（P＞0.05）. CONCLUSIONS： Bosentan can improve the oxygenation status of neonates with HPH, reduce PA SP，and short-termmedication is safe. com

Objective:To explore the application and influence of task-driven teaching in the internship of rehabilitation medicine among general medical students.Methods:A total of 256 students of the same grade from the fifth clinical college of Chongqing Medical University were included in this study. 128 students were selected as the experimental group to receive task-driven teaching, and another 128 students were selected as the control group to receive traditional teaching. Course design for students in the experimental group was evaluated, and the learning effects were compared between the two groups.Results:In the experimental group, more than 88% students were relatively satisfied with the tasks designed. The ability of self-study and independent thinking of students in the experimental groups was better than those in the control group ( P<0.05). In the aspect of problem solving, expression, cooperation, modern information utilization and creative thinking ability, students in the experimental group did a significantly better performance than those in the control group ( P=0.000), and students' learning motivation was significantly increased in the experimental group than in the control group ( P=0.000). Conclusion:In the internship of rehabilitation medicine, task-driven teaching was highly accepted among students and received fair satisfaction. It improves the learning effect of students majoring in general medicine and promotes the understanding for rehabilitation medicine, which lays a foundation for the work of primary hospitals in the future.

Objective To analyze the changes of serum hypoxia-inducible factor-1α(HIF-1α),vas-cular endothelial growth factor ( VEGF) and endothelin-1 ( ET-1) levels in neonates with hypoxia-induced pulmonary hypertension(HPH),and to explore the roles of three factors in the pathogenesis of HPH in neo-nates. Methods A total of 50 neonates with HPH in the neonatal intensive care unit as HPH group and an-other 25 non-HPH hospitalized neonates with similar clinical data in the same period as control group were enrolled from January 2014 to December 2017. The levels of serum HIF-1α,VEGF and ET-1 were deter-mined by enzyme linked immunosorbent assay when neonates were diagnosed as HPH and their pulmonary artery systolic blood pressure(PASP) decreased to 35 mmHg below respectively. The changes of three factors levels were analyzed and compared with those of control group at the same time point. Results (1) The levels of serum HIF-1α,VEGF and ET-1 of HPH group were significantly higher than those of the control group (F value were 151. 97,43. 31,and 129. 56 respectively,all P<0. 01). Furthermore,the more serious the grade of HPH,the higher the levels of three factors. The differences were statistically significant (all P<0. 01). (2) After PASP of neonates in HPH group decreased to 35 mmHg or below,the levels of serum HIF-1α,VEGF and ET-1 also significantly decreased,and the differences were statistically significant ( all P <0. 01). However,there were no statistically significant differences compared with those of the control group for the levels of serum HIF-1α and VEGF(both P>0. 05) in addition to serum ET-1 levels (F＝14. 98,P<0. 05). Conclusion High levels of HIF-1α,VEGF and ET-1 caused by hypoxia may play an important role in neonatal HPH.

OBJECTIVE:To investigate the omeprazole off-label use and provide reference for rational drug use in clinic. METHODS:In retrospective survey,1838 omeprazole prescriptions were selected from outpatient department during Jun. 2014 to May 2017 by random number table. According to the latest drug instruction,whether the off-label use or not was determined. Off-label use was determined by reviewing domestic and foreign guidelines,literatures and Micromedex database. RESULTS:Among 1838 pre-scriptions,there were 1750 prescriptions of off-label drug use (95.21％),involving 13 items and 3 types of off-label drug use. Main type of off-label drug use was over-indication medication,involving 1747 prescriptions (96.47％). There was no statistical significance in the incidence of off-label drug use between digestive department and non-digestive department or among physicians at different levels(P>0.05). Among 13 items of off-label drug use,7 items were supported by evidence-based medicine evidence of domestic and foreign guideline,expert consensus/suggestion or literature reports,etc.;among which 1 item of off-label drug use were included in Micromedex grading system and other 6 items had no evidence-based medicine evidence. CONCLUSIONS:The phenomenon of omeprazole off-label use is widespread in outpatient department of the hospital,and some off-label drug use are sup-ported by evidences. There are differences in the quality of those evidences. It is suggested to standardize the off-label drug use to avoid legal risks and guarantee the safety of drug use for patients.

