Publications by authors named "Zhenjie Cao"

24 Publications

  • Page 1 of 1

Do Various Treatment Modalities of Vesicoureteral Reflux Have Any Adverse Effects in Pediatric Patients? A Meta-Analysis.

Urol Int 2021 Sep 23:1-9. Epub 2021 Sep 23.

Department of Pediatrics, Peking University First Hospital, Beijing, China.

Purpose: Vesicoureteral reflux (VUR) is a risk factor for various renal problems like recurrent urinary tract infections (UTIs), pyelonephritis, renal scarring, hypertension, and other renal parenchymal defects. The interventions followed by pediatricians include low-dose antibiotic treatment, surgical correction, and endoscopy. This meta-analysis aimed to assess the advantages and drawbacks of various primary VUR treatment options.

Search Strategy: The Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, reference lists of journals, and abstracts from conference proceedings were all used to find randomized controlled trials. The articles were retrieved from 1985 till 2020. Twenty articles were used for the data analysis. Criteria for Selection: Surgery, long-term antibiotic prophylaxis, noninvasive techniques, and any mix of therapies are also options for treating VUR. Collection and Interpretation of Data: Two authors searched the literature separately, determining research qualifications, assessing accuracy, and extracting and entering results. The odds ratio (OR) of these studies was used to construct the forest plot. The random-effects model was used to pool the data. Also, the random-effects model was used with statistical significance at a p value < 0.05 to assess the difference in side effects after treatment of VUR using different modalities.

Results: We found no statistically significant differences between surgery plus antibiotics and antibiotic alone-treated patients in terms of recurrent UTIs (OR = 0.581; 95% confidence interval [CI] 0.259-1.30), renal parenchymal defects (OR = 1.149; 95% CI 0.75-1.754), and renal scarring (OR = 1.042; 95% CI 0.72-1.50). However, the risk of developing pyelonephritis after surgical treatment of VUR was lesser than that in the conservative approach, that is, antibiotics (OR = 0.345; 95% CI 0.126-0.946.), positive urine culture (OR = 0.617; 95% CI 0.428-0.890), and recurrent UTIs were more common in the placebo group than in the antibiotic group (p < 0.05; OR = 0.639; 95% CI 0.436-0.936) which is statistically significant.

Conclusion: Based on current research, we recommend that a child with a UTI and significant VUR be treated conservatively at first, with surgical care reserved for children who have issues with antimicrobials or have clinically significant VUR that persists after several years of follow-up.
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http://dx.doi.org/10.1159/000518603DOI Listing
September 2021

The added value of an artificial intelligence system in assisting radiologists on indeterminate BI-RADS 0 mammograms.

Eur Radiol 2021 Sep 15. Epub 2021 Sep 15.

Department of Radiology, Shenzhen People's Hospital, Shenzhen, 518020, Guangdong, China.

Objectives: To investigate the value of an artificial intelligence (AI) system in assisting radiologists to improve the assessment accuracy of BI-RADS 0 cases in mammograms.

Methods: We included 34,654 consecutive digital mammography studies, collected between January 2011 and January 2019, among which, 1088 cases from 1010 unique patients with initial BI-RADS 0 assessment who were recalled during 2 years of follow-up were used in this study. Two mid-level radiologists retrospectively re-assessed these BI-RADS 0 cases with the assistance of an AI system developed by us previously. In addition, four entry-level radiologists were split into two groups to cross-read 80 cases with and without the AI. Diagnostic performance was evaluated using the follow-up diagnosis or biopsy results as the reference standard.

Results: Of the 1088 cases, 626 were actually normal (BI-RADS 1 and no recall required). Assisted by the AI system, 351 (56%) and 362 (58%) normal cases were correctly identified by the two mid-level radiologists hence can be avoided for unnecessary follow-ups. However, they would have missed 12 (10 invasive cancers and 2 ductal carcinoma in situ cancers) and 6 (invasive cancers) malignant lesions respectively as a result. These missed lesions were not highly malignant tumors. The inter-rater reliability of entry-level radiologists increased from 0.20 to 0.30 (p < 0.005) by introducing the AI.

Conclusion: The AI system can effectively assist mid-level radiologists in reducing unnecessary follow-ups of mammographically indeterminate breast lesions and reducing the benign biopsy rate without missing highly malignant tumors.

Key Points: • The artificial intelligence system could assist mid-level radiologists in effectively reducing unnecessary BI-RADS 0 mammogram recalls and the benign biopsy rate without missing highly malignant tumors. • The artificial intelligence system was capable of detecting low suspicion lesions from heterogeneously and extremely dense breasts that radiologists tended to miss. • The use of an artificial intelligence system may improve the inter-rater reliability and sensitivity, and reduce the reading time of entry-level radiologists in assessing potential lesions in BI-RADS 0 mammograms.
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http://dx.doi.org/10.1007/s00330-021-08275-0DOI Listing
September 2021

Establishment of a new fish cell line from the brain of humpback grouper (Cromileptes altivelis) and its application in toxicology and bacterial susceptibility.

Fish Physiol Biochem 2021 Oct 26;47(5):1645-1658. Epub 2021 Aug 26.

Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province, Haikou, Hainan, 570228, People's Republic of China.

