1.Quality Evaluation of Atractylodis Macrocephalae Rhizoma in Different Production Methods Based on Traditional Traits
Jinxiu QIAN ; Yihan WANG ; Yapeng WANG ; Guoliang YU ; Qiuxiang PAN ; Jiawei SHI ; Meiping CHEN ; Yangqing LIU ; Lun LU ; Yanmeng LIU ; Tiegui NAN ; Liping KANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(1):43-52
ObjectiveBased on the experience of traditional quality evaluation, the quality of Atractylodis Macrocephalae Rhizoma(AMR) with different production methods such as direct seeding, transplanting after seedling raising, topping and non-topping, and difference in growth years was compared. MethodVernier caliper was used to measure the trait data of AMR in different production methods. Paraffin sections of AMR with different production methods were made by saffron solid green staining, and the microstructure was observed. The contents of water-soluble and alcohol-soluble extracts in AMR with different production methods were determined according to the 2020 edition of Chinese Pharmacopoeia. The content of water-soluble total polysaccharides in AMR with different production methods was detected by sulfuric acid-anthrone method. Fiber analyzer was used to detect the content of fiber components in AMR with different production methods. The contents of monosaccharides, oligosaccharides and some secondary metabolites in AMR with different production methods were detected by ultra performance liquid chromatography(UPLC), and the differences of chemical components were compared by multivariate statistical analysis methods such as principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA). ResultIn terms of traits, the 3-year-old AMR with direct seeding and without topping was close to the high-quality AMR with "phoenix-head and crane-neck, strong sweetness and clear aroma" recorded in ancient materia medica, followed by the 3-year-old AMR with topping after transplanting, while the 2-year-old AMR with topping after transplanting with high market circulation rate was generally fat and strong with mild odor. In the microscopic aspect, the arrangement of xylem vessels and fiber bundles in the 3-year-old samples formed two obvious rings. Compared with the 2-year-old samples cultivated in Bozhou and Zhejiang, the 3-year-old samples without topping after transplanting had more wood fibers. In terms of chemical composition, the contents of 70% ethanol extract, fructose, glucose, sucrose, 1-kestose, atractylenolide Ⅰ, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid and other components in 3-year-old AMR with direct seeding and without topping were significantly higher than those in the other three samples(P<0.05). The contents of cellulose, 70% ethanol extract, sucrose, atractylenolide Ⅰ, atractylone and other components in 3-year-old AMR with topping after transplanting were significantly higher than those in the 2-year-old AMR with high market circulation rate(P<0.05), while the contents of water-soluble extract and water-soluble total polysaccharides in 2-year-old samples with topping after transplanting were significantly higher than those in the 3-year-old AMR with topping after transplanting, direct seeding and without topping(P<0.05). ConclusionUnder the current mainstream production mode, too much manual intervention makes AMR heavily enriched in polysaccharides and increased the yield, but the accumulation of sweet substances, fragrant substances and fiber substances is insufficient, which affects its quality. The current quality standard of AMR has some shortcomings in guiding the high quality production of it, it is suggested to revise the quality standard of AMR, supplement the quantitative analysis of secondary metabolites, and strengthen the production of imitation wild AMR.
2.Late-stage cascade of oxidation reactions during the biosynthesis of oxalicine B in Penicillium oxalicum.
