1.Co-expression of GRP78 and Myocilin in trabecular meshwork cells of primary open angle glaucoma
Fang, CHAI ; Hua, AI ; Juan, LI ; Xinguang, YANG
Chinese Journal of Experimental Ophthalmology 2017;35(4):300-305
Background Misfolding of Myocilin protein plays an important role in the pathogenesis of primary open angle glaucoma (POAG).GRP78 can transfer the misfolded proteins out endocytoplasmic reticulum and keep the protein synthesis function of endoplasmic reticulum.However the relationship between GRP78 and Myocilin in the pathogenesis of POAG has not been elucidated.Objective This study was to investigate and analyze the coexpressions of GRP78 and Myolicin in trabecular meshwork cells (TMCs) of POAG.Methods The trabecular meshwork tissues were obtained during the surgery of POAG and healthy donor for the isolated and in vitro culture of TMCs,and the morphology of the cells was compared between POAG-derived TMCs and donor-derived TMCs.The expression of specific factors for TMCs were detected by immunochemistry.The cells were divided into normal control group,tunicamycin (Tm)-treated group and staurosporine (STS)-treated group,and the cells were cultured by regular medium,the 1 μ mol/L Tm medium or 0.1 μmol/L STS medium for 24 hours respectively.The co-expression of GRP78 and Myocilin in the cells were detected using immunofluorescence assay.This study was approved by Ethics Committee of Xi'an No.4 Hospital,and written informed consent was obtained from each patient before surgery.Results Primarily cultured cells showed the fiat star-like and irregular appearance with 3-5 processes,round nuclei,abundant cytoplasm and black engulf particles.The 3-7 generations of POAG-derived cells grew well,showing positive expressions of laminin (LN),fibronectin (FN),neuronal specific enolase (NSE) and Vimentin.Immunofluorescence assay showed positive expressions of GRP78 and Myocilin in the cells of the Tm group,STS group and normal control group,with the reddish and green fluorescence separately.Incomplete co-expression of GRP78 and Myocilin was seen in donor-derived TMCs,and complete co-expression of GRP78 and Myocilin was found in the POAG-derived TMCs in the normal control group.Complete co-expression of GRP78 and Myocilin was exhibited in both donor-and POAG-derived TMCs in the Tm group and STS group.In addition,accumulation of GRP78 protein around nucleus was seen in both donor-and POAG-derived TMCs in the Tm group.Conclusions GRP78 and Myocilin proteins are completely co-expressed in POAG-derived TMCs,inferring that GRP78 and Myocilin play an interaction role in the pathogenesis and development of POAG.GRP78 may play a cytoprotective role against the apoptosis of TMCs caused by endoplasmic reticulum stress.
2.Relationship between oral and gastric Hp infection and diseases of upper digestive tract in children.
Yun-guang BAO ; Yi WEI ; Ai-su LANG ; Wei-jun YU ; Ai-juan YING ; Lin-qing MU ; Hui-xian YANG
Chinese Journal of Pediatrics 2003;41(6):465-466
Adolescent
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Child
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Child, Preschool
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Female
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Gastric Mucosa
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microbiology
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Gastrointestinal Diseases
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diagnosis
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microbiology
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Helicobacter Infections
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diagnosis
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microbiology
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Helicobacter pylori
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growth & development
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isolation & purification
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Humans
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Infant
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Male
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Mouth Mucosa
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microbiology
3.Research wilt disease of Salvia miltiorrhiza and its pathogen.
Li YANG ; Zuo-Qing MIAO ; Guang YANG ; Ai-Juan SHAO ; Lu-Qi HUANG ; Ye SHEN ; Xue WANG ; Mei-Lan CHEN
China Journal of Chinese Materia Medica 2013;38(23):4040-4043
Salvia miltiorrhiza is a highly valued traditional chinese medicine for the treatment of atherosclerosis-related disorders in china, such as cardiovascular and cerebrovascular diseases in China. The wilt disease is serious in the culture of S. miltiorrhiza. Wilt disease cause biomass of plant shoots and roots is lessened, active components are decreased. To solve these problems, we research the pathogen causing wilt disease of S. miltiorrhiza. The suspected pathogen is identified by morphology and etiological test. The identification was further confirmed by alignment the sequences of internal transcribed spacer (ITS) amplified by PCR. Our result show the wilt disease of S. miltiorrhiza mostly occurred in July and August, which is hot and wetter. The wilt disease rate of S. miltiorrhiza continuous cropping for one year in S. miltiorrhiz stubble is 10%, but the wilt disease rate of S. miltiorrhiza continuous cropping for three years in S. miltiorrhiz stubble is 60%-70%. The root rot of S. miltiorrhiz caused by the wilt disease, so the wilt disease was mistaken for the rot root in production. Morphological characteristics show the pathogen is Fusarium oxysporum. The sequence of ITS wes determined and found by BLAST shared 99% identity to that of F. oxysporum f. sp. cucumerinum. So it comes to the conclusion that the causing agent of wilt disease on S. miltiorrhiza belongs to F. oxysporum.
