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1.
Acta Pharmaceutica Sinica ; (12): 673-677, 2024.
Article in Chinese | WPRIM | ID: wpr-1016628

ABSTRACT

Five flavonoid glycosides were isolated from the methanol and ethyl acetate fractions of the ethanol extract of Diphylleia sinensi by using various chromatographic methods, including silica gel, MCI gel, Sephadex LH-20, ODS and semi-preparative HPLC. The structures of the isolated compounds were identified as diphyflavonoid A (1), diphyflavonoid B (2), quercetin-3-O-β-D-glucopyranoside (3), kaempferol-3-O-β-D-glucopyranoside (4), kaempferol-3-O-(6″-O-acetyl)-β-D-glucopyranoside (5) by spectroscopy methods (1D NMR, 2D NMR, UV, IR, and MS). Compounds 1 and 2 were two new flavonoid glycosides, and compounds 3 and 5 were isolated from the genus Diphylleia for the first time.

2.
Acta Pharmaceutica Sinica ; (12): 3296-3310, 2023.
Article in Chinese | WPRIM | ID: wpr-999078

ABSTRACT

Quantitative systems pharmacology (QSP) modeling is an emerging computational medicine approach with growing applications and significance in modern drug development. QSP models are generally formulated based on multiscale disease mechanisms and drug-target interactions, which makes them capable of integrating multimodal data from the preclinical and clinical space. This also enables them to generate quantitative characterization of the dynamic disease progression as well as high-throughput predictions of drug-induced efficacy and toxicity signals. Therefore, QSP modeling and model-based virtual clinical trials have been widely implemented to guide drug development, in scenarios such as target identification and assessment, clinical trial design, evaluation of combination therapy and biomarkers, and personalized medicine. In US and Europe, QSP modeling has been developing rapidly in the past 10 years and is now an integral part of the model-informed drug development paradigm; however, in China it is still a nascent field. Here we will present a comprehensive review of the recent advancements of QSP and its impact in modern drug development through a number of case studies. This review will provide guidance for the future drug development efforts and the growth of QSP practice in China.

3.
Shanghai Journal of Preventive Medicine ; (12): 301-303, 2023.
Article in Chinese | WPRIM | ID: wpr-976260

ABSTRACT

A patient with SARS-CoV-2 infection was adimitted to Shanghai Shibei hospital of Jing'an District in early 2023. According to the patient's complaits, clinical manifestations, physical symptoms, laboratory examination, radiological image results, plus lumbar puncture, the patient was diagnosed with novel coronavirus encephalitis. The patient was discharged from the hospital after a combined treatment of Chinese and western medicine.

4.
Protein & Cell ; (12): 105-122, 2023.
Article in English | WPRIM | ID: wpr-971612

ABSTRACT

Glioblastoma multiforme (GBM), a highly malignant and heterogeneous brain tumor, contains various types of tumor and non-tumor cells. Whether GBM cells can trans-differentiate into non-neural cell types, including mural cells or endothelial cells (ECs), to support tumor growth and invasion remains controversial. Here we generated two genetic GBM models de novo in immunocompetent mouse brains, mimicking essential pathological and molecular features of human GBMs. Lineage-tracing and transplantation studies demonstrated that, although blood vessels in GBM brains underwent drastic remodeling, evidence of trans-differentiation of GBM cells into vascular cells was barely detected. Intriguingly, GBM cells could promiscuously express markers for mural cells during gliomagenesis. Furthermore, single-cell RNA sequencing showed that patterns of copy number variations (CNVs) of mural cells and ECs were distinct from those of GBM cells, indicating discrete origins of GBM cells and vascular components. Importantly, single-cell CNV analysis of human GBM specimens also suggested that GBM cells and vascular cells are likely separate lineages. Rather than expansion owing to trans-differentiation, vascular cell expanded by proliferation during tumorigenesis. Therefore, cross-lineage trans-differentiation of GBM cells is very unlikely to occur during gliomagenesis. Our findings advance understanding of cell lineage dynamics during gliomagenesis, and have implications for targeted treatment of GBMs.