Tao ZHANG ; Guowei GU ; Guodong LIU ; Jinhua SU ; Zhilai ZHAN ; Jianyuan ZHAO ; Jinxiu QIAN ; Guowei CAI ; Shan CEN ; Dewu ZHANG ; Liyan YU
Acta Pharmaceutica Sinica B 2023;13(1):256-270
Oxalicine B ( 1) is an α-pyrone meroterpenoid with a unique bispirocyclic ring system derived from Penicillium oxalicum. The biosynthetic pathway of 15-deoxyoxalicine B ( 4) was preliminarily reported in Penicillium canescens, however, the genetic base and biochemical characterization of tailoring reactions for oxalicine B ( 1) has remained enigmatic. In this study, we characterized three oxygenases from the metabolic pathway of oxalicine B ( 1), including a cytochrome P450 hydroxylase OxaL, a hydroxylating Fe(II)/α-KG-dependent dioxygenase OxaK, and a multifunctional cytochrome P450 OxaB. Intriguingly, OxaK can catalyze various multicyclic intermediates or shunt products of oxalicines with impressive substrate promiscuity. OxaB was further proven via biochemical assays to have the ability to convert 15-hydroxdecaturin A ( 3) to 1 with a spiro-lactone core skeleton through oxidative rearrangement. We also solved the mystery of OxaL that controls C-15 hydroxylation. Chemical investigation of the wild-type strain and deletants enabled us to identify 10 metabolites including three new compounds, and the isolated compounds displayed potent anti-influenza A virus bioactivities exhibiting IC50 values in the range of 4.0-19.9 μmol/L. Our studies have allowed us to propose a late-stage biosynthetic pathway for oxalicine B ( 1) and create downstream derivatizations of oxalicines by employing enzymatic strategies.
3.Research advances on invasive fungal infections after burns
Yue CAI ; Xing CHENG ; Jianhua ZHAN ; Jinhua LUO ; Wenqiang LIAO
Chinese Journal of Burns 2023;39(3):269-274
Invasive fungal infection (IFI) is one of the serious complications in burn patients. The gradual development and application of broad-spectrum antibiotics in recent years has led to a serious dysbiosis of the flora, while the widespread prophylactic use of antifungal drugs has led to an increasing number of drug-resistant fungi. The clinical treatment of IFI is difficult and the prognosis is poor. The mortality of burn patients caused by IFI is increasing year by year. This paper reviews the epidemiologic characteristics, related risk factors, diagnostic methods, and treatment progress of IFI after burns, aiming to provide new ideas and reference for the prevention and treatment of IFI after burns.
4.Effect of alprostadil in early wound healing of rats with deep partial-thickness scald
Xing CHENG ; Jianhua ZHAN ; Jinhua LUO ; Guoyu ZHENG
Chinese Journal of Burns 2020;36(4):317-322
Objective:To explore the effect of alprostadil in early wound healing of rats with deep partial-thickness scald.Methods:Ninety specific pathogen free healthy Sprague-Dawley rats (half males and half females) were divided into sham scald group, simple scald group, and alprostadil group using the random number table with 30 rats in each group. Rats in sham scald group were sham injured, and rats in the other 2 groups were inflicted with deep partial-thickness scald of 30% total body surface area on the back. Immediately after scald, rats in the 3 groups received anti-shock treatment. Two hours post scald, rats in sham scald group and simple scald group were injected intraperitoneally with 1 mL normal saline, and rats in alprostadil group were injected intraperitoneally with 1 mL alprostadil injection, once a day and continued for 14 days. On post scald day (PSD) 3, 7, and 14, 10 rats in each group were collected for observing the general wound conditions and the wound healing rates of rats in 2 scald groups were calculated, abdominal aortic blood of 2 mL from each rat were collected to detect serum thromboxane B2 by enzyme-linked immunosorbent assay, and wound tissue on the back was collected to examine pathomorphological change by hematoxylin-eosin staining and to detect wound microvessel density (MVD) by immunohistochemical staining. Data were statistically analyzed with analysis of variance of factorial design, t test, Kruskal-Wallis H test, Mann-Whitney U test, and Bonferroni correction. Results:(1) There was no scald wound in rats in sham scald group. On PSD 3, wounds of rats in simple scald group and alprostadil group formed dry eschar. On PSD 7 and 14, wound areas of rats in alprostadil group were significantly smaller than those of rats in simple scald group, and with less exudation. (2) On PSD 3, the wound healing rates of rats in simple scald group and alprostadil group were similar ( t=1.167, P>0.05). On PSD 7 and 14, the wound healing rates of rats in alprostadil group were significantly higher than those in simple scald group ( t=8.657, 33.050, P<0.01). (3) On PSD 3, 7, and 14, the levels of serum thromboxane B2 of rats in simple scald group and alprostadil group were (541±22), (607±47), (688±21), (326±25), (271±21), (135±27) pg/mL, significantly higher than (17±6), (16±4), (16±4) pg/mL of rats in sham scald group ( t=72.977, 39.685, 102.076, 37.033, 37.253, 13.845, P<0.01). On PSD 3, 7, and 14, the levels of serum thromboxane B2 of rats in alprostadil group were significantly lower than those in simple scald group ( t=20.637, 20.651, 51.680, P<0.01). (4) Normal epidermis and dermis were seen in rats in sham scald group. On PSD 3, a large number of necrotic tissue and inflammatory cells infiltration were seen in wounds of rats in simple scald group, while a little new epithelium formation and some inflammatory cells infiltration were seen in wounds of rats in alprostadil group. On PSD 7 and 14, the new epithelium of rats in alprostadil group was significantly thicker than that in simple scald group, and epidermis formed gradually in alprostadil group. (5) On PSD 3, 7, and 14, the wound MVD of rats in simple scald group and alprostadil group were significantly higher than those in sham scald group ( Z=-3.780, -3.781, -3.780, -3.780, -3.781, -3.780, P<0.01). On PSD 3, the wound MVD of rats in simple scald group and alprostadil group were similar ( Z=-1.965, P>0.05). On PSD 7 and 14, the wound MVD of rats in alprostadil group were significantly higher than those in simple scald group ( Z=-3.780, -3.780, P<0.01). Conclusions:The early intervention with alprostadil can significantly improve microcirculation of deep partial-thickness scald wound, reduce inflammatory cell infiltration, promote the formation of new blood vessels, thus promoting wound healing.
5.Tandem expression and activity determination of antibacterial peptide Spinosan-C from Paa spinosa.
Yue LIU ; Zhonggen ZHAN ; Bing ZHU ; Rongquan ZHENG ; Hongyi CHENG ; Zuoming NIE
Chinese Journal of Biotechnology 2018;34(1):132-139
Antibacterial peptide can be easily degraded by protease and has the lethal effect on the host Escherichia coli. In order to solve these problems and further improve the expression ability of the Escherichia coli system, the antimicrobial peptide Spinosan-C of Paa spinosa was studied. First, the codon of Spinosan-C was optimized according to E. coli codon usage frequency. Then, the 8 multimeric Spinosan-C gene (8×Spinosan-C) was synthesized and cloned into prokaryotic expression vector pET-28a. The fusion antimicrobial peptide 8×Spinosan-C was further highly expressed in Escherichia coli strain Rosetta. The recombinant 8×Spinosan-C protein was then purified and cleaved specially by formic acid to generate the Spinosan-C monomer. Antibacterial test in vitro suggested that the cleaved Spinosan-C monomer had antibacterial bioactivity against the test bacteria. This study provides a technical reference for the largescale preparation of frog antimicrobial peptides.
6. Effect of alprostadil on wound healing of scalded rats and the mechanism
Guoyu ZHENG ; Jianhua ZHAN ; Jinhua LUO ; Xing CHENG
Chinese Journal of Burns 2018;34(6):380-385
Objective:
To explore effect of alprostadil on wound healing of scalded rats and the mechanism.