Cromileptes altivelis, humpback grouper, belongs to the family Epinephelidae and is one popular farmed fish species because of its high economic value and ornamental value. However, more and more diseases outbreaks have been reported with C. altivelis aquaculture. Today, a new brain cell line of C. altivelis (named CAB) was established and characterized. Our results showed that CAB cells were suitable for growth at 26 °C in L-15 medium supplemented with 15% fetal bovine serum (FBS). The results of 18S rRNA gene sequencing confirmed that CAB cell line was derived from C. altivelis. Moreover, chromosomal aneuploidy was observed in CAB cells, and the modal chromosome number of CAB cells was 48 by chromosome analysis. In addition, CAB cells could transfect pEGFP-N3 plasmid with high transfection efficiency, indicating that CAB cell line has the potential to investigate the function of exogenous genes in vitro. Furthermore, the bacterial susceptibility results suggested that CAB cells were susceptive to Vibrio harveyi and Edwardsiella tarda. And, heavy metals (Hg, Cd, and Cu) were toxic to the CAB cells, and the toxic effect was dose-dependent. In summary, the CAB cell line could be a powerful tool in vitro to study functional genes and has the potential application in bacterial susceptibility and toxicology.
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http://dx.doi.org/10.1007/s10695-021-01006-8DOI Listing
October 2021

MommiNet-v2: Mammographic multi-view mass identification networks.

Med Image Anal 2021 10 2;73:102204. Epub 2021 Aug 2.

Department of Radiology, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University), Shenzhen, Guangdong 518020, China.

Many existing approaches for mammogram analysis are based on single view. Some recent DNN-based multi-view approaches can perform either bilateral or ipsilateral analysis, while in practice, radiologists use both to achieve the best clinical outcome. MommiNet is the first DNN-based tri-view mass identification approach, which can simultaneously perform bilateral and ipsilateral analysis of mammographic images, and in turn, can fully emulate the radiologists' reading practice. In this paper, we present MommiNet-v2, with improved network architecture and performance. Novel high-resolution network (HRNet)-based architectures are proposed to learn the symmetry and geometry constraints, to fully aggregate the information from all views for accurate mass detection. A multi-task learning scheme is adopted to incorporate both Breast Imaging-Reporting and Data System (BI-RADS) and biopsy information to train a mass malignancy classification network. Extensive experiments have been conducted on the public DDSM (Digital Database for Screening Mammography) dataset and our in-house dataset, and state-of-the-art results have been achieved in terms of mass detection accuracy. Satisfactory mass malignancy classification result has also been obtained on our in-house dataset.
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http://dx.doi.org/10.1016/j.media.2021.102204DOI Listing
October 2021

Health-adjusted life expectancy (HALE) in Chongqing, China, 2017: An artificial intelligence and big data method estimating the burden of disease at city level.

Lancet Reg Health West Pac 2021 Apr 2;9:100110. Epub 2021 Mar 2.

Ping An Technology (Shenzhen) Co., Ltd., Ping'an International Financial Center, Futian District, Shenzhen 518001, China.

Background: A universally applicable approach that provides standard HALE measurements for different regions has yet to be developed because of the difficulties of health information collection. In this study, we developed a natural language processing (NLP) based HALE estimation approach by using individual-level electronic medical records (EMRs), which made it possible to calculate HALE timely in different temporal or spatial granularities.

Methods: We performed diagnostic concept extraction and normalisation on 13•99 million EMRs with NLP to estimate the prevalence of 254 diseases in WHO Global Burden of Disease Study (GBD). Then, we calculated HALE in Chongqing, 2017, by using the life table technique and Sullivan's method, and analysed the contribution of diseases to the expected years "lost" due to disability (DLE).

Findings: Our method identified a life expectancy at birth (LE) of 77•9 years and health-adjusted life expectancy at birth (HALE) of 71•7 years for the general Chongqing population of 2017. In particular, the male LE and HALE were 76•3 years and 68•9 years, respectively, while the female LE and HALE were 80•0 years and 74•4 years, respectively. Cerebrovascular diseases, cancers, and injuries were the top three deterioration factors, which reduced HALE by 2•67, 2•15, and 1•19 years, respectively.

Interpretation: The results demonstrated the feasibility and effectiveness of EMRs-based HALE estimation. Moreover, the method allowed for a potentially transferable framework that facilitated a more convenient comparison of cross-sectional and longitudinal studies on HALE between regions. In summary, this study provided insightful solutions to the global ageing and health problems that the world is facing.

Funding: National Key R and D Program of China (2018YFC2000400).
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http://dx.doi.org/10.1016/j.lanwpc.2021.100110DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8315391PMC
April 2021

Functional characterization of cathepsin B and its role in the antimicrobial immune responses in golden pompano (Trachinotus ovatus).

Dev Comp Immunol 2021 Oct 31;123:104128. Epub 2021 May 31.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China.

Cathepsin B (CTSB) is one of the typical representatives of cysteine protease family. It has the activity of both exopeptidase and endopeptidase. It plays an important role in antigen presentation, degradation, apoptosis, inflammatory response and physiological process of many diseases. In this study, CTSB of Trachinotus ovatus (TroCTSB) was cloned, and its structure and function were analyzed. The results showed that the coding region of TroCTSB was 993 bp, encoding 330 amino acid residues. The homology analysis showed that the amino acid sequence of TroCTSB was similar to that in other teleosts and mammals (68.69%-88.48%). Under normal physiological conditions, TroCTSB was widely distributed in various tissues with the highest expression level in stomach, followed by liver, and the lowest expression level in blood. The optimal pH and temperature of purified recombinant protein rTroCTSB were 5.5 and 40 °C, respectively. The toxicity test of metal ions showed that Fe, Cu, Ca and Zn could all inhibit the activity of TroCTSB, with Zn ranking the first. In addition, after Edwardsiella tarda infection, the expression of TroCTSB was significantly up-regulated in liver, spleen and head kidney. The overexpression of TroCTSB significantly inhibited the infection of E. tarda in golden pompano tissues, and the knockdown of TroCTSB remarkably promoted the reproduction of E. tarda in golden pompano tissues in vivo. This study suggests that TroCTSB was involved in the antibacterial immune response of T. ovatus, and provided a reference for further research in elucidating the resistance mechanism of TroCTSB.
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http://dx.doi.org/10.1016/j.dci.2021.104128DOI Listing
October 2021

R848 Is Involved in the Antibacterial Immune Response of Golden Pompano () Through TLR7/8-MyD88-NF-κB-Signaling Pathway.