DNA, Intergenic
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genetics
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Fusarium
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genetics
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isolation & purification
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physiology
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Plant Diseases
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microbiology
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Polymerase Chain Reaction
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Salvia miltiorrhiza
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microbiology
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Seasons
4.A new method for exposing the animal's heart with maintenance of natural breathing.
Yi, ZHANG ; Youxiang, PAN ; Changjin, LIU ; Ming, TANG ; Yundong, HU ; Shansong, YANG ; Liangpin, ZHANG ; Xingwu, HU ; Ai, LI ; Juan, LUO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(3):309-10
In the experimental study on the electrophysiology and functions of heart in the animal, the thorax should be opened to expose the heart, at the same time natural breathing is maintained instead of artificial breathing. The key procedure of this new method is to avoid injuring the pleural cavity, so pneumothorax can be prevented. By means of this new method some subject studies have been finished.
Electrophysiology/instrumentation
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Heart/*physiology
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*Models, Animal
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*Respiration
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Swine
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Thoracic Surgical Procedures
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Time Factors
5.Inhibition efficacy of CFB-siRNA on laser-induced choroidal neovascularization in rat
Cheng, MA ; Qing-li, SHANG ; Jing-xue, MA ; Jian, GAO ; He-ming, XIU ; Hui-juan, YAO ; Ai-qin, YANG
Chinese Journal of Experimental Ophthalmology 2012;30(4):320-325
BackgroundChoriodal neovascularization is an important ocular manifestation of angiogenesis in eyes,which derives from the choroid capillaries.Recent studies have found that complement activation is playing a key role in the laser-induced CNV.Because of the key position of CFB in the alternative pathway,bytargeting CFB and blocking the alternative pathway may provide an approach to observe the role of this alternative pathway in the generation of CNV.Objective This study was to investigate the inhibitory effect of reconstructed complement factor B (CFB)-small interfering ribonucleicacid(siRNA)on choroidal neovascularization (CNV)and its mechanism. Methods Experimental CNV was induced by laser photocoagulation in 96 eyes of 48 clean Brown Norway rats.The rats were randomly divided into 4 groups.25,50 and 75 μg B factor siRNA were injected via caudal vein on 1 day,3,5 days after photocoagulation in different dose groups,and normal saline solution was injected at the same way in experimental control group.Other 12 normal rats were used as blank control group.Fundus fluorescein angiography(FFA) was performed on 3,7,14,21,28 days after injection of CFB-siRNA and CNV was scored.The expressions of vascular endothelial growth factor(VEGF) and factor Ⅷ in choroid were detected by immunochemistry.The expressions of CFB-siRNA,VEGF,transforming growth factor β2( TGF-β2 )proteins in choroid were determined using immunochemistry in 7,14,21,28 days,and the expressions of mRNA of CFB-siRNA,VEGF,TGF-β2 were examined by reverse transcription polymerase chain reaction(RT-PCR). ResultsFFA revealed that the CNV rates in various doses of CFB-siRNA groups were significant lower than those of experimental control group in various time points(P<0.05),and those in 75 μg B factor siRNA were decreased in comparison with 25 μg B factor siRNA (P<0.05).Immunochemistry showed that the intensities of the VEGF and factor Ⅶ expression in various doses of CFB-siRNA groups were weaker than the blank control group ( P < 0.05 ).Compared with the control group,the expression of CFB reduced in 7 days,and then approached to the level near the control group.Fourteen to twenty-one days after injection of CFB-siRNA,VEGF and TGF-β2 depressions in different doses of CFB-siRNA groups were lower than blank control group( P<0.05 ).CFB expression in choroid showed the lower levels in CFB-siRNA injection group compared with blank control group in from 7 through 21 days (P<0.05).RT-PCR displayed the gradual increase of CFB mRNA and curve-like changes of VEGF and TGF-β2 with time prolong. Conclusions Recombinated CFB-siRNA can effectively inhibit laser-induced CNV by down-regulating the expression of VEGF and factor Ⅷ.Alternative pathway of complement plays an important role in the production of CNV.