Methods:
According to random number table method, forty-eight Sprague Dawley rats were divided into sham scald group, simple scald group, lithium chloride group, and alprostadil group, with 12 rats in each group. Rats in sham injury group were sham injured on the back, and rats in the other three groups were inflicted with 30% total body surface area deep partial thickness scald on the back.Immediately after scald, rats in sham scald group and simple scald group were injected with 1 mL saline through caudal vein, and rats in lithium chloride group and alprostadil group were injected respectively with 1 mL lithium chloride and alprostadil through caudal vein. Saline, lithium chloride, and alprostadil were injected once in a day and lasted for 14 days. General wound appearance and wound healing rate on post scald day (PSD) 7, 10, 14 were observed and calculated. Expressions of protein and mRNA of Wnt1 and β-catenin on PSD 14 were detected. Data were processed with analysis of variance of factorial design, one-way analysis of variance, Student Newman Keuls
7. Effects of astragalus polysaccharide on cardiac dysfunction in rabbits with severe scald injury
Lin ZHONG ; Jianhua ZHAN ; Jinhua LUO ; Xing CHENG
Chinese Journal of Burns 2017;33(11):668-676
Objective:
To investigate the effects of astragalus polysaccharide (AP) on cardiac dysfunction in rabbits with severe scald injury.
Methods:
Sixty-four New Zealand white rabbits were divided into pure scald group and AP group according to the random number table, with 32 rabbits in each group. Rabbits in the two groups were all inflicted with 30% total body surface area full-thickness scald on the back. Immediately after injury, rabbits in two groups were intraperitoneally injected with lactated Ringer′s solution once for antishock. Rabbits in AP group were intraperitoneally injected with 10 mL AP solution with the dosage of 200 mg/kg 10 min after injury and the following 6 days respectively, once a day. Rabbits in pure scald group were injected with 10 mL normal saline instead. Eight rabbits of each group were respectively selected before injury hour (BIH) 1 and on post injury day(PID) 1, 3, 7, and 14 to collect blood samples from ear marginal vein, and then sacrificed immediately to collect hearts at each time point post injury. The morphology of myocardium was observed after HE staining. The serum content of cardiac troponin I (cTnI) was detected by enzyme-linked immunosorbent assay (ELISA). The serum content of aspartate transaminase (AST), creatine kinase (CK), CK isoenzyme-MB (CK-MB), lactate dehydrogenase (LDH) was detected by fully automatic chemistry analyzer. The content of angiotensin Ⅱ (Ang Ⅱ) in serum and myocardium was detected with radioimmunoassay and the content of endothelin 1 (ET-1) in serum and myocardium was detected by ELISA. Another 8 normal rabbits were sacrificed to detect the content of Ang Ⅱ and ET-1 in the myocardium as the value of the two groups of scalded rabbits at BIH 1. The serum content of superoxide dismutase (SOD) and malondialdehyde (MDA) was detected by ELISA. The values of whole blood viscosity (ηb), reductive viscosity of whole blood (ηr), plasma viscosity (ηp), hematocrit (HCT), erythrocyte rigidity index (TK), erythrocyte aggregation index (EAI), and erythrocyte sedimentation rate (ESR) were detected by fully automatic hematology analyzer. Data were processed with analysis of variance of factorial design, independent sample
8.Effects of astragalus polysaccharide on intestinal immune function of rats with severe scald injury.
Cuilan HUANG ; Jianhua ZHAN ; Jinhua LUO
Chinese Journal of Burns 2015;31(1):30-36
OBJECTIVETo observe the effects of astragalus polysaccharide (AP) on the intestinal mucosal morphology, level of secretory IgA (s-IgA) in intestinal mucus, and distribution of T lymphocyte subsets in Peyer's patch in rats with severe scald injury.
METHODSOne hundred and thirty SD rats were divided into sham injury group (SI, sham injured, n = 10), scald group (S, n = 30), low dosage group (LD, n = 30), moderate dosage group (MD, n = 30), and high dosage group (HD, n = 30) according to the random number table. Rats in the latter 4 groups were inflicted with 30% TBSA full-thickness scald on the back. From post injury hour 2, rats in groups LD, MD, and HD were intraperitoneally injected with 0.5 mL AP solution with the dosage of 100, 200, and 300 mg/kg each day respectively, and rats in group S were injected with 0.5 mL normal saline instead. Ten rats from group SI immediately after injury and 10 rats from each of the latter 4 groups on post injury day (PID) 3, 7, 14 were sacrificed, and their intestines were harvested. The morphology of ileal mucosa was examined after HE staining; the level of s-IgA in ileal mucus was determined with double-antibody sandwich ELISA method; the proportions of CD3⁺, CD4⁺, CD8⁺ T lymphocytes in Peyer's patches of intestine were determined with flow cytometer, and the proportion of CD4⁺ to CD8⁺ was calculated. Data were processed with one-way analysis of variance, analysis of variance of factorial design, and SNK test.
