1.Research progress of new isothermal amplification technology in rapid diagnosis of Plasmodium and other parasites
YANG Jun-jie ; WU Tian ; LIU Cheng-long ; LIAN Meng-ze ; ZHANG Yi-long
China Tropical Medicine 2022;22(09):884-
Abstract: At present, nucleic acid detection technology based on the PCR principle is commonly used to detect malaria parasites, the existing Plasmodium detection methods mainly include microscopy, antigen immunoassay, and nucleic acid detection,but due to the long detection time, high personnel and equipment requirements, and other shortcomings, its popularization, and application at the grassroots level are limited. What challenges previous Plasmodium detection methods are the lack of experienced professionals and advanced equipment at the grassroots as well as the requirement of rapid detection of large samples under extreme conditions. The isothermal amplification technology developed in recent years has potential application prospects due to its simplicity, rapidity, high sensitivity, and high specificity. This article attempts to review the principles, characteristics, and prospects of various isothermal amplification technologies, and on this basis, focuses on the introduction of recombinase polymerase amplification (RPA) and recombinase⁃aided isothermal amplification (RAA) assay technologies and proposes the use of such recombinant enzyme amplification technologies to achieve rapid and accurate diagnosis of common Plasmodium species possibility and imagination.
2.Analysis on genetic diversity and genetic relationship of medicinal species in Dipsacus from China by SRAP.
Da-xia CHEN ; Xue ZHANG ; Yu WANG ; Long-yun LI ; Ze ZHANG
China Journal of Chinese Materia Medica 2015;40(13):2559-2564
The author detected the genetic diversity and genetic relationship within and among eight medicinal species of Dipsacus by the approach of sequence-related amplified polymorphism (SRAP). The associated genetic parameters were calculated by POPGENE 1.31. The Genetic distance was calculated by TREECONW and the systematic diagrams of genetic relationship were clustered by UPG-MA. The results showed that, using 26 primers, a total of 558 bands were produced, of which 539 were polymorphic loci. There was a high level of genetic diversity among species (PPB = 96.59%, Na = 1.9659, Na = 1.3375, H = 0.2143, I = 0.3423). However, genetic diversity was lower within species, the average of genetic parameters was PPB = 6.97%, Na = 1.0697, Na = 1.0311, H = 0.0187, I = 0.0291. The Nei's genetic differentiation coefficient was 0.9126, indicated that most of the genetic variation existed among species. By clustering analysis, different individuals gathered in the same group and the classified result of SRAP marker between traditional modal characters was almost same. The results confirmed that SRAP marker can be used as one of the effective methods to reveal the genetic diversity and relationship among medicinal species of Dipsacus.
China
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Dipsacaceae
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classification
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genetics
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Gene Amplification
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Genetic Variation
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Plants, Medicinal
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classification
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genetics
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Polymorphism, Genetic
3.Analysis on genetic diversity among wild Dipsacus asperoides by SCoT.
Da-xia CHEN ; Xue ZHANG ; Guang-lin CUI ; Yu WANG ; Long-yun LI ; Ze ZHANG
China Journal of Chinese Materia Medica 2015;40(10):1898-1903
To investigate the genetic diversity among wild Dipsacus asperoides in China, 66 germplasmic resources of D. asperoides were analyzed by Start Codon Targeted Polymorphism (SCoT) molecular markers. Genetic distance was calculated by TREECONW software and the systematic diagram of genetic relationship was clustered by UPGMA method. The results showed that the totals of 181 bands were detected using 20 primers , among which 109 were polymorphic bands. The average percentage of polymorphic bands was 60.13%. Genetic distance changed from 0.030 6 to 0.181 4. The clustering results showed that there was no significant correlation between the classification of the wild D. asperoides and their geographical origin. The relatively high genetic diversity of D. asperoides provides the basis for breeding new varieties.
China
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DNA Primers
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genetics
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DNA, Plant
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genetics
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Dipsacaceae
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chemistry
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classification
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genetics
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Genetic Variation
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Phylogeny
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Polymorphism, Genetic
4.Progress in studies on SOD mimics.
