2. HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on Dendrobium officinale and Characteristics Analysis of Different Provenances
Zhi-yun LIANG ; Zhen-shan XIE ; Yue-chun HUANG ; Yuan YUAN ; Chu-juan ZHOU ; Ya-wen WANG ; Gang WEI
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(1):22-28
Objective: To optimize the HPLC characteristic spectrum of flavonoid glycosides of Dendrobium officinale,and identify the common and specific components of different provenances. Method: Kromasil 100-5 C18 column was adopted, with tetrahydrofuran-acetonitrile-methanol (10:22:5)-0.05% phosphoric acid as mobile phase (gradient elution). The detection wavelength was 340 nm,the column temperature was 30℃,and the flow rate was 1.0 mL ·min-1. Result: 13 flavonoid characteristic peaks were marked in 27 batches of D. officinale,and 7 characteristic peaks of 6 flavonoid C-glycosides (vicenin Ⅱ,vicenin Ⅰ,schaftoside,isoschaftoside,violanthin and isoviolanthin) and one flavonoid O-glycosides (rutin) was identified. 7-11 characteristic peaks were detected in different batches of samples. Among them,vicenin Ⅱ was a relatively stable common peak in different source samples,and the characteristic peaks of rutin,schaftoside and isoschaftoside were quite different. According to the relative abundance of the characteristic peaks,the samples could be divided into three categories. Among them,the first category had 10 batches of samples,which mainly came from Danxia landforms of Guangdong,Jiangxi,Fujian and Zhejiang (Wuyi) Province (which called "Danxia landform species") and characterized by detection of obvious peak of rutin. The second category had 11 batches of samples,which mainly came from Yunnan and Guangxi Province (which included "Yunnan Guangnan species" and "Guangxi Tiepilan species") and characterized by detection of violanthin and isoviolanthin. And the third category had 6 batches of samples, which were mainly derived from Zhejiang Province (which called "native species from Zhejiang") and characterized by detection of different degrees of rutin peak, but it was difficult to detect the characteristic peaks of violanthin and isoviolanthin. HPLC characteristic chromatograms of D. officinale in bionics wild cultivation and greenhouse of "Danxia landform species" and "Guangxi Tiepilan species" were compared. The results showed that the characteristic peaks in D. officinale planted in greenhouse could be detected stably,which verified the reliability of the source in D. officinale. Conclusion: The analytical method has a better separation effect on flavonoids of D. officinale, with a good reproducibility. The commonness and specificities of flavonoid glycosides components of D. officinale from different categories have basically confirmed. This suggests that Vicenin Ⅱ is suitable to be a reference peak for characteristic chromatogram. Both the relative abundance of rutin and the detection or relative abundance of violanthin and isoviolanthin peaks could be used as a reference to judge the categories of D. officinale in "Danxia landform species" or "Tiepilan species from Yunnan, South Guangdong and Guangxi" or "native species from Zhejiang".
3. Content Comparison of Mannose,Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense
Ya-wen WANG ; Zhi-yun LIANG ; Zhen-shan XIE ; Jia-hong YE ; Si-jian OU ; Chu-juan ZHOU ; Yuan YUAN ; Yue-chun HUANG ; Gang WEI
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(1):35-42
Objective: To optimize the pre-column derivation high performance liquid chromatography (HPLC) content determination method of D-mannose and D-glucose as well as the content determination method of narinhenin in Dendrobium officinale and D. huoshanense, and compare the contents of D-mannose,D-glucose and narinhenin between D. officinale and D. huoshanense. Method: A pre-column derivation HPLC method modified by Chinese Pharmacopoeia(Ch.P) 2015 was used to simultaneously determine the contents of D-mannose and D-glucose,with acetonitrile-0.02 mol·L-1 ammonium acetate solution as mobile phase for gradient elution. Kromasil 100-5 C18 was performed with the wavelength set at 250 nm,and the flow rate was 1 mL·min-1;column temperature was 30℃. HPLC content determination of narinhenin was performed on Kromasil 100-5 C18 with the acetonitrile-methanol-0.4% phosphoric