Subject(s)
Mice , Animals , Humans , Glioblastoma/pathology , Endothelial Cells/pathology , DNA Copy Number Variations , Brain/metabolism , Brain Neoplasms/pathology
5.
China Journal of Chinese Materia Medica ; (24): 3730-3735, 2023.
Article in Chinese | WPRIM | ID: wpr-981505

ABSTRACT

Artemisia stolonifera is a relative of A. argyi. The two species are difficult to be distinguished due to the similarity in leaf shape and have even less distinctive features after processing. This study aims to establish a method to quickly distinguish between them. At the same time, we examined the reasonability and applicability of the specific polymerase chain reaction(PCR) method. The C/T single nucleotide polymorphism was detected at the position 202 of the sequence, based on which specific primers were designed to identify these two species. The PCR with the specific primer JNC-F and the universal primer ITS3R produced a specific band at 218 bp for A. argyi and no band for A. stolonifera, which can be used to detect at least 3% of A. argyi samples mixed in A. stolonifera samples. The PCR with the specific primer KY-F and the universal primer ITS3R produced a specific band at 218 bp for A. stolonifera and no band for A. argyi, which can be used to detect at least 5% of A. stolonifera samples mixed with A. argyi. The limit of detection of the established method was 5 ng DNA. The established PCR method can accurately distinguish between A. stolonifera and A. argyi, which provides an experimental basis for the quality control of A. stolonifera and determines whether the herbs are adulterated.


Subject(s)
Artemisia/genetics , Trichomes , Polymerase Chain Reaction , Nucleic Acid Amplification Techniques , Plant Leaves/genetics
6.
China Journal of Chinese Materia Medica ; (24): 3722-3729, 2023.
Article in Chinese | WPRIM | ID: wpr-981504

ABSTRACT

This paper aims to compare the difference of growth and quality between wild and cultivated Artemisia stolonifera, thereby providing references for further development and utilization of A. stolonifera. The wild and cultivated A. stolonifera from different altitudes were collected, and the agronomic characters, moxa yield, volatile components, flavonoids, and phenolic acids were determined. The results showed that the cultivated species were taller and stronger, with more leaves and branches, than the wild species. The moxa yield and combustion quality of wild products were higher than those of cultivated products. The content of main volatile components in cultivated products was higher than that in wild products. The content of flavonoids and phenolic acids in wild products was higher than that in cultivated products. At high altitude, the ignition performance, combustion persistence, comprehensive combustion performance, and heat release during combustion of the wild and cultivated A. stolonifera. were optimal. At middle altitude, the content of main characteristic volatile components and flavone phenolic acids in the leaves of the cultivated and wild A. stolonifera were the highest. At low altitude, the combustion quality and the content of the above components of the cultivated A. stolonifera decrease significantly. Considering the combustion quality and the content of the internal components of the leaf lint, the middle and high altitude areas are suitable for the artificial cultivation of A. stolonifera.


Subject(s)
Artemisia , Agriculture , Flavonoids , Plant Leaves , Drugs, Chinese Herbal
7.
China Journal of Chinese Materia Medica ; (24): 3715-3721, 2023.
Article in Chinese | WPRIM | ID: wpr-981503

ABSTRACT

The purpose of this study was to analyze the effects of shading intensity on the growth, yield, and quality of Artemisia stolonifera so as to provide references for the artificial cultivation of A. stolonifera. The seedlings of A. stolonifera with consistent growth underwent shading treatment at four shading intensity levels(0, 55%, 85%, and 95%) with different layers of black shading nets. The agronomic indexes, yield, moxa yield, total ash, quality characteristics of moxa during combustion and pyrolysis, main volatile components, flavonoids, and phenolic acids were measured. The results showed that under shading conditions, the stem diameter, leaf width, 5-leaf spacing, branch number, and yield of A. stolonifera decreased significantly, while the plant height, leaf length, leaf number, chlorophyll content, and moxa yield increased first and then decreased with the increase in shading intensity. The burning performance of moxa under natural light was better than that under moderate and severe shading conditions. The content of eucalyptol first increased and then decreased with the increase in shading intensity. The humulene content was negatively correlated with shading intensity. Other major volatile components showed no significant difference under various shading conditions. The content of neochlorogenic acid, cryptochlorogenic acid, isoschaftoside, and isochlorogenic acid B was positively correlated with shading intensity, while the content of chlorogenic acid, isochlorogenic acid A, and isochlorogenic acid C decreased first and then increased with the increase in shading intensity. To sum up, A. stolonifera is a light-loving plant, and shading can greatly reduce the yield, the content of internal components, and the burning performance of moxa. It is the main reason why A. stolonifera is mainly distributed in the forest edge, open forest, roadside, and wasteland grass in the middle and high mountains in the wild. For artificial domestication and cultivation of A. stolonifera, it is better to select plots with sufficient light.

