DNA barcoding of Gentiana crassicaulis in Lijiang and evaluation of root processing methods / 药学学报

The key factors for producing the best quality Chinese herbal medicines are high-quality germplasm, suitable cultivation area and the proper processing methods for herbal raw materials. Gentiana crassicaulis in Gentiana (Sect. Cruciata), Gentianaceae is one of the original plants of the Chinese herb Qinjiao (Gentianae Macrophyllae Radix), and its type specimen was collected in Lijiang, Yunnan. There is a long planting history of the herb in this area. In this study a sampling plot was designated in these traditional planting areas. G. crassicaulis was planted and herbal raw materials were harvested from the plot. The raw materials were prepared locally and at a pharmaceutical factory in Shanghai using processing methods such as "sweating" or "no sweating", "slicing" or "no slicing" (whole root), and "stoving" or "no stoving" (air drying). The quality of all processed samples was evaluated. In addition, molecular markers were determined for identifying cultivated and wild samples from Lijiang, Yunnan. The results are as follows: ① Samples from the sampling plot and the field are taxonomically identified as Gentiana crassicaulis. ② A total of 270 sequences of trnC-GCA-petN, atpB-rbcL, psbN, ndhB-rps7 and ycf1 were obtained, and three genotypes were determined from the cultivated samples; the type III was shared by both cultivated and wild plants. Based on the molecular markers, a DNA barcoding method to identify cultivated and wild samples of G. crassicaulis from Lijiang, Yunnan was established. ③ Total content of loganic acid and gentiopicroside in all samples was ≥ 2.5%, and above the Chinese Pharmacopoeia (2020) limit. ④ In HPLC fingerprinting, 9 common peaks were assigned and similarity between all samples was > 0.999; and ⑤ In a PCA score plot all slice samples were clustered, while whole root samples were scattered. Therefore, our studies could provide basic data for optimizing the processing method, producing best quality Gentianae Macrophyllae Radix, and evaluating the quality of different ecotype varieties and the multiple origin of herbal medicines.

Transcriptome analysis and validation of key genes involved in biosynthesis of iridoids in Gentiana lhassica / 中国中药杂志

As the main chemical constituents, iridoids are widely distributed within Gentiana, Gentianaceae, with promising bioactivities. Based on the previous work, the transcriptome of G. lhassica, an original plant of Tibetan herb "Jieji Nabao", was sequenced and analyzed in this study, and the transcriptome databases of roots, stems, leaves, and flowers were constructed so as to explore unigenes that may encode the key enzymes in the biosynthetic pathway of iridoids. Then, qRT-PCR was used to validate the relative expression levels of 11 genes named AACT, DXS, MCS, HDS, IDI, GPPS, GES, G10H, 7-DLNGT, 7-DLGT, and SLS in roots, stems, leaves, and flowers. Also, the total contents of gentiopicroside and loganic acid were determined by HPLC, respectively. The results are as follows:(1)a total of 76 486 unigenes with an average length of 852 bp were obtained;(2)335 unigenes were involved in 19 stan-dard secondary metabolism pathways in KEGG database, with phenylpropanoid biosynthesis having the maximum number(75 unigenes), and no isoflavone biosynthetic pathway was annotated;(3)171 unigenes participatedin 27 key enzymes encoding in the biosynthetic pathway of iridoids, and 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR) gene was highly expressed;(4)qRT-PCR results were approximately consistent with RNA-Seq data and the relative expression levels of the 11 genes were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root);(5)the total contents of gentiopicroside and loganic acid were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root), and the difference was significant. This study provides basic scientific data for accurate species identification, evaluation of germplasm resources, research on secondary pro-duct accumulation of medicinal plants within Gentianaceae, and protection of endangered alpine species.

Transcriptome analysis and exploration of genes involved in the biosynthesis of iridoids in Gentiana crassicaulis (Gentianaceae) / 药学学报