acid solution as mobile phase for gradient elution,and the wavelength was set at 290 nm; the flow rate was 0.8 mL·min-1,and column temperature was 40℃. Result: D-mannose and D-glucose showed a good linear relationship within the range of 0.15-3.0 μg and 0.075-2.25 μg (r=0.999 9); and their average recoveries were 99.01% (RSD 2.1%) and 101.69% (RSD 2.0%) respectively. In addition, the other methodological researches such as repeatability and durability all met the requirements. The contents of D-mannose(Cm),D-glucose(Cg) and sum of them (Cm+Cg) were 12.75%-36.40%,2.93%-18.39% and 19.23%-54.58% in 43 batch of D. officinale. Almost all of the results except very few samples reached the D-mannose standard in Ch.P 2015, and the total content of D-mannose and D-glucose was also up to the total polysccharide standard in Ch.P. The correlation between content and origin was not significant. The contents of D-mannose(Cm),D-glucose(Cg) and sum of them (Cm+Cg) were 14.33%-29.47%,6.64%-15.20%,and 25.73%-44.37% in 12 batch of D. huoshanense. These contents and ratio of peak areas of D-mannose to D-glucose (Am/Ag) were within the scope of D. officinale's; in addition, their average contents were basically the same with those in D. officinale (about 33%).Next,naringenin showed a good linear relationship within the range of 0.020 8-0.832 0 μg (r=0.999 9),and its average recovery was 101.96% (RSD 1.8%). The content of naringenin was 0.053 2-0.122 4 mg·g-1 (average value of 0.081 0 mg·g-1) in 11 batch of D. officinale, slightly higher than 0.040 3-0.090 0 mg ·g-1 (average value of 0.068 3 mg ·g-1) in 7 batch of D. huoshanense. All of these results of narinfenin did not reach the content lower limit in Ch.P. Conclusion: The method used to determinate the content of D-mannose and D-glucose is reproducible, and their sum content is possible to substitute the total polysccaride determination (with higher errors) in D. officinale; monosaccharide content determination can be used for quantitative quality control of D. huoshanense. However, it could not distinguish D. officinale and D. huoshanense by determining the contents of polysccharide,D-glucose,D-mannose and narinhenin, and shall be combined with other specificity methods for further identification.
4.Serosurvey of Avian metapneumovirus, Orithobacterium rhinotracheale, and Chlamydia psittaci and Their Potential Association with Avian Airsacculitis.
Zong Hui ZUO ; Tian Yuan ZHANG ; Yong Xia GUO ; Jun CHU ; Guang Gang QU ; Li Zhong MIAO ; Zhi Qiang SHEN ; Cheng HE
Biomedical and Environmental Sciences 2018;31(5):403-406
Seasonal outbreaks of airsacculitis in China's poultry cause great economic losses annually. This study tried to unveil the potential role of Avian metapneumovirus (AMPV), Ornithobacterium rhinotracheale (ORT) and Chlamydia psittaci (CPS) in avian airsacculitis. A serological investigation of 673 breeder chickens and a case-controlled study of 430 birds were undertaken. Results showed that infection with AMPV, ORT, and CPS was highly associated with the disease. The correlation between AMPV and CPS were positively robust in both layers and broilers. Finally, we determined the co-infection with AMPV, ORT, and CPS was prevalent in the sampled poultry farms suffering from respiratory diseases and the outbreak of airsacculitis was closely related to simultaneous exposure to all three agents.
Air Sacs
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microbiology
;
pathology
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Animals
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Antibodies, Bacterial
;
blood
;
Antibodies, Viral
;
blood
;
Case-Control Studies
;
Chickens
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Chlamydia
;
Chlamydia Infections
;
microbiology
;
pathology
;
veterinary
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Coinfection
;
Flavobacteriaceae Infections
;
microbiology
;
pathology
;
veterinary
;
Humans
;
Metapneumovirus
;
Ornithobacterium
;
Paramyxoviridae Infections
;
pathology
;
veterinary
;
virology
;
Poultry Diseases
;
microbiology
;
pathology
;
virology
;
Respiratory Tract Diseases
;
microbiology
;
veterinary
;
virology
;
Seroepidemiologic Studies
5.Transcriptional and translational responses of rapeseed leaves to red and blue lights at the rosette stage.