8.
China Journal of Chinese Materia Medica ; (24): 3421-3439, 2023.
Article in Chinese | WPRIM | ID: wpr-981478

ABSTRACT

Chinese medicinal resources are the material basis for the survival and development of traditional Chinese medicine(TCM)and the sustainable development of Chinese medicinal resources is also an important project for the modernization of TCM in China. With the increasing demand for Chinese medicinal resources in China, over-exploitation has destroyed Chinese medicinal resources, resulting in a shortage of many natural medicinal resources in China and making the sustainable development of TCM in trouble. The introduced new foreign medicinal resources have become effective supplement and replacement for Chinese medicinal resources to some extent. However, the development and utilization of new foreign medicinal resources in China are different. To fully understand the development of new foreign medicinal resources in China, this paper, taking 43 new foreign medicinal resources such as Acacia nilotica as objects, sorted out the introduction forms and policies of new foreign medicinal resources, overviewed its current development status in China, summarized the application experience of new foreign medicinal resources in the place of origin, as well as the research progress and problems of new foreign medicinal resources in China and abroad, and analyzed the research situation, which can enrich Chinese medicinal resources and other uses, promote the sustainable development of Chinese medicinal resources, and provide ideas for further development and research of new foreign medicinal resources.


Subject(s)
Drugs, Chinese Herbal/therapeutic use , Medicine, Chinese Traditional , Conservation of Natural Resources , Sustainable Development , Internationality , China
9.
China Journal of Chinese Materia Medica ; (24): 2160-2185, 2023.
Article in Chinese | WPRIM | ID: wpr-981347

ABSTRACT

Starting with the relationship between mulberry leaves and silkworm droppings as food and metabolites, this study systematically compared the chemical components, screened out differential components, and quantitatively analyzed the main differential components based on ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and UPLC-Q-TRAP-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Moreover, the in vitro enzymatic transformation of the representative differential components was studied. The results showed that(1) 95 components were identified from mulberry leaves and silkworm droppings, among which 27 components only exist in mulberry leaves and 8 components in silkworm droppings. The main differential components were flavonoid glycosides and chlorogenic acids.(2) Nineteen components with significant difference were quantitatively analyzed, and the components with significant differences and high content were neochlorogenic acid, chlorogenic acid, and rutin.(3) The crude protease in the mid-gut of silkworm significantly metabolized neochlorogenic acid and chlorogenic acid, which may be an important reason for the efficacy change in mulberry leaves and silkworm droppings. This study lays a scientific foundation for the development, utilization, and quality control of mulberry leaves and silkworm droppings. It provides references for clarifying the possible material basis and mechanism of the pungent-cool and dispersing nature of mulberry leaves transforming into the pungent-warm and dampness-resolving nature of silkworm droppings, and offers a new idea for the study of nature-effect transformation mechanism of traditional Chinese medicine.