Front Immunol 2020 18;11:617522. Epub 2021 Jan 18.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, China.

R848 is an imidazoquinoline compound that is a specific activator of toll-like receptor (TLR) 7/8 and is often used in immunological research in mammals and teleosts. However, the immune responses initiated by R848 through the TLR7/8 pathway in response to bacterial infection remain largely unexplored in teleosts. In the current study, we investigated the antibacterial response and the participating signaling pathway initiated by R848 in golden pompano (). We found that R848 could stimulate the proliferation of head kidney lymphocytes (HKLs) in a dose-dependent manner, enhance the survival rate of HKLs, and inhibit the replication of bacteria . However, these effects induced by R848 were significantly reduced when chloroquine (CQ) was used to blocked endosomal acidification. Additionally, an study showed that R848 strengthened the antibacterial immunity of fish through a TLR7/8 and Myd88-dependent signaling pathway. A cellular experiment showed that Pepinh-MYD (a Myd88 inhibitor) significantly reduced the R848mediated proliferation and survival of HKLs. Luciferase activity analysis showed that R848 enhanced the nuclear factor kappa B (NF-κB) activity, whereas this activity was reduced when CQ and Pepinh-MYD were present. Additionally, when an NF-κB inhibitor was present, the R848-mediated pro-proliferative and pro-survival effects on HKLs were significantly diminished. An study showed that knockdown of TLR7, TLR8, and Myd88 expression in golden pompano siRNA following injection of R848 resulted in increased bacterial dissemination and colonization in fish tissues compared to that of fish injection of R848 alone, suggesting that R848-induced antibacterial immunity was significantly reduced. In conclusion, these results indicate that R848 plays an essential role in the antibacterial immunity of golden pompano the TLR7/8-Myd88-NF-κB- signaling pathway.
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http://dx.doi.org/10.3389/fimmu.2020.617522DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7848160PMC
June 2021

Interferon regulatory factor 7 contributes to the host response during Vibrio harveyi infection in the golden pompano Trachinotus ovatus.

Dev Comp Immunol 2021 04 11;117:103959. Epub 2020 Dec 11.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China. Electronic address:

Vibrio harveyi is regarded as serious pathogen for marine fishes. However, host defense mechanisms involved in V. harveyi infection remain incompletely defined. The transcription factor IFN regulatory factor 7 (IRF7) is largely associated with host defense against viral infections, and the role of IRF7 during V. harveyi infection in fish has not been well illuminated previously. In this study, IRF7 from golden pompano (Trachinotus ovatus) was characterized (TroIRF7). The TroIRF7 gene is 1323 bp, which encodes 440 amino acid residues. Multiple amino acid alignments of TroIRF7 shows 30.37%-80.18% identity with other fish IRF7s, including Epinephelus coioides (80.18%), Larimichthys crocea (79.72%), Collichthys lucidus (79.26%), Miichthys miiuy (79.26%), Channa argus (78.77%), Cynoglossus semilaevis (72.67%), and Gadus morhua (65.23%). Like other IRF7s, TroIRF7 also contains 3 conserved domains: an N-terminal DNA-binding domain (DBD), an IRF association domain (IAD), and a C-terminal serine-rich domain (SRD). In the DBD, 4-5 conserved tryptophans were observed, which is a characteristic unique to all fish IRF7 members. TroIRF7 was constitutively expressed, with high levels in gill, head kidney, spleen, skin, and intestine. V. harveyi infection-induced TroIRF7 transcripts significantly up-regulation and translocation to the nucleus. TroIRF7 overexpression promote the fish to inhibit the replication of V. harveyi. And TroIRF7 knockdown led to decreased bacterial clearance in fish tissue. Furthermore, over-expression of TroIRF7 resulted in an increased production of interferon a3 and IFN signaling molecule in the spleen, suggesting that V. harveyi activates the IRF7- IFN pathway. These results suggest that TroIRF7 is an important component of immune responses against V. harveyi infection.
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http://dx.doi.org/10.1016/j.dci.2020.103959DOI Listing
April 2021

Molecular characterization and antibacterial immunity functional analysis of liver-expressed antimicrobial peptide 2 (LEAP-2) gene in golden pompano (Trachinotus ovatus).

Fish Shellfish Immunol 2020 Nov 3;106:833-843. Epub 2020 Sep 3.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, Hainan, 570228, PR China; Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province, Haikou, Hainan, 570228, PR China. Electronic address:

Liver-expressed antimicrobial peptide-2 (LEAP-2) is a member of the antimicrobial peptides family. Research has demonstrated that LEAP-2 contains a number of cations and plays a key role in the innate immune system of organism. In this study, we cloned and identified TroLEAP-2, from the golden pompano (Trachinotus ovatus), and analyzed its functions in vivo and in vitro. Results showed that TroLEAP-2 contains a 321 bp open reading frame (ORF) that encodes 106 putative amino acids with a molecular weight of 11.65 kDa. The mature TroLEAP-2 peptide possesses four conserved cysteine residues, which can form a core structure with two disulfide bonds between the cysteine residues in the relative 1-3 (Cys 77 and Cys 88) and 2-4 (Cys 83 and Cys 93) positions. It has a high amino acid sequence similarity (38.68%-83.02%) with the liver-expressed antimicrobial peptide -2 of other teleosts. Phylogenetic analysis showed that TroLEAP-2 clustered with the LEAP-2 of Paralichthys olivaceus and Miichthy milluy. TroLEAP-2 was most abundantly expressed in the liver, spleen, and kidney, and was significantly upregulated during Edwardsiella tarda and Streptococcus agalactiae infection. Purified recombinant TroLEAP-2 (rTroLEAP-2) could significantly inhibit the in vitro growth of E. tarda and S. agalactiae. Overexpression of TroLEAP-2 in vivo was shown to significantly reduce E. tarda and S. agalactiae colonization of tissues, whereas its knockdown resulted in an increase of bacteria in fish tissues. We also saw that TroLEAP-2 overexpression significantly improved macrophage activation in vivo. Moreover, TroLEAP-2 can induce the expression of nonspecific immune-related genes. These results showed that it might play a significant role in the innate immune system of golden pompano. In conclusion, our results indicate that TroLEAP-2 plays an important role in antibacterial immunity and provides a new avenue for protection against pathogenic infections in golden pompano.
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http://dx.doi.org/10.1016/j.fsi.2020.09.002DOI Listing
November 2020

Establishment and characterization of a new cell line from the muscle of humpback grouper (Cromileptes altivelis).