italic>Gentiana crassicaulis Duthie ex Burk. in Gentiana (Sect. Cruciata), Gentianaceae, is one of the original plants of both Gentianae Macrophyllae Radix and Tibetan herb Jie-Ji Na-Bao, which contain such bioactive iridoids as gentiopicroside, loganic acid and others. In this study, based on previous work, the transcriptome of G. crassicaulis was sequenced and analyzed to construct transcriptome databases of roots, stems, leaves and flowers. qRT-PCR verification was conducted for parts of unigenes that may be key enzymes in the pathway of iridoid biosynthesis. The results are as follows: ① a total of 159 534 unigenes were obtained, with an average length of 679 bp. According to the functional classification of GO, unigenes can be divided into 3 categories with 67 branches. The unigenes were aligned in the KOG database and were classified into 25 categories according to function. ② In the KEGG database, 215 unigenes were implicated in 20 standard secondary metabolism pathways. The analysis shows that 305 unigenes encoded 28 key enzymes in the pathway of iridoid biosynthesis, and their expression in different organs is different; and ③ qRT-PCR was approximately consistent with RNA-Seq results. The 7 annotated unigenes identified in this study, HMGS, DXS, MCS, GPPS, G10H, 7-DLNGT and STR, all had higher relative expression levels in the above-ground parts (stem, leaf and flower) than in the underground part (root). Iridoids are common active and index components of such traditional Chinese medicines as Qinjiao, Longdan, Dangyao, and Qingyedan, among others. Therefore, this work provides basic scientific data for further development including obtaining active components or intermediates through biotechnology, exploring the accumulation of effective components, evaluating the quality of different ecotype varieties, and identifying authentic biosynthesis pathways of medicinal materials.

Glutathione Modified Ordered Mesoporous Carbon for Separation and Preconcentration of Trace Cadmium / 分析化学

A new derivative of ordered mesoporous carbon (GSH-CMK-3) has been prepared by chemical modification and functionalization of ordered mesoporous carbon with glutathione. The composite was fully characterized by means of Fourier transforming infrared spectroscopy (FT-IR), scanning electron microscope (SEM) image and thermogravimetric analysis (TGA). GSH-CMK-3 showed favorable chemical stability, thermal stability and excellent adsorption performance toward Cd2+and a maximum adsorption capacity of 87.87 mg/g was achieved. The adsorption kinetic follows pseudo-second-order adsorption model. In a sequential injection mini-column separation system, 0. 006 mol/L sulfourea (in 0. 2 mol/L HNO3) was adopted as eluent and the elution efficiency was 96%. With a sample volume of 1000 μL and 50 μL of eluate, an enrichment factor of 17.3 was obtained. A procedure for on-line selective separation and preconcentration of trace Cd2+was developed by graphite furnace atomic absorption spectrometry. Along with a detection limit of 1.9 ng/L (n=7,3σ) and a linear range of 0.05-0.20 μg/L, a RSD of 2.9% (n=13,100 ng/L) was achieved. The method was validated by analyzing Cd2+in a certified reference material GBW08608.

Scientific experiment exploration course of biomedical engineering specialty / 医疗卫生装备

Objective To study the teaching innovation of the scientific experiment exploration course in biomedical engineering specialty based on its characteristics involving in animal physiological parameters acquisition, circuit system design, signal processing and etc. Methods The course aimed to guide the student in observation learning, practice and inquiry with considerations on course significance,basic principles for designing the course,implementation scheme of course teaching, suggestions for course improvement, instances of practical teaching and etc. Results Four basic principles were proposed for course teaching. An implementation scheme was determined with emphases on teaching materials, theory, practice, discussion and exploration. The course had the rule being inquiry, and some suggestions were put forward about student feedback, scientific research progress, experiment platform renewal and etc. Conclusion Teaching practice proves that the improved course contributes to enhancing the student's understanding of experiment exploration of biomedical engineering specialty and to promoting multi-discipline fusion and cross-over personnel training, and thus is of great reference for other universities.

Comparison of clinical efficacy of heated humidified high flow nasal cannula versus nasal continuous positive airway pressure in treatment of respiratory distress syndrome in very low birth weight infants / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To compare the differences of clinical efficacy between heated humidified high-flow nasal cannula (HHHFNC) ventilation and nasal continuous positive airway pressure (NCPAP) in the treatment of respiratory distress syndrome (RDS) in very low birth weight (VLBW) infants.</p><p><b>METHODS</b>A total of 66 VLBW infants who were admitted to the neonatal intensive care unit were diagnosed with RDS, and they were randomly assigned to HHHFNC group and NCPAP group after receiving treatment with porcine pulmonary surfactant and conventional treatment. The changes in clinical symptoms and the incidence of complications were observed in the two groups.</p><p><b>RESULTS</b>The HHHFN group had significantly earlier first milk feeding and full enteral feeding, significantly shorter oxygen exposure time and invasive ventilation time, and significantly lower incidences of second intubation within 7 days, nasal injury, air leak, and abdominal distention, as compared with the NCPAP group.</p><p><b>CONCLUSIONS</b>Compared with NCPAP, HHHFNC causes slighter injury and has better tolerability, and it can be considered as the first choice of noninvasive ventilation in the treatment of RDS in VLBW infants.</p>