Subject(s)
Animals , Bombyx , Morus/chemistry , Chlorogenic Acid/analysis , Gas Chromatography-Mass Spectrometry , Chromatography, High Pressure Liquid/methods , Plant Leaves/chemistry
10.
China Tropical Medicine ; (12): 176-2023.
Article in Chinese | WPRIM | ID: wpr-979613

ABSTRACT

@#Abstract: Objective To analyze the antimicrobial resistance rate and risk factors of multi drug resistant organisms (MDRO) in bloodstream infection for rational treatment. Methods A total of 696 cases of bloodstream infections of Staphylococcus aureus, Enterococcus, Enterobacteriaceae (excluding Salmonella and Shigella), Pseudomonas aeruginosa and Acinetobacter in our hospital from 2017 to 2021 were selected, and 711 pathogenic strains were isolated from their whole blood samples. The antimicrobial resistance rates of various multi drug resistant strains were analyzed and the risk factors of MDRO infection were analyzed. Results 696 non repeated cases were screened out from 13 187 whole blood culture specimens, with a positive rate of 5.3%, and a total of 711 blood influenza pathogens were detected, among them, 350 strains of MDRO were detected with a detection rate of 49.23% (350/711). Among the pathogenic bacteria of bloodstream infection, Escherichia coli was the most, with 277 strains accounting for 38.96% (277/711), of which 201 strains were MDRO, accounting for 57.43% (201/350); followed by Klebsiella pneumoniae and Staphylococcus aureus, with 155 strains accounting for 21.80% (155/711) and 89 strains accounting for 12.52% (89/711), among which 43 strains of Klebsiella pneumoniae MDRO accounted for 12.29% (43/350) and 38 strains of Staphylococcus aureus MDRO accounted for 10.86% (38/350). The change trend of the three pathogens during 2017-2021 was not obvious. The drug sensitivity test showed that Escherichia coli and Klebsiella pneumoniae were highly resistant to cephalosporins and fluoroquinolones, and the drug resistance rate of aminoglycosides was relatively low. They had almost no resistance to cephalosporins and carbapenems. Staphylococcus aureus has a high resistance rate to lincomycin and macrolides, but no resistance to oxazolidinone, glycopeptides and glycylcyclins. There were 350 cases of MDRO infection and 361 cases of non MDRO infection. Univariate analysis showed that the age, sex, cardiovascular and cerebrovascular history, renal insufficiency, lung disease, hypoalbuminemia, hepatobiliary disease, electrolyte disorder and anemia of the patients had no statistical significance in MDRO infection (P>0.05); diabetes, urinary tract infection, surgical operation and burn were the influencing factors of MDRO (P<0.05). According to logistic analysis, diabetes, urinary tract infection, surgical operation and burn were the risk factors of MDRO infection (P<0.05). Conclusion The infection of MDRO in patients with bloodstream infection is serious, and early prevention and control should be paid attention to, and the principle of graded use of antibiotics should be strictly observed, and the rational application should be carried out to actively and effectively control the production of MDRO.

11.
Chinese Acupuncture & Moxibustion ; (12): 317-321, 2023.
Article in Chinese | WPRIM | ID: wpr-969991

ABSTRACT

OBJECTIVE@#To observe the effects of moxibustion on the ultrastructure of synovial cells of knee joint and serum cytokines in adjuvant arthritis (AA) rats, and to explore the potential mechanism of moxibustion in treatment of rheumatoid arthritis.@*METHODS@#Forty-five Wistar male rats were randomly divided into a normal group, a model group and a moxibustion group, with 15 rats in each group. In the model group and the moxibustion group, the AA model was replicated under wind, cold and humid environment and by injection with complete freund's adjuvant. In the moxibustion group, moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) was used, 20 min each time, once daily, for consecutive 21 days. In the normal group and the model group, no intervention was processed. The scores of the knee joint swelling degree (JSD) and arthritis index (AI) were compared among groups. The ultrastructure of synovial cells of knee joint were observed under transmission electron microscope (TEM). The levels of serum cytokines such as tumor necrosis factor-α (TNF-α), interieukin (IL)-1β, IL-6 and IL-10 were detected using ELISA method.@*RESULTS@#Compared with the normal group, JSD and AI scores, the levels of TNF-α, IL-1β and IL-6 were increased (P<0.01), while IL-10 was reduced (P<0.01) in the model group after intervention. JSD and AI scores, and the levels of TNF-α, IL-1β and IL-6 were lower (P<0.05, P<0.01), while the level of IL-10 was higher (P<0.01) in the moxibustion group compared with the model group. Compared with the normal group, the ultrastructure of synovial cell was obviously damaged in the model group, and the damage was attenuated in the moxibustion group compared with the model group.@*CONCLUSION@#Moxibustion can reduce the symptoms of arthritis in AA rats, which may be related to the improvement of the ultrastructure of synovial cells and the regulation of cytokines.