Objective To better understand the clinical characteristics of Familial Idiopathic Basal Ganglia Calcifi?cation (FIBGC), including at the perspective of hereditary pattern, clinical test results, onset age, clinical heterogeneity and the volume of basal ganglia calcification (VBGC). Method 8 Eight FIBGC families were collected and draw family pedigrees were draw. Analysis of was conducted on the patient's clinical test results, head CT and MRI changes, onset ag?es, relationship of clinical manifestations with VBGC. Results No significant difference was found in serum calcium, alu?minum, arsenic, cobalt, magnesium, phosphorus, iron, parathyroid hormone and calcitonin concentration between the fam?ily members of patients and healthy controls (P>0.05). Family members from 8 FIBGC families including the two with consanguineous marriage manifested autosomal dominant heredity. The severity of , symptomatic s was correlated with VBGCpatients showed the same clinical manifestations in the dyskinesia family. The psychiatric symptoms was not asso? ciated with VBGC whereas patients with dyskinesia had a large VBGC. There was a significant difference in onset age be?tween patients with psychiatric symptoms and those with dyskinesia. P.atients with dyskinesia suffer larger VBGC, and is characterized by Patients with dyskinesia had relatively later onset age (43.95 ± 2.47 y) whereas those with. psychiatric symptoms hadsymptomatic patients with early onset age (31.32±10.16y). The comparison of the onset age (43.954±2.473 vs. 31.319±10.156 y, t=4.438, P=0.001) and VBGC (1.748±0.622 vs. 0.392±0.276 cm3, t=2.518, P=0.028) with symptom?atic patients between dyskinesia and psychogenic families was significant. Conclusions Eight FIBGC families manifested autosomal dominant heredity. Patients with dyskinesia suffer have a larger VBGC and are associated with a, and is char?acterized by relatively later onset age. In contrast, patients with psychiatric symptomspsychogeny is not related withhave a the small VBGC and showedand their age of onset is young. earlier onset age.

Objective To understand the characteristics of pathogenic bacteria and drug resistance of ventilator-associated pneu-monia(VAP)in neonatal intensive care unit (NICU),and to explore the corresponding prevention and control measures,and to pro-vide the basis for the VAP antibiotic treatment.Methods A total of 80 children with respiratory failure and ventilator assisted breathing were selected from the NICU as objects in this study.The clinical data,pathogenic bacteria and drug resistance were ana-lyzed retrospectively.Results In 80 cases,the incidence of VAP was 43.75% (35/80),a total of 70 strains of pathogenic bacteria were isolated,gram negative bacilli accounted for the highest proportion,accounting for 81.43% (57/70).Pseudomonas aeruginosa, Klebsiella pneumoniae were the most common gram negative bacilli.Gram positive cocci accounted for 8.57% (13/70),which domi-nated by methicillin resistant Staphylococcus aureus,in addition to susceptible to vancomycin,but resistant to the other antibiotics. Conclusion Gram negative bacillus are the main bacteria in VAP cases,and which are multiple drug-resistant pathogens.

Objective To knockout and identify the Antigen 43 (Ag43) in the Escherichia Coli JM109 .Methods Mutation group Ⅱ introns RNA protein complexes (RNP) gene sequence was obtained by Sigma Company′s TargeTron Gene Knockout Sys‐tem and Ag43 gene specific designed PCR primers amplification ,then ,to acquired Ag43 specific recombinant RNP plasmid pACD4K‐Ag4 ,this gene sequence was inserted into the plasmid pACD4K‐C of RNA′s expression .Finally ,pEGFP‐Ag43 was trans‐formed into JM109 and inserted the group Ⅱ intron into the Ag43′s locus by IPTG inducing expression .Results The best insertion locus was between 1 812 and 1 913 .Through the agarose electrophoresis gel ,the RNP gene sequence was consistent with the expec‐ted value (350 bp) .The pEGFP‐Ag43 vector was correctly constructed which was proofed by endonuclease Nhe Ⅰ and Hind ⅡI di‐gestion as predicted products (3 646 and 4 029 bp;7 000 and 550 bp ,respectively ) .The PCR and gene sequence results indicated that the group Ⅱ intron was inserted into the locus between 1 812 and 1 913 in the Ag43 gene .Conclusion Successful knockout of the Ag43 in Escherichia Coli JM109 found basis to further study the Ag43′s function and regard the coli as host bacteria of Ag43 chimeric protein recombinant .

OBJECTIVETo study the mechanism of secondary metabolites of some phenolic acids in the hairy roots of Salvia miltiorrhiza induced by methyl jasmonate.

METHODThe hairy roots of S. miltiorrhiza were induced with methyl jasmonate (100 micromol x L(-1)) and collected at 0, 12, 24, 36 h after treatment. Real-time quantitative PCR was used for detecting the mRNA expression level of the key enzyme genes on the secondary metabolites pathway of rosmarinic acid, while a LC-MS method was developed to determine the content of rosmarinic acid, caffeic acid and salvianolic acid B.

RESULT AND CONCLUSIONThe concentration of phenolic acids grew up and accumulated quickly in the hairy roots with exogenous signal molecule MJ induced, and it was showed that the content of CA and RA reached the maximum after 24 h and the content of LAB reached the maximum in 36 h by MJ induced. The induction mechanism may be activated with different levels of RA synthesis in PAL, 4CL, C4H genes on the key enzyme phenylalanine pathway and TAT, HPPR genes on tyrosine pathway. The time of gene expression was different, among them, 4CL and PAL genes were more important. In a word, the result can provide some basis data about the mechanism of secondary metabolites of phenolic acids for further research.

Cyclopentanes ; analysis ; metabolism ; Gene Expression Regulation, Plant ; Hydroxybenzoates ; analysis ; metabolism ; Oxylipins ; analysis ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; enzymology ; genetics ; metabolism