Fish Physiol Biochem 2020 Dec 25;46(6):1897-1907. Epub 2020 Jun 25.

Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province, Haikou, 570228, Hainan, People's Republic of China.

The humpback grouper (Cromileptes altivelis) is a commercially important species of the family Epinephelidae. With the development in aquaculture industry, C. altivelis breeding has gradually increased in volumetric production, leading to the occurrence of various diseases. In this study, we established a new cell line (CAM) derived from the muscle tissue of C. altivelis. Our results showed that the optimal growth temperature and working concentration of fetal bovine serum (FBS) of CAM cells were 28 °C and 15%, respectively. DNA sequencing and comparative analysis of 18S rRNA gene sequence showed that CAM cell line was originated from C. altivelis. Chromosome analysis showed that the modal chromosome number of CAM cells was 48. After transfection using pEGFP-N3 plasmid, CAM cells exhibited high transfection efficiency, indicating that CAM cells could be used in foreign gene expression studies. Further, cytotoxicity analysis revealed that CAM cells were sensitive to Vibrio harveyi and Edwardsiella tarda. Moreover, the cytotoxicity of heavy metals (Hg, Cd, and Cu) to CAM cells was dose-dependent. This CAM cell line might be used as an ideal tool in vitro for analyzing and understanding the mechanisms of pathogenesis, host-pathogen interactions, and toxicity assay of heavy metals.
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http://dx.doi.org/10.1007/s10695-020-00841-5DOI Listing
December 2020

Insulin-like growth factor binding protein 3 gene of golden pompano (TroIGFBP3) promotes antimicrobial immune defense.

Fish Shellfish Immunol 2020 Aug 8;103:47-57. Epub 2020 Apr 8.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, Hainan, 570228, PR China; Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province, Haikou, Hainan, 570228, PR China. Electronic address:

Insulin-like growth factor binding protein 3 (IGFBP3), an important member of the IGFBP family, plays an important biological role in regulating cellular proliferation, differentiation, growth, apoptosis, and innate immunity. However, studies concerning IGFBP3 in teleosts are very limited and IGFBP3 function remains unclear. In this study, we conducted both in vivo and in vitro functional analyses of an IGFBP3 (TroIGFBP3) from the teleost fish golden pompano (Trachinotus ovatus). TroIGFBP3 is composed of 286 amino acid residues and shares a high amino acid sequence similarity (50.18%-93.71%) with other IGFBP3 sequences in humans and teleosts. TroIGFBP3 was widely distributed in various tissues, with the highest expression in the liver. TroIGFBP3 expression was significantly upregulated following Vibrio harveyi infection. The results of in vitro assays showed that TroIGFBP3 could stimulate macrophage activation and promote peripheral blood leukocytes (PBLs) proliferation. Meanwhile, TroIGFBP3 overexpression significantly inhibited bacterial infection in fish tissues, whereas TroIGFBP3 knockdown resulted in increased bacterial dissemination and colonization in golden pompano tissues in vivo. Furthermore, recombinant TroIGFBP3 could inhibit cellular proliferation and promote apoptosis of mouse tumor cells. Taken together, these results indicated that TroIGFBP3 plays a significant role in innate antibacterial immunity and provides a theoretical foundation for investigating the function of IGFBP3 in fish immune response.
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http://dx.doi.org/10.1016/j.fsi.2020.04.002DOI Listing
August 2020

Construction and analysis of the immune effect of Vibrio harveyi subunit vaccine and DNA vaccine encoding TssJ antigen.

Fish Shellfish Immunol 2020 Mar 27;98:45-51. Epub 2019 Dec 27.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China. Electronic address:

Vibrio harveyi, a severe pathogen infects different kinds of sea animals, causes huge economic loss in aquaculture industry. In order to control the Vibriosis disease caused mainly by V. harveyi and other Vibrio spp., the best solution lies in developing corresponding efficient vaccines. In this study, we have cloned and analysed a putative antigen TssJ from the T6SS of V. harveyi, which has the potential as a vaccine against infection. The sequence analysis and western blotting experiments indicated that TssJ anchored in outer membrane and there were several antigenic determinants existed on its extracellular region. Two forms of universal vaccines, subunit vaccine and DNA vaccine, were developed based on TssJ and applied in Trachinotus ovatus. The results showed that both of the two vaccines could generate a moderate protection in fish against V. harveyi. The relative percentage survival (RPS) of subunit vaccine and DNA vaccine were 52.39% and 69.11%, respectively. Immunological analysis showed both subunit vaccine and DNA vaccine enhanced acid phosphatase, alkaline phosphatase, superoxide dismutase, and lysozyme activities. Specific serum antibodies against TssJ in the fish vaccinated with subunit vaccine was much higher than that in the DNA vaccine group. Several immune-related genes, i.e., IL10, C3, MHC Iα, MHC IIα, and IgM, were induced both by the two forms of vaccines. TNFα and Mx were only upregulated in the DNA vaccine group. However, the induction levels of these genes induced by DNA vaccine were higher than subunit vaccine. All these findings suggested that TssJ from V. harveyi had a potential application value in vaccine industry.
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http://dx.doi.org/10.1016/j.fsi.2019.12.079DOI Listing
March 2020

Evaluation of Lactococcus lactis HNL12 combined with Schizochytrium limacinum algal meal in diets for humpback grouper (Cromileptes altivelis).