Acta Pharmaceutica Sinica 2002;37(3):235-240
5.Effect of citric acid stimulation on salivary alpha-amylase, total protein, salivary flow rate and pH value in Pi deficiency children.
Ze-min YANG ; Long-hui CHEN ; Jing LIN ; Min ZHANG ; Xiao-rong YANG ; Wei-wen CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(2):188-192
OBJECTIVETo compare the effect of citric acid stimulation on salivary alpha-amylase (sAA), total protein (TP), salivary flow rate, and pH value between Pi deficiency (PD) children and healthy children, thereby providing evidence for Pi controlling saliva theory.
METHODSTwenty PD children were recruited, and 29 healthy children were also recruited at the same time. Saliva samples from all subjects were collected before and after citric acid stimulation. The sAA activity and amount, TP contents, salivary flow rate, and pH value were determined and compared.
RESULTS(1) Citric acid stimulation was able to significantly increase salivary flow rate, pH value, sAA activities, sAA specific activity and sAA amount (including glycosylated and non-glycosylated sAA amount) in healthy children (P<0.05), while it could markedly increase salivary flow rate, pH value, and glycosylated sAA levels in PD children (P<0.05); (2) Although there was no statistical difference in determined salivary indices between the two groups (P>0.05), salivary indices except salivary flow rate and glycosylated sAA levels decreased more in PD children. There was statistical difference in sAA activity ratio, sAA specific activity ratio, and the ratio of glycosylated sAA levels between PD children and healthy children (P<0.05).
CONCLUSIONPD children had decreased response to citric acid stimulation.
Child ; Citric Acid ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Saliva ; Salivary alpha-Amylases ; metabolism ; alpha-Amylases
6.The ability of self-face recognition and its relation to the empathy in patients with schizophrenia
He CHANG ; Yi DONG ; Kai WANG ; Zu-lun XIONG ; Feng-feng ZHANG ; Xiao-qian ZHANG ; Ai-guo ZHANG ; Long-ze ZHANG ; Mian ZHANG ; Hui LI
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(3):193-195
Objective To explore the self-face recognition and its relationship to empathy in patients with schizophrenia.Methods Sixty-two schizophrenic patients and fifty -four healthy subjects were assessed with the self-face recognition task (SFRT) and the interpersonal reactivity index-C (IRI-C).Results The SFRT reaction time in the patients group( (2188 ± 1138) ms) was significantly longer than that in the control group( ( 1152 ± 326) ms) (P < 0.01 ) ;the accuracy in the patients group ( (80 ± 16) % ) was significantly lower than that in the control group ( (88 ± 6) % ) (P < 0.01 ).The IRI-C total scores,the subscores in perspective taking,the subscores in fantasy and empathic concern of IRI-C were significantly lower in the patients group(respectively(44.82 ± 10.50),(8.98 ± 3.56),( 11.87 ± 4.38 ),( 14.73 ± 4.00) ) than those in the control group ( respectively (49.85 ± 10.28),( 10.78 ± 3.86),( 14.98 ± 6.12),( 17.39 ± 4.56) ) ; the subscore in personal distress of IRI-C in the patients group(9.37 ± 5.12) was significantly higher than those in the control group(6.52 ± 3.89) ( P< 0.01 ).There was significant positive correlation between the accuracy for self-face recognition in SFRT and the subscore in fantasy of IRI-C ( r =0.322,P < 0.05 ),the reaction time of SFRT had significantly positive correlation with the subscore in personal distress.Conclusion Schizophren patients have general impairments of self-face recognition and empathic abilities,and the self-face recognition is related to the empathic abilities.
7.Early high-volume hemofiltration treatment in severe sepsis.
Ning ZHANG ; Ze-Liang QIU ; Bao-Li CHENG ; Yue JIN ; Jun-Long XU ; Tian-Zheng LOU
Journal of Zhejiang University. Medical sciences 2010;39(5):483-486
OBJECTIVETo evaluate the effect of early application of high-volume hemofiltration treatment (HVHF) on the levels of lactic acid, pro-inflammatory cytokines and C-reactive protein (CRP) in plasma, as well as APACHE II score in patients suffering from severe sepsis.