Sheng-Xin CHANG ; Chu PU ; Rong-Zhan GUAN ; Min PU ; Zhi-Gang XU
Journal of Zhejiang University. Science. B 2018;19(8):581-595
Under different red (R):blue (B) photon flux ratios, the growth performance of rapeseed (Brassica napus L.) is significantly different. Rapeseed under high R ratios shows shade response, while under high B ratios it shows sun-type morphology. Rapeseed under monochromatic red or blue light is seriously stressed. Transcriptomic and proteomic methods were used to analyze the metabolic pathway change of rapeseed (cv. "Zhongshuang 11") leaves under different R:B photon flux ratios (including 100R:0B%, 75R:25B%, 25R:75B%, and 0R:100B%), based on digital gene expression (DGE) and two-dimensional gel electrophoresis (2-DE). For DGE analysis, 2054 differentially expressed transcripts (|log2(fold change)|≥1, q<0.005) were detected among the treatments. High R ratios (100R:0B% and 75R:25B%) enhanced the expression of cellular structural components, mainly the cell wall and cell membrane. These components participated in plant epidermis development and anatomical structure morphogenesis. This might be related to the shade response induced by red light. High B ratios (25R:75B% and 0R:100B%) promoted the expression of chloroplast-related components, which might be involved in the formation of sun-type chloroplast induced by blue light. For 2-DE analysis, 37 protein spots showed more than a 2-fold difference in expression among the treatments. Monochromatic light (ML; 100R:0B% and 0R:100B%) stimulated accumulation of proteins associated with antioxidation, photosystem II (PSII), DNA and ribosome repairs, while compound light (CL; 75R:25B% and 25R:75B%) accelerated accumulation of proteins associated with carbohydrate, nucleic acid, amino acid, vitamin, and xanthophyll metabolisms. These findings can be useful in understanding the response mechanisms of rapeseed leaves to different R:B photon flux ratios.
Brassica napus/radiation effects*
;
Brassica rapa/radiation effects*
;
Carbon/chemistry*
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Chloroplasts/radiation effects*
;
Computational Biology
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Electrophoresis, Gel, Two-Dimensional
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Gene Expression Regulation, Plant/radiation effects*
;
Image Processing, Computer-Assisted
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Light
;
Mass Spectrometry
;
Metabolic Networks and Pathways
;
Nitrogen/chemistry*
;
Photons
;
Photosystem II Protein Complex/genetics*
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Plant Leaves/radiation effects*
;
Plant Proteins/genetics*
;
Proteome
;
Ribosomes
;
Transcription, Genetic
;
Transcriptome
6.Design space approach to optimize first ethanol precipitation process of Dangshen.
Zhi-lin XU ; Wen-hua HUANG ; Xing-chu GONG ; Tian-tian YE ; Hai-bin QU ; Yan-gang SONG ; Dong-lai HU ; Guo-xiang WANG
China Journal of Chinese Materia Medica 2015;40(22):4411-4416
Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation
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Chemistry, Pharmaceutical
;
methods
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Codonopsis
;
chemistry
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Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
7.Diversity, bioactivities, and metabolic potentials of endophytic actinomycetes isolated from traditional medicinal plants in Sichuan, China.
Peng QIU ; Zhi-Xiang FENG ; Jie-Wei TIAN ; Zu-Chao LEI ; Lei WANG ; Zhi-Gang ZENG ; Yi-Wen CHU ; Yong-Qiang TIAN
Chinese Journal of Natural Medicines (English Ed.) 2015;13(12):942-953
The present study was designed to determine the taxonomic diversity and metabolic activity of the actinomycetes community, including 13 traditional medicinal plants collected in Sichuan province, China, using multiple approaches such as morphological and molecular identification methods, bioactivity assays, and PCR screening for genes involved in antibiotics biosynthesis. 119 endophytic actinomycetes were recovered; 80 representative strains were chosen for 16S rRNA gene partial sequence analyses, with 66 of them being affiliated to genus Streptomyces and the remaining 14 strains being rare actinomycetes. Antimicrobial tests showed that 12 (15%) of the 80 endophytic actinomycetes displayed inhibitory effects against at least one indicator pathogens, which were all assigned to the genus Streptomyces. In addition, 87.5% and 58.8% of the isolates showed anticancer and anti-diabetic activities, respectively. Meanwhile, the anticancer activities of the isolates negatively correlated with their anti-diabetic activities. Based on the results of PCR screening, five genes, PKS-I, PKS-II, NRPS, ANSA, and oxyB, were detected in 55.0%, 58.8%, 90.0%, 18.8% and 8.8% of the 80 actinomycetes, respectively. In conclusion, the PCR screening method employed in the present study was conducive for screening and selection of potential actinomycetes and predicting potential secondary metabolites, which could overcome the limitations of traditional activity screening models.