RESULTS(1) Villi in normal form and intact villus epithelial cells were observed in rats of group SI immediately after injury, while edema of villi and necrosis and desquamation of an enormous amount of villi were observed in groups with scalded rats on PID 3, with significant infiltration of inflammatory cells. On PID 7, no obvious improvement in intestinal mucosal lesion was observed in groups with scalded rats. On PID 14, the pathology in intestinal mucosa of rats remained nearly the same in group S, and it was alleviated obviously in groups LD and MD, and the morphology of intestinal mucosa of rats in group HD was recovered to that of group SI. (2) On PID 3, 7, and 14, the level of s-IgA in intestinal mucus significantly decreased in groups S, LD, MD, and HD [(43 ± 5), (45 ± 5), (46 ± 5) µg/mL; (47 ± 5), (48 ± 5), (49 ± 6) µg/mL; (50 ± 6), (51 ± 5), (52 ± 5) µg/mL; (53 ± 6), (54 ± 5), (55 ± 5) µg/mL] as compared with that of rats in group SI immediately after injury [(69 ± 4) µg/mL, with P values below 0.05]. The level of s-IgA in intestinal mucus of rats in group MD was significantly higher than that in group S at each time point (with P values below 0.05), and that of group HD was significantly higher than that in groups S and LD at each time point (with P values below 0.05). (3) Compared with those of rats in group SI immediately after injury, the proportions of CD3⁺ T lymphocytes and CD4⁺ T lymphocytes significantly decreased in groups with scalded rats at each time point (with P values below 0.05), except for those in group HD on PID 14. The proportion of CD4⁺ T lymphocytes of rats in group LD was significantly higher than that in group S on PID 3 (P < 0.05). The proportions of CD3⁺ T lymphocytes and CD4⁺ T lymphocytes were significantly higher in groups MD and HD than in groups S and LD (except for the proportion of CD4⁺ T lymphocytes in group MD on PID 3 and 14) at each time point (with P values below 0.05). The proportion of CD3⁺ T lymphocytes on PID 7 and 14 and that of CD4⁺ T lymphocytes on PID 3 were significantly higher in group HD than in group MD (with P values below 0.05). Compared with that of rats in group SI immediately after injury, the proportion of CD8⁺ T lymphocytes significantly increased in the other 4 groups at each time point (with P values below 0.05). The proportion of CD8⁺ T lymphocytes was significantly lower in rats of group LD on PID 7 and 14 and groups MD and HD at each time point than in group S (with P values below 0.05). The proportion of CD8⁺ T lymphocytes was significantly lower in rats of group MD on PID 7 and 14 and group HD at each time point than in group LD (with P values below 0.05). The proportion of CD8⁺ T lymphocytes was significantly lower in rats of group HD on PID 7 and 14 than in group MD (with P values below 0.05). On PID 3, 7, and 14, the proportion of CD4⁺ to CD8⁺ was significantly lower in groups S, LD, MD, and HD (0.65 ± 0.11, 0.68 ± 0.13, 0.73 ± 0.22; 0.76 ± 0.15, 0.78 ± 0.14, 0.90 ± 0.10; 0.85 ± 0.21, 0.89 ± 0.18, 1.08 ± 0.19; 0.99 ± 0.20, 1.05 ± 0.21, 1.25 ± 0.23) as compared with that of rats in group SI immediately after injury (1.74 ± 0.20, with P values below 0.05). The proportion of CD4⁺ to CD8⁺ was significantly higher in rats of group HD than in group MD on PID 7 (P < 0.05), and the proportion was significantly higher in these two groups than in group S at each time point (with P values below 0.05). The proportion of CD4⁺ to CD8⁺ was significantly higher in rats of group MD on PID 14 and group HD at each time point than in group LD (with P values below 0.05). Compared within each group, the proportions of CD3⁺, CD4⁺, CD8⁺ T lymphocytes and the proportion of CD4⁺ to CD8⁺ of rats in groups LD, MD, and HD showed a trend of gradual elevation along with passage of time.