METHODSThirty patients meeting the diagnosis of severe sepsis were enrolled in the trial within 24 hours of insults. The level of lactic acid, interleukin-6 (IL-6) and CRP in plasma were measured before HVHF and at 24, 48 or 72 h following HVHF treatment.
RESULTThe plasma levels of lactic acid and IL-6 decreased significantly at 24 h, 48 h, 72 h after HVHF (P <0.05), while, IL-10 did not differ significantly following HVHF (P>0.05), when compared with that before HVHF.
CONCLUSIONThe early application of HVHF could clear the plasma lactic acid and pro-inflammatory cytokines, and improve the tissue oxygenation in severe sepsis.
APACHE ; Adult ; C-Reactive Protein ; analysis ; Female ; Hemofiltration ; methods ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Lactic Acid ; blood ; Male ; Middle Aged ; Sepsis ; blood ; therapy ; Treatment Outcome ; Young Adult
8.Construction of recombinant adenovirus co-expression vector carrying the human transforming growth factor-beta1 and vascular endothelial growth factor genes and its effect on anterior cruciate ligament fibroblasts.
Xue-Lei WEI ; Lin LIN ; Yu HOU ; Xin FU ; Ji-Ying ZHANG ; Ze-Bin MAO ; Chang-Long YU
Chinese Medical Journal 2008;121(15):1426-1432
BACKGROUNDRemodeling of the anterior cruciate ligament (ACL) graft usually takes longer than expected. Gene therapy offers a radical different approach to remodeling of the graft. In this study, the internal ribosome entry site (IRES) sequence was used to construct a new recombinant adenovirus which permits co-expression of transforming growth factor-beta1 (TGFbeta1) and vascular endothelial growth factor 165 (VEGF165) genes (named Ad-VEGF165-IRES-TGFbeta1). We investigated the effects of the new adenovirus on the migration of and matrix synthesis by ACL fibroblasts.
METHODSAdenoviral vector containing TGFbeta1 and VEGF165 genes was constructed. ACL fibroblasts were obtained from New Zealand white rabbits. After ACL fibroblasts were exposed to Ad-VEGF165-IRES-TGFbeta1, the expression of VEGF165 and TGFbeta1 proteins were assessed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Bioassay of VEGF165 and TGFbeta1 proteins were assessed by Western blotting analysis. Proliferation and migration of ACL fibroblasts were assessed by in vitro wound closure assay. Gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers were assessed by real-time PCR.
RESULTSThe results showed the successful construction of a recombinant co-expression adenovirus vector containing TGFbeta1 and VEGF165 genes. Co-expression of TGFbeta1 and VEGF165 can induce relatively rapid and continuous proliferation of ACL fibroblasts and high gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers.
CONCLUSIONCo-expression of TGFbeta1 and VEGF165 genes has more powerful and efficient effects on the migration of and matrix synthesis by ACL fibroblasts.
Adenoviridae ; genetics ; Animals ; Anterior Cruciate Ligament ; cytology ; metabolism ; Cell Movement ; Cells, Cultured ; Collagen ; genetics ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibroblasts ; physiology ; Fibronectins ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing
9.In vitro osteoclast-suppressing effect of sodium ibandronate.
Wei ZHANG ; Da-long YANG ; Yun-xia WANG ; Hui-wang WANG ; Zeng-jiang ZHEN ; Ying-ze ZHANG ; Yong SHEN
Chinese Medical Journal 2013;126(4):751-755
BACKGROUNDBisphosphonates (BPs) have been reported to reduce local recurrence in giant cell tumor (GCT) of bone because of their osteoclast-suppressing effect; however, the optimal mode of delivery and the dose and duration of treatment of BPs remain to be established. To address these issues, it is first necessary to clarify the manner of action of BPs on osteoclasts. We herein evaluated the osteoclast-suppressing effect of sodium ibandronate in vitro.