Subject(s)
Male , Rats , Animals , Cytokines , Interleukin-10 , Arthritis, Experimental , Tumor Necrosis Factor-alpha , Interleukin-6 , Moxibustion , Rats, Wistar , Knee Joint
12.
Chinese Journal of Preventive Medicine ; (12): 268-272, 2023.
Article in Chinese | WPRIM | ID: wpr-969877

ABSTRACT

Objective: To establish a rapid and specific quantitative real-time PCR (qPCR) method for the detection of SARS-CoV-2 subgenomic nucleocapsid RNA (SgN) in patients with COVID-19 or environmental samples. Methods: The qPCR assay was established by designing specific primers and TaqMan probe based on the SARS-CoV-2 genomic sequence in Global Initiative of Sharing All Influenza Data (GISAID) database. The reaction conditions were optimized by using different annealing temperature, different primers and probe concentrations and the standard curve was established. Further, the specificity, sensitivity and repeatability were also assessed. The established SgN and genomic RNA (gRNA) qPCR assays were both applied to detect 21 environmental samples and 351 clinical samples containing 48 recovered patients. In the specimens with both positive gRNA and positive SgN, 25 specimens were inoculated on cells. Results: The primers and probes of SgN had good specificity for SARS-CoV-2. The minimum detection limit of the preliminarily established qPCR detection method for SgN was 1.5×102 copies/ml, with a coefficient of variation less than 1%. The positive rate of gRNA in 372 samples was 97.04% (361/372). The positive rates of SgN in positive environmental samples and positive clinical samples were 36.84% (7/19) and 49.42% (169/342), respectively. The positive rate and copy number of SgN in Wild strain were lower than those of SARS-CoV-2 Delta strain. Among the 25 SgN positive samples, 12 samples within 5 days of sampling time were all isolated with virus; 13 samples sampled for more than 12 days had no cytopathic effect. Conclusion: A qPCR method for the detection of SARS-CoV-2 SgN has been successfully established. The sensitivity, specificity and repeatability of this method are good.


Subject(s)
Humans , SARS-CoV-2/genetics , COVID-19/diagnosis , Subgenomic RNA , Real-Time Polymerase Chain Reaction/methods , RNA, Viral/genetics , Sensitivity and Specificity , Nucleocapsid/chemistry , COVID-19 Testing
13.
China Journal of Chinese Materia Medica ; (24): 4782-4788, 2023.
Article in Chinese | WPRIM | ID: wpr-1008645