6.Effects of polydatin on learning and memory and Cdk5 kinase activity in the hippocampus of rats with chronic alcoholism.
Xin-juan LI ; Yan ZHANG ; Chun-yang XU ; Shuang LI ; Ai-lin DU ; Li-bin ZHANG ; Rui-ling ZHANG
Chinese Journal of Applied Physiology 2015;31(2):117-120
OBJECTIVETo observe the effects of polydatin on learning and memory and cyclin-dependent kinase 5 (Cdk5) kinase activity in the hippocampus of rats with chronic alcoholism.
METHODSForty rats were randomly divided into 4 groups: control group, chronic alcoholism group, low and high polydatin group. The rat chronic alcoholism model was established by ethanol 3.0 g/(kg · d) (intragastric administration). The abstinence scoring was used to evaluate the rats withdrawal symptoms; cognitive function was measured by Morris water maze experiment; Cdk5 protein expression in the hippocampus was detected by immunofluorescence; Cdk5 kinase activity in the hippocampus was detected by liquid scintillation counting method.
RESULTSThe abstinence score, escape latency, Cdk5 kinase activity in chronic alcoholism group rats were significantly higher than those of control group (P < 0.05). The abstinence score, escape latency in high polydatin group rats were significantly lower than those of chronic alcoholism group (P < 0.05); Cdk5 kinase activity in high and low polydatin group rats was significantly lower than that of chronic alcoholism group( P < 0.05); immunofluorescence showed that the Cdk5 positive cells of chronic alcoholism group were significantly increased compared with control group (P < 0.05), and the Cdk5 positive cells of polydatin groups were significantly decreased compared with chronic alcoholism group ( P < 0.05).
CONCLUSIONPolydatin-reduced the chronic alcoholism damage may interrelate with regulation of Cdk5 kinase activity.
Alcoholism ; physiopathology ; Animals ; Cyclin-Dependent Kinase 5 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Hippocampus ; drug effects ; enzymology ; Learning ; drug effects ; Memory ; drug effects ; Rats ; Stilbenes ; pharmacology
7.Effect of five fungicides on growth of Glycyrrhiza uralensis and efficiency of mycorrhizal symbiosis.
Peng-ying LI ; Guang YANG ; Xiu-teng ZHOU ; Liane-yun ZHOU ; Ai-juan SHAO ; Mei-lan CHEN
China Journal of Chinese Materia Medica 2015;40(23):4591-4596
In order to obtain the fungicides with minimal impact on efficiency of mycorrhizal symbiosis, the effect of five fungicides including polyoxins, jinggangmycins, thiophanate methylate, chlorothalonil and carbendazim on the growth of medicinal plant and efficiency of mycorrhizal symbiosis were studied. Pot cultured Glycyrrhiza uralensis was treated with different fungicides with the concentration that commonly used in the field. 60 d after treated with fungicides, infection rate, infection density, biomass indexes, photosyn- thetic index and the content of active component were measured. Experimental results showed that carbendazim had the strongest inhibition on mycorrhizal symbiosis effect. Carbendazim significantly inhibited the mycorrhizal infection rate, significantly suppressed the actual photosynthetic efficiency of G. uralensis and the most indicators of biomass. Polyoxins showed the lowest inhibiting affection. Polyoxins had no significant effect on mycorrhizal infection rate, the actual photosynthetic efficiency of G. uralensis and the most indicators of biomass. The other three fungicides also had an inhibitory effect on efficiency of mycorrhizal symbiosis, and the inhibition degrees were all between polyoxins's and carbendazim's. The author considered that fungicide's inhibition degree on mycorrhizal effect might be related with the species of fungicides, so the author suggested that the farmer should try to choose bio-fungicides like polyoxins.