METHODSMouse osteoclasts (OCLs) were generated in vitro using mouse bone marrow mononuclear cells. First, various concentrations of sodium ibandronate and equal amounts of phosphate-buffered saline were added to cell culture media. The number of multinucleated cells (over three nuclei) was recorded in each group, OCL formation was compared, and the most effective concentration of sodium ibandronate was determined. Then, high concentrations of sodium ibandronate were added to the experimental cell culture media; no ibandronate was given in the control group. Comparisons were made between the two groups in terms of OCL adhesion, migration, and bone resorption.
RESULTSOCL formation was suppressed by sodium ibandronate in vitro; the most pronounced effect was observed at the concentration of 10(-5) mol/L. OCL migration and bone resorption were significantly suppressed at this concentration, though there was no effect on OCL adhesion.
CONCLUSIONSSodium ibandronate was effective in suppressing OCLs and decreasing resorption in GCT. The strong anti-OCL effectiveness at a high concentration in vitro indicates a topical mode of application.
Animals ; Bone Resorption ; Cell Movement ; drug effects ; Cells, Cultured ; Diphosphonates ; pharmacology ; Mice ; Osteoclasts ; cytology ; drug effects
10.Analysis of axial symptoms after indirect decompression for ossification of the posterior longitudinal ligament of the cervical spine.
Hui WANG ; Wen-yuan DING ; Yong SHEN ; Ying-ze ZHANG ; Wei ZHANG ; Da-long YANG ; Ya-peng SUN ; Lin-feng WANG ; Lai-zhen CAO ; Lei MA
Chinese Journal of Surgery 2012;50(7):601-606
OBJECTIVETo study the etiology and clinical significance of axial symptoms after posterior operative procedures for ossification of the posterior longitudinal ligament (OPLL).
METHODSFrom February 2005 to February 2010, 76 patients with OPLL treated were retrospectively experienced. There were 34 male and 42 female with average of 52.1 years (range from 37 to 74 years), the average duration of the disease was 32.1 months (range from 11 to 56 months). Nineteen patients underwent traditional laminectomy in group A, 33 patients received open-door laminoplasty in group B and 24 patients underwent lateral mass screw fixation in group C. All patients underwent X-ray examination pre- and post operative, computed tomography were used for diagnosis of OPLL, the recovery rate was calculated using pre- and postoperative Japanese Orthopedic Association (JOA) scores for each patient. Pre- and postoperative cervical curvature index and axial symptoms were measured and compared. χ(2) test and SNK test were used as statistical methods.
RESULTSAll patients were followed up for 14 - 35 months, average (21 ± 5) months. Loss of cervical curvature index was 4.2% ± 1.7% in group A, 2.9% ± 2.2% in group B and 2.3% ± 1.9% in group C. The difference was significant in loss of cervical curvature indice between group A and B (q = 2.94, P < 0.01), group A and C (q = 4.23, P < 0.01). The average JOA recovery rate was 58.3% for group A, 64.3% for group B and 66.7% for group C. There was no significant difference in JOA recovery rate among the three groups (P > 0.05). The rate of early evident axial symptoms was 7/19 in group A, 30.3% in group B and 33.3% in group C and the difference was not statistically significant (P > 0.05). The incidence of late evident axial symptoms was 5/19 in group A, 12.1% in group B and 8.3% in group C, the difference was not significant between group B and C (χ(2) = 13.762, P < 0.01), but of statistical difference between group A and B(χ(2) = 6.368, P < 0.01), group A and C (χ(2) = 11.481, P < 0.01). No kyphotic deformity in the group A, no "Close Door" phenomenon in group B and no internal failure in group C.
CONCLUSIONThe incidence of early axial symptoms are of no significant difference among the three groups, but late axial symptoms are higher in the laminectomy than other groups, which may be associated with loss of cervical lordosis.
Adult ; Decompression, Surgical ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Laminectomy ; adverse effects ; methods ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; surgery ; Postoperative Complications ; Retrospective Studies