Fish Shellfish Immunol 2019 Nov 25;94:880-888. Epub 2019 Sep 25.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China. Electronic address:

The humpback grouper (Cromileptes altivelis) is a commercially valuable species of the family Epinephelidae; however, its marketization suffers from slow growth speed, low survival rate, and various pathogenic diseases. Lactococcus lactis and Schizochytrium limacinum are commonly used as immunostimulants due to their health benefits for the aquatic organisms. In the present study, we assessed the effects of dietary supplementation with L. lactis HNL12 combined with S. limacinum algal meal on the growth performances, innate immune response, and disease resistance of C. altivelis against Vibrio harveyi. The results showed that fish fed with a combination diet of L. lactis and S. limacinum exhibited significantly higher final weight, percent weight gain, and specific growth rate compared with groups fed with them alone. A bacterial challenge experiment indicated that the group fed with the L. lactis combined with S. limacinum diet achieved the highest relative percent of survival value (68.63%), suggesting that L. lactis and S. limacinum significantly improved the disease resistance against V. harveyi after a 4-week feeding trial. Moreover, the respiratory burst activity of macrophages of fish fed with a L. lactis combined with S. limacinum diet was significantly higher than that of fish fed the control diet after 1, 2, and 3 weeks of feeding. The serum superoxide dismutase of fish fed with a L. lactis combined with S. limacinum diet significantly increased compared to those fed the control diet after 1 and 2 weeks of feeding, while the serum alkaline phosphatase of fish fed with a L. lactis combined with S. limacinum diet after 2 and 4 weeks was significantly increased, compared to the control group. The serum lysozyme activities of fish fed with a L. lactis combined with S. limacinum diet significantly increased compared to the control group after 2 weeks of feeding. Furthermore, transcriptome sequencing of the C. altivelis head kidney was conducted to explore the immune-regulating effects of the L. lactis combined with S. limacinum diet on C. altivelis. A total of 86,919 unigenes, annotated by at least one of the reference databases (Nr, Swiss-Prot, GO, COG, and KEGG), were assembly yielded by de novo transcriptome. In addition, 157 putative differentially expressed genes (DEGs) were identified between the L. lactis combined with S. limacinum group and the control group. For pathway enrichment, the DEGs were categorized into nine KEGG pathways, which were mainly related to infective diseases, antigen processing and presentation, digestive system, and other immune system responses. The findings of this study suggest that the L. lactis combined with S. limacinum diet can induce positive effects on the growth, immunity, and disease resistance of C. altivelis against V. harveyi. This study expands our understanding of the synergistic combinations of probiotics and prebiotics in aquaculture.
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http://dx.doi.org/10.1016/j.fsi.2019.09.059DOI Listing
November 2019

Comparative analysis of the expression patterns of IL-1β, IL-11, and IL-34 in golden pompano (Trachinotus ovatus) following different pathogens challenge.

Fish Shellfish Immunol 2019 Oct 15;93:863-870. Epub 2019 Aug 15.

Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China.

Interleukins (ILs) are a subgroup of cytokines, which are molecules involved in the intercellular regulation of the immune system. These cytokines have been extensively studied in mammalian models, but systematic analyses of fish are limited. In the current study, 3 IL genes from golden pompano (Trachinotus ovatus) were characterized. The IL-1β protein contains IL-1 family signature motif, and four long helices (αA - αD) in IL-11 and IL-34, which were well conserved. All 3 ILs clustered phylogenetically with their respective IL relatives in mammalian and other teleost species. Under normal physiological conditions, the expression of IL-1β, IL-11, and IL-34 were detected at varied levels in the 11 tissues examined. Most of the 3 ILs examined were highly expressed in liver, spleen, kidney, gill, or skin. Following pathogenic bacterial, viral, or parasitic challenge, IL-1β, IL-11, and IL-34 exhibited distinctly different expression profiles in a time-, tissue-, and pathogen-dependent manner. In general, IL-1β was expressed at higher levels following challenge with all pathogens examined than was observed for IL-11 and IL-34. Furthermore, Streptococcus agalactiae and Cryptocaryon irritans caused higher levels of IL-1β and IL-11 expression than Vibrio harveyi and viral nervous necrosis virus (VNNV). The increased expression of IL-34 caused by VNNV and C. irritans were higher than that caused by V. harveyi and S. agalactiae. These results suggest that these 3 ILs in T. ovatus may play different effect pathogen type specific responses.
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http://dx.doi.org/10.1016/j.fsi.2019.08.018DOI Listing
October 2019

Construction and characterization of a DNA vaccine encoding the SagH against Streptococcus iniae.

Fish Shellfish Immunol 2019 Jun 24;89:71-75. Epub 2019 Mar 24.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China.

Streptococcus iniae is an important aquaculture pathogen that is associated with disease outbreaks in wild and cultured fish species. Streptolysin S has been identified as an important virulence factor of S. iniae. With an aim to develop effective vaccines against S. iniae for Japanese flounder (Paralichthys olivaceus), in this study, we constructed a DNA vaccine based on the sagH gene, which belongs to the streptolysin S-associated gene cluster. In fish vaccinated with pSagH, the transcription of sagH was detected in tissues and SagH protein was also detected in the muscles of pSagH-vaccinated fish by immunohistochemistry. The immunoprotective effect of SagH showed that fish vaccinated with pSagH at one and two months exhibited a high relative percent survival (RPS) of 92.62% and 90.58% against S. iniae serotype I, respectively. In addition, SagH conferred strong cross protection against S. iniae serotype II and resulted in an RPS of 83.01% and 80.65% at one and two months, respectively. Compared to the control group, fish vaccinated with pSagH were able to induce much stronger respiratory burst activity, and higher titer of specific antibodies. The results of quantitative real-time PCR demonstrated that pSagH upregulated the expression of several immune genes that are possibly involved in both innate and adaptive immune responses. These results indicate that pSagH is a candidate DNA vaccine candidate against S. iniae serotype I and II infection in Japanese flounder in aquaculture.
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http://dx.doi.org/10.1016/j.fsi.2019.03.045DOI Listing
June 2019

TroCCL4, a CC chemokine of Trachinotus ovatus, is involved in the antimicrobial immune response.