ABSTRACT

A cross-sectional study method combined with two types of traditional Chinese medicine(TCM) syndrome differentiation methods was adopted to investigate the clinical symptoms and distribution characteristics of TCM syndromes in patients with pulmonary nodules from the perspectives of number, size, nature, and stability of pulmonary nodules by using the χ~2 test, systematic clustering and Apriori algorithm correlation analysis. The common clinical symptoms of pulmonary nodules were fatigue(77.35%) and irritability(75.40%), and 40 symptoms were clustered into 3 groups(digestive system symptoms, respiratory system symptoms, and emotional and systemic symptoms) and 8 major symptom categories. The proportion of cold and heat in complexity syndrome(63.43%) was higher based on cold-heat syndrome differentiation. The top two syndromes were Qi deficiency syndrome(88.03%) and Qi depression syndrome(83.17%) based on disease syndrome differentiation. Yang deficiency syndrome(60.52%) was more than Yin deficiency syndrome(50.16%). There were higher proportions of phlegm syndrome(78.67%) and Yang deficiency syndrome(69.33%) of so-litary pulmonary nodules in terms of the number of pulmonary nodules. In terms of size, the proportion of phlegm syndrome decreased as the mean diameter of pulmonary nodules increased, while the proportions of Yang deficiency syndrome and blood stasis syndrome increased. The distribution of Qi depression syndrome was more in those with mean diameter<10 mm(85.02%, P=0.044) and cold syndrome was more in those with mean diameter ≥10 mm(16.67%, P=0.024). In terms of the nature of pulmonary nodules, the proportions of Qi depression syndrome and heat syndrome decreased with the increase in solid components of pulmonary nodules, while the proportions of Yin deficiency syndrome and cold and heat in complexity syndrome increased. The blood stasis syndrome accounted for a higher proportion of pulmonary nodules with solid components. In terms of the stability of pulmonary nodules, dampness syndrome(72.97%), blood stasis syndrome(37.84%), and cold and heat in complexity syndrome(70.27%) accounted for higher proportions. In addition, patients with new nodules presented higher proportions in Qi inversion syndrome(52.00%, P=0.007) and cold and heat in complexity syndrome(66.00%, P=0.008). Meanwhile, 11 syndromes were associated and 4 common compound syndromes were obtained(Qi deficiency and depression syndrome, Qi depression and phlegm coagulation syndrome, Qi deficiency and phlegm coagulation syndrome, and Qi deficiency and dampness obstruction syndrome). Qi deficiency syndrome and Qi depression syndrome could be associated with other syndromes. The results show that the main clinical symptoms of pulmonary nodules are fatigue and irritability. The main TCM syndromes of pulmonary nodules are Qi deficiency syndrome, Qi depression syndrome, Yang deficiency syndrome, and cold and heat in complexity syndrome. The distribution of TCM syndromes is significantly correlated with the size of pulmonary nodules and the presence or absence of new nodules. The common compound syndromes are Qi deficiency and depression syndrome, Qi depression and phlegm coagulation syndrome, Qi deficiency and phlegm coagulation syndrome, and Qi deficiency and dampness obstruction syndrome.


Subject(s)
Humans , Medicine, Chinese Traditional , Yin Deficiency/diagnosis , Yang Deficiency/diagnosis , Cross-Sectional Studies , Syndrome
14.
Neuroscience Bulletin ; (6): 1039-1049, 2023.
Article in English | WPRIM | ID: wpr-982429

ABSTRACT

In this study, we explored the neural mechanism underlying impaired stereopsis and possible functional plasticity after strabismus surgery. We enrolled 18 stereo-deficient patients with intermittent exotropia before and after surgery, along with 18 healthy controls. Functional magnetic resonance imaging data were collected when participants viewed three-dimensional stimuli. Compared with controls, preoperative patients showed hypoactivation in higher-level dorsal (visual and parietal) areas and ventral visual areas. Pre- and postoperative activation did not significantly differ in patients overall; patients with improved stereopsis showed stronger postoperative activation than preoperative activation in the right V3A and left intraparietal sulcus. Worse stereopsis and fusional control were correlated with preoperative hypoactivation, suggesting that cortical deficits along the two streams might reflect impaired stereopsis in intermittent exotropia. The correlation between improved stereopsis and activation in the right V3A after surgery indicates that functional plasticity may underlie the improvement of stereopsis. Thus, additional postoperative strategies are needed to promote functional plasticity and enhance the recovery of stereopsis.


Subject(s)
Humans , Exotropia/surgery , Depth Perception/physiology , Strabismus/surgery , Oculomotor Muscles/surgery
15.
Acta Pharmaceutica Sinica B ; (6): 2506-2521, 2022.
Article in English | WPRIM | ID: wpr-929382