Actinobacteria
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chemistry
;
classification
;
isolation & purification
;
metabolism
;
Biodiversity
;
China
;
Endophytes
;
chemistry
;
classification
;
isolation & purification
;
metabolism
;
Molecular Sequence Data
;
Phylogeny
;
Plants, Medicinal
;
classification
;
microbiology
;
Secondary Metabolism
8.Unilateral pedicle screw fixation and transforaminal lumbar interbody fusion through paraspinal muscle approach for recurrent lumbar disc herniation combined with lumbar instability.
Bing PAN ; Chu-Di FU ; Yun-Lin GE ; Zhan-You LI ; Zhi-Jing ZHANG ; Hai-Ming LIU ; Gang YANG
China Journal of Orthopaedics and Traumatology 2014;27(9):712-716
OBJECTIVETo explore the clinical effects of unilateral pedicle screw fixation and transforaminal lumbar interbody fusion (TLIF) through paraspinal muscle approach for recurrent lumbar disc herniation combined with lumbar instability.
METHODSA total of 35 patients with recurrent lumbar disc herniation combined with lumbar instability were treated between March 2008 and May 2010, including 15 patients managed by the paraspinal muscle approach with unilateral pedicle screw fixation and transforaminal lumbar interbody fusion (TLIF) (unilateral fixation group) and 20 patients by the posterior midline approach with bilateral pedicle screw fixation and posterior lumbar interbody fusion (bilateral fixation group). Operation time and intraoperative blood loss were observed, preoperative and postoperative JOA score and VAS score in low back pain and legs pain, the interbody fusion condition were compared between two groups.
RESULTSAll patients were followed up from 6 to 30 months with an average 16.8 months. All clinical symptoms had obviously improved postoperatively. X-rays showed good interbody fusion (only 1 case did not obtain fusion in bilateral fixation group) without cage displacement or settlement and implant loosening or breakage. There was significant difference in operation time and the intraoperative blood loss between two groups (P < 0.05). Postoperative JOA score had obviously decreased than preoperative one (P < 0.05). At 1 week after surgery, there was significant difference in VAS score of low back pain between two groups and there was no significant difference in VAS score of legs pain between two groups (P > 0.05); at final follow-up, there was no significant difference in VAS score of low back pain and legs pain between two groups (P > 0.05).
CONCLUSIONTwo methods both can obtain satisfactory effect in treating recurrent lumbar disc herniation combined with lumbar instability. Through the paraspinal muscle approach with unilateral pedicle screw fixation and TLIF has advantages of smaller surgical incision, shorter operation time, less intraoperative blood loss, faster relief in low back pain after operation, etc.
Aged ; Female ; Humans ; Intervertebral Disc Displacement ; physiopathology ; surgery ; Lumbar Vertebrae ; physiopathology ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Recurrence ; Spinal Fusion ; methods
9.Unilateral pedicle screw fixation and transforaminal lumbar interbody fusion through paraspinal muscle approach for recurrent lumbar disc herniation combined with lumbar instability
Bing PAN ; Chu-Di FU ; Yun-Lin GE ; Zhan-You LI ; Zhi-Jing ZHANG ; Hai-Ming LIU ; Gang YANG
China Journal of Orthopaedics and Traumatology 2014;(9):712-716
Objective:To explore the clinical effects of unilateral pedicle screw fixation and transforaminal lumbar inter-body fusion (TLIF) through paraspinal muscle approach for recurrent lumbar disc herniation combined with lumbar instability. Methods:A total of 35 patients with recurrent lumbar disc herniation combined with lumbar instability were treated between March 2008 and May 2010,including 15 patients managed by the paraspinal muscle approach with unilateral pedicle screw fixation and transforaminal lumbar interbody fusion (TLIF) (unilateral fixation group) and 20 patients by the posterior midline approach with bilateral pedicle screw fixation and posterior lumbar interbody fusion (bilateral fixation group). Operation time and intraoperative blood loss were observed ,preoperative and postoperative JOA score and VAS score in low back pain and legs pain,the interbody fusion condition were compared between two groups. Results:All patients were followed up from 6 to 30 months with an average 16.8 months. All clinical symptoms had obviously improved postoperatively. X rays showed good in-terbody fusion (only 1 case did not obtain fusion in bilateral fixation group ) without cage displacement or settlement and im-plant loosening or breakage. There was significant difference in operation time and the intraoperative blood loss between two groups (P<0.05). Postoperative JOA score had obviously decreased than preoperative one (P<0.05). At 1 week after surgery, there was significant difference in VAS score of low back pain between two groups and there was no significant difference in VAS score of legs pain between two groups (P>0.05);at final follow up,there was no significant difference in VAS score of low back pain and legs pain between two groups (P>0.05). Conclusion:Two methods both can obtain satisfactory effect in treating recurrent lumbar disc herniation combined with lumbar instability. Through the paraspinal muscle approach with unilateral pedicle screw fixation and TLIF has advantages of smaller surgical incision,shorter operation time,less intraoperative blood loss,faster relief in low back pain after operation,etc.