Fish Shellfish Immunol 2019 Mar 3;86:525-535. Epub 2018 Dec 3.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China.

CC chemokines are a large subfamily of chemokines that play an important role in the innate immune system. To date, several CC chemokines have been identified in fish species; however, the activities and functions of these putative chemokines remain ambiguous in teleosts, especially in the golden pompano, Trachinotus ovatus. Here, we characterized CC chemokine ligand 4 from T. ovatus (TroCCL4) and studied its functions. TroCCL4 contains a 294 bp open reading frame that encodes a putative peptide comprising 97 amino acids. TroCCL4 shares a high amino acid sequence similarity of 31.11%-78.35% with other CC chemokines sequences in humans and teleosts and has four cysteine residues that are conserved among other CC chemokines. TroCCL4 is also related to the macrophage inflammatory protein (MIP) group of CC chemokines. TroCCL4 expression was most abundant in immune organs and significantly upregulated in a time-dependent manner following Edwardsiella tarda infection. Recombinant TroCCL4 (rTroCCL4) induced the migration of peripheral blood leukocytes and the cellular proliferation of head kidney lymphocytes. In addition, rTroCCL4 inhibited the growth of Escherichia coli and E. tarda, indicating an antimicrobial function. Furthermore, the results of in vivo analysis showed that TroCCL4 overexpression in T. ovatus significantly enhanced macrophage activation; upregulated the gene expression of interleukin 1-β (IL-1β), interleukin 15 (IL15), interferon-induced Mx protein (Mx), tumor necrosis factor α (TNFα), complement C3, and major histocompatibility complex (MHC) class Iα and class IIα; and protected against bacterial infection in fish tissues. In contrast, knockdown of TroCCL4 expression resulted in increased bacterial dissemination and colonization in fish tissues. Taken together, our results provide evidence indicating that TroCCL4 has the ability to stimulate leukocytes and macrophages and enhance host immunity to defend against bacterial infection.
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http://dx.doi.org/10.1016/j.fsi.2018.11.080DOI Listing
March 2019

A β-defensin gene of Trachinotus ovatus might be involved in the antimicrobial and antiviral immune response.

Dev Comp Immunol 2019 03 15;92:105-115. Epub 2018 Nov 15.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, PR China; Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, PR China.

Defensins are a group of small cationic and cysteine-rich peptides that are important components of the innate immune system. However, studies on defensins in teleosts are very limited, particularly studies on defensin functions through in vivo assays. In this study, we cloned and identified one β-defensin (TroBD) the golden pompano, Trachinotus ovatus, and analyzed the functions of TroBD in both in vivo and in vitro assays. TroBD is composed of 63 amino acids and shares high sequence identities (27.27-98.41%) with known β-defensins of other teleosts. The protein has a signature motif of six conserved cysteine residues within the mature peptide. The expression of TroBD was most abundant in the head kidney and spleen and was significantly upregulated following infection by Vibrio harveyi and viral nervous necrosis virus (VNNV). Purified recombinant TroBD (rTroBD) inhibited the growth of V. harveyi, and its antimicrobial activity was influenced by salt concentration. TroBD was found to have a chemotactic effect on macrophages in vitro. The results of an in vivo study demonstrated that TroBD overexpression/knockdown in T. ovatus significantly reduced/increased bacterial colonization or viral copy numbers in tissues. Taken together, these results indicate that TroBD plays a significant role in both antibacterial and antiviral immunity and provide new avenues for protection against pathogen infection in the aquaculture industry.
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http://dx.doi.org/10.1016/j.dci.2018.11.011DOI Listing
March 2019

Effects of dietary administration of Lactococcus lactis HNL12 on growth, innate immune response, and disease resistance of humpback grouper (Cromileptes altivelis).

Fish Shellfish Immunol 2018 Nov 18;82:296-303. Epub 2018 Aug 18.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China. Electronic address:

Lactic acid bacteria are a common group of probiotics that have been widely studied and used in aquaculture. In the present study, we isolated Lactococcus lactis HNL12 from the gut of wild humpback grouper (Cromileptes altivelis) and explored its probiotic properties. For this purpose, L. lactis HNL12 was added to the commercial fish feed. The results showed that HNL12 had high auto-aggregation ability and strong tolerance to simulated gastrointestinal stress. When C. altivelis consumed a diet containing 0 (control), 10, 10, or 10 CFU/g HNL12 for four weeks, all of the groupers fed a diet with HNL12 had significantly increased percent weight gain (PWG), especially those fed with 10 CFU/g, which had a PWG of 231.45%. Compared to the control, fish fed with L. lactis HNL12 exhibited significantly increased survival rates following injection with Vibrio harveyi after one month. Immunological analysis showed that C. altivelis fed with HNL12 had (i) enhanced respiratory burst activity of head kidney macrophages, superoxide dismutase, acid phosphatase, and lysozyme activities of serum; (ii) an improved survival rate from 36% to 70%; and (iii) upregulated expression of a broad spectrum of immunity. Meanwhile, de novo transcriptome assembly yielded 89,314 unigenes, which were annotated by at least one of the reference databases (Nr, Swiss-Prot, GO, COG and KEGG). A total of 307 genes showed significantly different expression between the groups fed with or without added HNL12. GO and KEGG enrichment analyses of the significantly different expression gene categories and pathways were related to infectious diseases, antigen processing and presentation, and other immune system responses. These results indicate that L. lactis HNL12 is effective for enhancing the growth, immunity, and disease resistance of C. altivelis; this study also provides insight into the use of probiotics for commercial applications.
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http://dx.doi.org/10.1016/j.fsi.2018.08.039DOI Listing
November 2018

MHC class IIα polymorphism and its association with resistance/susceptibility to Vibrio harveyi in golden pompano (Trachinotus ovatus).