ABSTRACT

Retinal pigment epithelial (RPE) is primarily impaired in age-related macular degeneration (AMD), leading to progressive loss of photoreceptors and sometimes choroidal neovascularization (CNV). mTOR has been proposed as a promising therapeutic target, while the usage of its specific inhibitor, rapamycin, was greatly limited. To mediate the mTOR pathway in the retina by a noninvasive approach, we developed novel biomimetic nanocomplexes where rapamycin-loaded nanoparticles were coated with cell membrane derived from macrophages (termed as MRaNPs). Taking advantage of the macrophage-inherited property, intravenous injection of MRaNPs exhibited significantly enhanced accumulation in the CNV lesions, thereby increasing the local concentration of rapamycin. Consequently, MRaNPs effectively downregulated the mTOR pathway and attenuate angiogenesis in the eye. Particularly, MRaNPs also efficiently activated autophagy in the RPE, which was acknowledged to rescue RPE in response to deleterious stimuli. Overall, we design and prepare macrophage-disguised rapamycin nanocarriers and demonstrate the therapeutic advantages of employing biomimetic cell membrane materials for treatment of AMD.

16.
Chinese Journal of Biotechnology ; (12): 4587-4600, 2022.
Article in Chinese | WPRIM | ID: wpr-970333

ABSTRACT

Ultrashort peptides have higher stability, tissue penetrability, biocompatibility, and less immunogenicity, and are widely applied in biology and medicine. GHK (glycyl-l-histidyl-l-lysine) and GQPR (glycyl-l-glutamyl-l-prolyl-l-arginine) can stimulate collagen renewal and inhibit collagen degradation. GHK and GQPR have been used in cosmetic anti-wrinkle skincare and make-up products. The most common approach for ultrashort peptide production is the solid-phase synthesis, which is eco-unfriendly due to heavy usage of organic chemical reagents during the manufacturing process. Here we report a new approach to the production of ultrashort peptides. Recombinant expression of ultrashort peptides is usually unfeasible because of the short amino acid sequences. A vector pET28a-Trxm harboring the thioredoxin gene was first constructed for subsequent fusion expression. The tandem repeats of GHK and GQPR genes were used as the templates for rolling circle amplification (RCA). The RCA reaction was tuned to incorporate noncanonical nucleotides 5-methylcytosine to obtain long DNA fragments. Gene sequences with various lengths were generated through double digestion of Acc65 Ⅰ and Apa Ⅰ. The resulting digestion products were gel recovered by size (from 500 bp to 1 500 bp) and cloned into pET28a-Trxm to obtain the recombinant vector pET28a-Trxm-(TRSP)n. The pET28a-Trxm-(TRSP)n was introduced into E. coli BL21(DE3) to generate a library of Trxm-(TRSP)n sequences with a controlled distribution of lengths. Through double digestion and sequencing, positive clones with tandem repeats n=1, 2, 3, 4, 6, 7, 8, 9 were obtained. Protein expression results showed protein bands with corresponding molecular weight, and the protein expression level decreased as the tandem repeats increased. The expression level of Trxm-(TRSP)1 achieved 50% of the total protein, while the expression level of Trxm-(TRSP)2 was 30% of the total protein. The crude extracts from cell pellets were further treated with enterokinase cleavage, and the supernatants containing (TRSP)1 were collected after ultrafiltration and then subjected to trypsin cleavage. HPLC analysis indicated that the ultrashort peptides GHK and GQPR were successfully obtained through two-step cleavage. This study may facilitate the commercial production of ultrashort peptides.


Subject(s)
Escherichia coli/metabolism , Peptides/chemistry , Amino Acid Sequence , Gene Library , Tandem Repeat Sequences
17.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 337-346, 2022.
Article in Chinese | WPRIM | ID: wpr-940343