Fungi
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drug effects
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growth & development
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physiology
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Fungicides, Industrial
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pharmacology
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Glycyrrhiza uralensis
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chemistry
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growth & development
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microbiology
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physiology
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Mycorrhizae
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drug effects
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growth & development
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physiology
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Plant Extracts
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chemistry
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Symbiosis
;
drug effects
8.Villoglandular adenocarcinoma of cervix:a clinicopathological study.
Zheng-cao LIU ; Lu ZHENG ; Yun-long HUO ; Xiang-hong YANG ; Ai-feng GAO ; Xiu-juan CUI
Chinese Journal of Pathology 2010;39(5):338-339
Adenocarcinoma
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metabolism
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pathology
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surgery
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Adenocarcinoma, Clear Cell
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metabolism
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pathology
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Adult
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CA-125 Antigen
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metabolism
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Carcinoembryonic Antigen
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metabolism
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Cervical Intraepithelial Neoplasia
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metabolism
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pathology
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surgery
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Cystadenocarcinoma, Serous
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metabolism
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pathology
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Diagnosis, Differential
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Female
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Follow-Up Studies
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Humans
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Hysterectomy
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Ki-67 Antigen
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metabolism
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Lymph Node Excision
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Membrane Proteins
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metabolism
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Neoplasm Invasiveness
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Uterine Cervical Neoplasms
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metabolism
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pathology
;
surgery
9.Mutation analysis of p63 gene in the first Chinese family with ADULT syndrome.
Xia WANG ; Jian YANG ; Ai-lin TAO ; Wen-lin YANG ; Hong-juan ZHANG
Chinese Medical Journal 2009;122(16):1867-1871
BACKGROUNDADULT syndrome (acro-dermato-ungual-lacrimal-tooth syndrome) is a rare ectodermal dysplasia disorder known as autosomal dominant inheritance. Recent studies have linked p63 gene mutation to the development of this disease. However, the genetic characteristics of ADULT syndrome were still not well understood.
METHODSMutation analysis of p63 gene in the first Chinese ADULT syndrome family was performed using direct DNA sequencing.
RESULTSThe sequence analysis of exon 8 of p63 gene disclosed a heterozygous G>A substitution at nucleotide 893 (R298Q) in the proband. In addition, a single nucleotide polymorphism (SNP) rs16864880 in the downstream flanking region (DFR) of p63 exon 8 was also identified in this family. The proband and the paternal side including her father exhibited the C/G genotype at this position. The C/G variant frequency in the paternal was significantly higher as compared with the maternal (6/10 vs 0/6, P = 0.034).
CONCLUSIONSADULT syndrome may be caused by the p63 gene mutation, and it might have closer genetic association with the paternal side in this family.
Adult ; Asian Continental Ancestry Group ; genetics ; DNA Mutational Analysis ; Ectodermal Dysplasia ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Models, Molecular ; Mutation ; genetics ; Pedigree ; Protein Structure, Tertiary ; Trans-Activators ; chemistry ; genetics ; Transcription Factors ; Tumor Suppressor Proteins ; chemistry ; genetics
10.Discussion on applications and mechanisms of biocontrol microoganisms used for controlling medicinal plant soil-borne diseases.
Li YANG ; Mei-Lan CHEN ; Ai-Juan SHAO ; Guang YANG
China Journal of Chinese Materia Medica 2012;37(21):3188-3192
In recent years, the soil borne disease of medicinal plants becomes severely during the process of cultivation and directly endangered the production and quality of raw materials used in traditional Chinese medicine (TCM). The chemical pesticides have been constantly used to prevent and control the soil borne disease, but only a few are effective. Meanwhile, the excessive uses of chemical pesticides also lead pesticide residues in TCM, which often exceed limit of the standard, and harm the human health and cause environmental pollution. Therefore, biological control has become a hot research point for its environmental advantages. This paper mainly discussed the mechanisms of different species of microorganisms, which could control the soil borne disease of medicinal plants, from the following aspects: improving host plants' nutrient absorption, the nutrient and space competition with the pathogenic bacteria, changing the morphology and anatomical structure of roots, adjusting the host plants' endogenous hormones, restoring the balance of host rhizosphere soil microecology and activating the host plants' defense system etc. Then put forward the prospect of biocontrol agents in the future.
Medicine, Chinese Traditional
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Pest Control, Biological
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methods
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Plant Diseases
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prevention & control
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Plants, Medicinal
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Soil Microbiology