Fish Shellfish Immunol 2018 Sep 12;80:302-310. Epub 2018 Jun 12.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, PR China. Electronic address:

The major histocompatibility complex (MHC) plays an important role in the vertebrate immune response to antigenic peptides, and it is essential for recognizing foreign pathogens in organisms. In this study, MHC class IIα (Trov-MHC IIα) from the golden pompano (Trachinotus ovatus) was first cloned and identified. The gene structure of Trov-MHC IIα was contained four exons and three introns. High levels of polymorphism were found in the exon 2 of Trov-MHC IIα. A total of 29 different MHC class IIα alleles with high polymorphism were identified from 80 individuals. The ratio of non-synonymous substitutions (dN) to synonymous substitutions (dS) was 3.157 (>1) in the peptide binding regions (PBRs) of Trov-MHC IIα, suggesting positive balancing selection. Six alleles were selected to analyze the association between alleles and resistance/susceptibility to Vibrio harveyi in golden pompano. The results showed that Trov-DAA*6401 and Trov-DAA*6702 alleles were associated with the resistance to V. harveyi in golden pompano, while alleles Trov-DAA*6304 and Trov-DAA*7301 were associated with the susceptibility to V. harveyi in golden pompano. This study confirmed the association between alleles of MHC class IIα and disease resistance, and also detected some alleles which might be correlated with high V. harveyi-resistance. These disease resistance-related MHC alleles could be used as potential genetic markers for molecular marker-assisted selective breeding in the golden pompano.
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http://dx.doi.org/10.1016/j.fsi.2018.06.020DOI Listing
September 2018

Transcriptome analysis provides insights into the immune responsive pathways and genes in the head kidney of tiger grouper (Epinephelus fuscoguttatus) fed with Spatholobus suberectus, Phellodendron amurense, or Eclipta prostrata.

Fish Shellfish Immunol 2018 Feb 6;73:100-111. Epub 2017 Dec 6.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou, Hainan, 570228, PR China; Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, Haikou, Hainan, 570228, PR China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Sciences, Hainan University, Haikou, Hainan 570228, PR China. Electronic address:

The tiger grouper, Epinephelus fuscoguttatus, is an economically important fish in Southeast Asia but has been plagued by several diseases. Spatholobus suberectus (S), Phellodendron amurense (P), and Eclipta prostrate (E) are three commonly used Chinese medicinal herbs. Although previous pharmacological and clinical studies indicated that S, P, and E possess a variety of beneficial functions in mammals, little is known about their functions in farmed fish and the underlying molecular mechanism of their actions. Challenge tests in this study showed that after 14 days of diet supplement, all these herbs could effectively enhance the disease resistance of E. fuscoguttatus against Vibrio harveyi. However, the non-specific immune parameters of the herb-supplemented groups were not significantly different from the control group. To further explore the molecular mechanism of herbal immune-regulating effects on E. fuscoguttatus, transcriptome sequencing and RNA-Seq technique were applied on E. fuscoguttatus kidney. De novo transcriptome assembly of E. fuscoguttatus kidney yield 80,014 unigenes, among which, 44,901 (56.12%) were annotated with at least one of the public databases (Nr, Nt, Swiss-Prot, KEGG, COG, GO). Among these, 22,738, 11,700 and 27,457 unigenes were assigned to 57, 25 and 258 categories of GO, COG and KEGG databases, respectively. Using Solexa/Illumina's DGE platform, a total of 231, 186 and 144 putative differentially expressed genes (DEGs) were detected in P, E and S group compared with the control group. GO analysis indicated that in P and E, down-regulated DEGs were dominant in almost every GO term; whereas in S, up-regulated DEGs were more dominant. KEGG pathway analysis revealed that putative DEGs in all three herb groups were obviously enriched in the pathways related to infective diseases and immune system. We also identified a number of immune relative genes and pathways (TLR5, IL8 and MAPK pathway, for instance) associated with P, E and S's regulatory effects on E. fuscoguttatus. This study will enrich the E. fuscoguttatus transcriptome database, contribute to a better understanding of the molecular mechanisms associated with the immunoregulatory activities of Chinese medicinal herbs on teleost and provide valuable information on the prevention of grouper Vibrio diseases using Chinese medicinal herbs.
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http://dx.doi.org/10.1016/j.fsi.2017.12.004DOI Listing
February 2018

Identification, polymorphism and expression of MHC class Iα in golden pompano, Trachinotus ovatus.

Fish Shellfish Immunol 2017 Aug 26;67:55-65. Epub 2017 May 26.

State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, China; Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine Science, Hainan University, Haikou, Hainan 570228, China. Electronic address:

The classical major histocompatibility complex class I (MHC I) plays a vital role in the immune system. In this study, we cloned and identified golden pompano (Trachinotus ovatus) MHC Iα (Trov-MHC Iα), which encodes 351 amino acid residues including a leader peptide, α1, α2, α3 domain, a transmembrane region and a cytoplasmic domain. Twenty six different sequences, which encoded various numbers of amino acid residues ranging from 348 to 354, were obtained from 12 individuals. Highly genetic polymorphism was found in the Trov-MHC Iα, especially in the α1 and α2 domains. Meanwhile, in the α1 and α2 domains, 21 positive selected positions were revealed by site models, indicating the diversity of Trov-MHC Iα may be mainly generated by positive selection. Moreover, quantitative real-time reverse transcription PCR and western blotting analyses demonstrated that Trov-MHC Iα was ubiquitously expressed in the nine tested tissues and more highly expressed in intestine, head kidney, gill, and spleen. In the head kidney and spleen, Trov-MHC Iα was significantly upregulated under LPS or poly I:C stimulation. The results of this study provide valuable insight into molecular polymorphism, evolutionary mechanism, expression and function of MHC Iα in the immune system of golden pompano.
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http://dx.doi.org/10.1016/j.fsi.2017.05.058DOI Listing
August 2017

Dietary administration of Bacillus subtilis HAINUP40 enhances growth, digestive enzyme activities, innate immune responses and disease resistance of tilapia, Oreochromis niloticus.