10.Preliminary study for classification of spino-pelvic sagittal alignment in adult volunteers.
Gang-Hui YIN ; Ling-Xiang ZHU ; Rui-Song CHEN ; Zhi-de LÜ ; Ming LU ; Hui-Bo YAN ; Zhong-Min ZHANG ; Qing-Chu LI ; Da-di JIN
Chinese Journal of Surgery 2013;51(6):522-526
OBJECTIVETo investigate the feasibility of the classification of the spino-pelvic sagittal alignment in adluts according to lumbar lordosis (LL) and inflection point (IP).
METHODSWhole spine, standing radiographs of 223 adult volunteers were taken from July to August in 2011 .There were 111 cases(56 female and 55 male) enrolled in the study based on the inclusion criteria. The pelvic and spinal parameters, including thoracic kyphosis(TK), thoracolumbar kyphosis(TLK), LL, sacral slope(SS), pelvic tilt(PT), pelvic incidence(PI), intervertebral endplate angle, sagittal vertical axis (SVA), spino-sacral angle (SSA) and IP were measured. The spino-pelvic sagittal alignment were classified in to 3 types according to LL and IP. Type I: LL > -40°, IP located below L2 ∼ 3; Type II: -60° ≤ LL ≤ -40°, IP located in L1 ∼ 2 or T12 ∼ L1; Type III: LL < -60°, P located above T11 ∼ 12. Pearson correlation analysis was used to test the correlation between the variables. The parameters in each type were compared by oneway-ANOVA respectively,then additional multiple comparisons were performed.
RESULTSThe mean value of LL was -49° ± 10°, TK was 36° ± 7°, TLK was 6° ± 7°, PT was 11° ± 7°, SS was 34° ± 8°, PI was 45° ± 9°, SSA was 127° ± 9° and SVA was (-2.7 ± 22.8)mm, respectively. Only LL had significant statistical correlation with all the other parameters. Negative correlation presented between LL and TK, PI, SS, SSA (r = -0.387, -0.536, -0.858, -0.801,P < 0.05). Positive correlation presented between LL and TLK, SVA, PT (r = 0.319, 0.296, 0.262, P < 0.05). All the volunteers were classified into the 3 types: Type I1 9 cases, Type II 75 cases,Type III 17 cases. Oneway-ANOVA results showed statistical difference in LL, TK, TLK, PT, SS, PI, SSA, SVA among the 3 types, (F = 164.559, 7.431, 14.099, 4.217, 53.856, 6.252, 35.995, 8.626, P < 0.05 ). Multiple comparisons showed that LL, SS, SSA, PI had statistical difference between each two types comparison (P < 0.05).
CONCLUSIONSLL is the central parameter of the spino-pelvic sagittal balance. The patterns of the spino-pelvic sagittal alignment in adults could be classified into three types, according to LL and IP. The classification could describe the morphological differences and balance of the spino-pelvic sagittal alignment.
Adult ; Analysis of Variance ; Anthropometry ; Female ; Healthy Volunteers ; Humans ; Male ; Middle Aged ; Pelvis ; anatomy & histology ; Postural Balance ; Radiography ; Spine ; anatomy & histology

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