CONCLUSIONSAP can improve the injury to intestinal mucosa and modulate the balance of T lymphocyte subsets in Peyer's patch in a time- and dose-dependent manner, and it can promote s-IgA secretion of intestinal mucosa in a dose-dependent manner.
Animals ; Astragalus Plant ; adverse effects ; Burns ; immunology ; pathology ; physiopathology ; Dose-Response Relationship, Drug ; Immunity, Mucosal ; Immunoglobulin A ; metabolism ; Intestinal Mucosa ; metabolism ; physiology ; Intestine, Small ; metabolism ; Peyer's Patches ; immunology ; physiopathology ; Polysaccharides ; Rats ; Rats, Sprague-Dawley ; Soft Tissue Injuries ; T-Lymphocyte Subsets ; immunology
9.Key points of digestive tract anastomosis in pancreatic surgery assisted by da Vinci robotic surgical system
Baiyong SHEN ; Jinhua YE ; Qian ZHAN
Chinese Journal of Digestive Surgery 2013;(5):328-331
The da Vinci robotic surgical system has the advantages of three-dimensional vision and high degree of accuracy,flexibility and repeatability,which makes surgical procedures such as digestive tract anastomosis easier to conduct under minimally invasive conditions.In this article,the feasibility and principle of digestive tract anastomosis and the procedures of pancreaticojejunostomy and pancreaticogastrostomy by the da Vinci robotic surgical system are introduced,so as to improve the quality of anastomosis and reduce the incidences of postoperative complications.Compared with traditional laparotomy,da Vinci robotic surgical system simplified the surgical procedures and reduced the trauma,which is suitable for digestive tract anastomosis in pancreatic surgery.The method of pancreatic anastomosis should be selected in consideration of the condition of patients,surgical procedure and the experience of surgeons.
10.Preparation of anti-salbutamol antibody based on a new designed immunogen and development of a heterologous indirect ELISA for detection of salbutamol residue.
Meng MENG ; Yulan ZHANG ; Shengxin LU ; Jinting LIU ; Jinhua ZHAN ; Rimo XI
Acta Pharmaceutica Sinica 2010;45(4):442-50
To synthesize salbutamol immunogen and develop an enzyme immunoassay (ELISA), a new salbutamol immunogen was synthesized using 4-aminobenzoic acid as a linker to connect hapten with carrier protein. An enzyme immunoassay based on the antibody prepared was developed and applied to detect salbutamol residue spiked in swine liver. An unusual coating antigen, clenbuterol-ovalbumin (OVA) conjugate instead of salbutamol-OVA conjugate, was used in the immunoassay and the results were discussed based on the structures of related compounds. The antibodies showed high sensitivity in the heterologous assay when using clenbuterol-OVA as a coating antigen, with an IC50 value of 8.97 ng mL(-1) toward salbutamol. The antibodies prepared showed high cross-reactivity with clenbuterol (107%) and were promising for the simultaneous determination of salbutamol and clenbuterol residues in food and food products. Recovery rates from the salbutamol-spiked swine liver samples were in the range of 70%-99%, while the intra-assay and inter-assay coefficients of variation were <13.3% and <14.3%, respectively. In summary, the antibodies of salbutamol have been successfully prepared. Sensitive and stable analysis for the detection of salbutamol residues in swine liver was obtained based on the competitive ELISA methods developed in this study.

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