Fish Shellfish Immunol 2017 Jan 2;60:326-333. Epub 2016 Dec 2.

Key Laboratory of Tropical Biological Resources of Ministry of Education, College of Marine Sciences, Hainan University, Haikou 570228, PR China. Electronic address:

The probiotic properties of Bacillus subtilis HAINUP40 isolated from the aquatic environment, and the effects of dietary administration of B. subtilis HAINUP40 on the growth performance, intestinal probiotic recovery, digestive enzyme activities, innate immunity and disease resistance of tilapia (Oreochromis niloticus) were evaluated. The probiotic properties investigated include tolerance to simulated gastrointestinal stress, auto-aggregation, cell surface hydrophobicity and extracellular enzyme production. The cell number of B. subtilis changed little after 4 h in simulated gastric fluid at pH = 2.0, 3.0, 4.0 and simulated intestinal fluid at pH = 6.8.B.subtilis HAINUP40 revealed strong auto-aggregation property (34.6-87.0%) after 24 h incubation period. It exhibited significant cell surface hydrophobicity in xylene (28.8%) and chloroform (41.3%) and produced extracellular proteases and amylase. After tilapia (mean weight = 95 ± 8 g) were fed with a diet containing 10 cfu/g B. subtilis HAINUP40, their final body weight, percent weight gain (PWG), specific growth rate (SGR), total antioxidant capacity (T-AOC) and serum superoxide dismutase (SOD) increased significantly (p < 0.05) after 8 weeks; feed conversion rate (FCR) is significantly lower (p < 0.05) after 8 weeks; the protease and amylase activity in the digestive tract increased significantly (p < 0.05) after 4 and 8 weeks; and respiratory bursts and serum lysozyme activity increased significantly (p < 0.05) after 2 weeks. Moreover, being challenged with pathogenic Streptococcus agalactiae for 2 weeks, the relative percent survival (RPS%) is 52.94%. The results of this study strongly suggest that dietary supplement of B. subtilis HAINUP40 can effectively enhances the growth performance, immune response, and disease resistance of Nile tilapia.
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http://dx.doi.org/10.1016/j.fsi.2016.12.003DOI Listing
January 2017

Norcantharidin enhances ABT-263-mediated anticancer activity in neuroblastoma cells by upregulation of Noxa.

Oncol Rep 2014 Aug 30;32(2):716-22. Epub 2014 May 30.

Department of Obstetrics and Gynecology, People's Hospital of Zhengzhou University, Zhengzhou, Henan 450003, P.R. China.

Neuroblastoma is an aggressive childhood disease. Even with intensive conventional treatments, the long term survival rate for children with neuroblastoma remains less than 40%, highlighting the importance of finding new therapies. Bcl-2 family proteins play crucial roles in survival, proliferation and chemotherapeutic resistance of neuroblastoma cells. Therefore, targeting Bcl-2 with small molecule inhibitor ABT-263 could be a novel strategy for treatment of neuroblastoma. However, previous studies indicated that most neuroblastoma cell lines are resistant to ABT-263-mediated apoptosis. Thus, it is crucial to discover approaches that could overcome ABT-263 resistance. In this study, we examined the anticancer activity of ABT-263 in combination with norcantharidin (NCTD), a small-molecule anticancer drug derived from a traditional Chinese medicine, in human malignant neuroblastoma cells. We found that NCTD substantially enhanced ABT-263-mediated apoptosis induction, cell viability inhibition, and clonal formation inhibition in neuroblastoma SH-SY5Y and CHLA-119 cell lines. Moreover, the combination anticancer activity was accompanied by upregulation of Noxa, and was associated with characteristics of mitochondrial apoptosis signaling, such as cytosolic release of cytochrome c, activation of caspase-9,-3, and cleavage of PARP. Notably, we observed that knockdown of Noxa significantly attenuated cell death induction by cotreatment with ABT-263 and NCTD, indicating Noxa essentially contributes to the combination anticancer effect. Collectively, our study demonstrated that NCTD could overcome ABT-263-resistance in neuroblastoma cells, and suggested that combinational treatment of ABT-263 with NCTD might be a novel therapeutic option for children with neuroblastoma.
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http://dx.doi.org/10.3892/or.2014.3228DOI Listing
August 2014

[Determination of phenylethanoid glycosides from Cistanche deserticola in spring and autumn with LC-MS].

Zhong Yao Cai 2004 Mar;27(3):175-7

Xinjiang Institute of Physic and Chemistry of Chinese Academy of Science, Urumuqi 830011.

Objective: To have a contrast study on phenylethanoid glycosides from Cistanche deserticola Y. C. Ma collected in different seasons.

Methods: LC/MS method has been applied for the analysis of four kinds of phenylethanoid glycosides compunds (echinacoside, acteoside, cistanoside A and 2'-acetylacteoside) from Cistanche deserticola Y. C. Ma in spring and autumn.

Results: According to the special MS spectra and HPLC chromatogram, this four kinds of phenylethanoid glycosides compounds were detected in each Cistanche deserticola Y. C. Ma, but the content is considerable different except the acteoside.

Conclusion: The content of phenylethanoid glycosides from Cistanche deserticola Y. C. Ma in different seasons has a difference from each other, the quality of Cistanche deserticola Y. C. Ma is also different.
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March 2004
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