ABSTRACT

Through consulting the ancient herbal medicine, prescription books and medical books, combined with modern relevant literature, standards and other information, this paper made a textual research on the name, origin, producing areas, harvesting and processing methods of Astragali Radix according to different historical development periods, providing a basis for the development of famous classical formulas containing Astragali Radix. According to the textual research, the original name of Astragali Radix is Huangqi, and "Qi" originally refers to the medicinal material Zhimu. Some people began to mistake it for Huangqi in the Ming dynasty, and then gradually used Astragali Radix as a medicinal material. The mainstream basis of Astragali Radix can be determined as the dried roots of Astragalus membranaceus var. mongholicus or A. membranaceus. In different historical periods, A. floridus, A. chrysopterus, A. emestii and other plants of Astragalus or even non-Astragalus were used as local Astragali Radix. The earliest production areas of Astragali Radix were Sichuan, Shaanxi, and Gansu, and then gradually expanded to the northeast. Since the Song dynasty, Mianqi in Shanxi province has been regarded as the genuine variety. In the Qing dynasty, besides Shanxi province, Inner Mongolia was also regarded as a genuine place. In the Republic of China, Huangqi produced in northeast China was praised highly. It is mainly produced in Shanxi, Inner Mongolia, Gansu, northeast and other provinces. The main commodity is cultivated products, and the quality of wild imitation cultivation in Datong and Xinzhou is better than other places. There are many processing methods of Huangqi recorded in the materia medica and prescription books, most of which are raw products, and honey processing is the mainstream of processed products. Based on the current situation of resource cultivation and production, 11 famous classical formulas in The Catalogue of Ancient Famous Classical Formulas (The First Batch) containing Huangqi suggested that all use A. membranaceus var. mongholicus, especially those from Datong and Xinzhou in Shanxi Province. In addition to honey processing of Qingxin Lianziyin, it is suggested to use raw products for other formulas.

18.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 327-336, 2022.
Article in Chinese | WPRIM | ID: wpr-940342

ABSTRACT

Through consulting the ancient and modern literature, this paper makes a textual research on the name, origin, producing area, harvesting and processing of Poria, so as to provide a basis for the development of the famous classical formulas containing this medicinal material. The description of Poria and the characteristics of the attached figures in the Chinese herbal literature of the past dynasties are consistent with Poria cocos. The medicinal parts are dried sclerotia or P. cocos peel. Poria was originally produced in Taishan, Shandong province. In the Tang dynasty, along with the change of pine forest resources, producing area of Poria was transferred to Huashan area in Shaanxi province. In the Ming dynasty, the authentic producing area was transferred to Yunnan, and has continued to now. In ancient times, the processing methods of Poria were steaming, boiling, slicing, mashing and other subsequent processing after peeling. It is suggested that Poria in famous classical formulas should be sliced according to the 2020 edition of Chinese Pharmacopoeia.

19.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 318-326, 2022.
Article in Chinese | WPRIM | ID: wpr-940341

ABSTRACT

Through consulting the ancient and modern literature, this paper makes a textual research on the name, origin, producing area, harvesting and processing methods of Asini Corii Colla, so as to provide a basis for the development of the famous classical formulas containing the medicinal material. Before the Tang dynasty, cow leather was the main source of Asini Corii Colla, and donkey was rare as an introduced species. From the end of Tang dynasty to Song dynasty, due to the development of doctors' understanding of the properties and effects of medicines, with the increase of the number of donkeys and the limitation of the use of cow leather, the source of Asini Corii Colla changed from cow leather to donkey skin. During the Ming and Qing dynasties, the theory of medicine property was further developed, and all doctors basically agreed that black donkey skin and E-well water were two essential factors for making genuine Asini Corii Colla. Therefore, it is suggested that Asini Corii Colla should take Equus asinus as the authentic origin in the development of the famous classical formulas, attach importance to the quality of water source, take Liaocheng in Shandong province as the authentic producing area, and the processing should be carried out in accordance with the requirements of the 2020 edition of Chinese Pharmacopoeia.

20.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 306-317, 2022.
Article in Chinese | WPRIM | ID: wpr-940340

ABSTRACT

Based on the ancient literature of all dynasties, this article makes a systematic textual research on the name, origin, producing area, quality, harvesting and processing of Magnoliae Officinalis Cortex used in the famous classical formulas, and clarifies its information of each link in different historical periods, so as to provide a reference and basis for the development and utilization of the related formulas. The results showed that the main varieties of Magnoliae Officinalis Cortex were Magnolia officinalis or M. officinalis var. biloba. The main production areas are Hubei, Sichuan, Chongqing and other places, forming the famous authentic medicine. The processing methods of the past dynasties are mainly cleansing and processing with ginger. In the formulas clearly marked with ginger processing, ginger-processed products is suggested to choose. If not clearly marked, raw or ginger-processed products can be used as needed.

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