1.Selection of Cd~(2+) Binding-peptides and Their Affinities for Heavy Metal Ions
Jing-Shuang HUANG ; Chun-Yan MA ; Xin TONG ; Zhuo-Lin YI ; Liu XU ; Can-Quan MAO ;
Microbiology 1992;0(05):-
Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.
2.Quantitative fluorescent-polymerase chain reaction for rapid prenatal diagnosis of common aneuploidies
Li LIANG ; Can LIAO ; Min PAN ; Xin YANG ; Fatao LI ; Cuixing YI ; Yan LI ; Fang FU ; Dongzhi LI
Chinese Journal of Perinatal Medicine 2012;15(2):106-112
Objective To evaluate the clinical value of quantitative fluorescent polymerase chain reaction (QF-PCR) in rapid prenatal diagnosis of aneuploidies. Methods Twenty-two short tandem repeats (STR) and AMXY located on chromosome 13,18,21,X and Y were used as markers to examine 1740 samples from high risk pregnant women in Down syndrome screening and advanced maternal age(≥35 yrs) by QF-PCR.Samples were also tested by karyotype analysis and the results of the two methods were compared. Results Karyotype analysis and QF-PCR results were successfully obtained from 1690 samples. All QF-PCR reports were obtained within 48 hours after sample collection.For 1639 samples,normal results were obtained by both karyotype analysis and QF-PCR.Among 51 samples that were found abnormal by karyotype analysis,41 were abnormal in QF-PCR.The rapid tests found all numerical abnormalities involving chromosome 21,18,13,X and Y in prenatal diagnosis,including trisomy 21 (n =30),trisomy 18 (n =6),45,XO (n =1 ),47,XYY (n=1),47,XXX (n=1),69,XXX (n=1) and mosaic 47,XXY[94]/46,XX[6] (n=1)(47,XXY in QF-PCR).No false positive results were found.The results obtained by QF-PCR were consistent with those of cytogenetic studies in 99.4% of the samples (1680/1690).Only ten cases of mosain and structural abnormality could not be found (0.6%,10/1690) by QF-PCR. Conclusions Rapid QF-PCR test might diagnose all aneuploidies involving chromosome 21,18,13,X and Y.It could provide rapid and accurate diagnosis for 99.4% pregnant women with positive Down syndrome screening and advanced maternal age.
3.Detection of Haptoglobin by Surface-Enhanced Raman Scattering Based on the Shift of Characteristic Peak
Si-Qi YUE ; Zhan-Hao MO ; Jun-Qi ZHAO ; Xin QI ; Ling JIN ; Can-Can CUI ; Cheng-Yan HE ; Bing ZHAO
Chinese Journal of Analytical Chemistry 2024;52(2):231-239,中插11-中插13
Acute cerebral infarction(ACI)has the characteristics of onset nasty and high mortality,and thus the rapid determination of the occurrence and development of ACI plays a key role in the diagnosis,treatment and prognosis of ACI patients.It has shown that the serum level of human haptoglobin(Hp)is related to ACI.In this study,surface enhanced Raman scattering(SERS)combined with immune recognition was applied to establish a quantitative analysis method for serum Hp.Firstly,the SERS substrate of silver nanoparticles was prepared on silicon wafer,and 4-mercaptobenzoic Acid(MBA)was used as a Raman probe by forming Ag—S bond and connecting it on the surface of nanoparticles.The carboxyl group of MBA was linked to amino group of self-made high-affinity antibody through forming CO—NH structure thus forming a SERS self-assembled chip of Hp(Ag/MBA/anti-Hp).Hp in serum could be specifically captured by antibodies on SERS substrate,which caused the shift of SERS characteristic peak of MBA.The results showed that there was a good linear relationship between the logarithm of Hp concentration and the SERS characteristic peak shift of MBA.The detection range was 1-1000 ng/mL(R2=0.988).The Hp concentrations in serum of 90 ACI patients were determined by this method,and the results were consistent with those of ELISA method,which proved the practicability and accuracy of this method.This method was highly specific,simple and convenient,which could realize the specific recognition and quantitative analysis of serum Hp,so as to be an effective means for clinical detection of serum Hp,thus providing a reference for the treatment and prognosis of ACI.
4.Efficacy and safety of remimazolam tosylate for general anesthesia in morbidly obese patients
Gong CHEN ; Yan-Xi LU ; Jin LI ; Fan ZHANG ; Can-Can CHENG ; Xin-Lin YIN ; Sai-Ying WANG ; Huan CHANG
Chinese Pharmacological Bulletin 2024;40(5):859-864
Aim To evaluate the effectiveness and safety of remimazolam tosylate for administering general anesthesia in morbidly obese patients.Methods This clinical trial was conducted at a single center from De-cember 2021 to October 2023.It assessed 108 morbid-ly obese patients(body mass index,BMI≥40)who underwent laparoscopic sleeve gastrectomy.Patients were randomly assigned to either the remimazaolam group(Group R)or the propofol group(Group P)for general anesthesia induction and maintenance.The primary outcome was to compare the incidence of ad-verse events and postoperative recovery characteristics between the two groups.Results During induction pe-riod,the incidence of adverse events was higher in group P,including hypotension(P<0.01),hypox-emia(P<0.05),bradycardia(P<0.01),and in-creased vasopressor requirement(P<0.05).The time to loss of consciousness and BIS falling to 60 was shor-ter in group P than in group R(P<0.01).There were no statistically significant differences between the two groups in terms of postoperative quality of recovery(QoR-40 score),24-hour postoperative pain visual an-alogue scale(VAS)scores and morphine consump-tion.In conclusion,remimazolam tosylate,utilized for anesthesia induction in morbidly obese patients,signif-icantly reduced hypotension and hypoxemia compared to propofol,while it could also maintain similar postop-erative recovery quality.Conclusions Remimazolam is effective in reducing the incidence of hypotension and hypoxaemia during the induction period of general anaesthesia in morbidly obese patients and it is compa-rable to propofol in terms of quality of postoperative re-covery.
5.Isolation of a HBV-PreS2 epitope from a random peptide library displayed on the bacterial flagellin.
Zhong-tao XIN ; Yan-ning XUE ; Ya-ping GAO ; Can-quan MAO ; Chuan LIU
Acta Academiae Medicinae Sinicae 2003;25(1):56-59
OBJECTIVETo establish an improved procedure for isolation and identification of epitopes from a random peptide library displayed on the bacterial surface.
METHODSEpitopes were screened from FliTrx random peptide library by a monoclonal antibody 3B9 against HBV-PreS2 protein. The enrichment was monitored in each round. Higher affinity clones were obtained by increasing the washing strength and randomly selected for sequencing and Western blot analysis.
RESULTSClones specifically binding to antibody were enriched in each round. Ten sequences were obtained from sixteen sequenced clones, seven of them contained the common motif RXRGXY with high homogeneity to 135-140 amio acids in HBV-PreS protein and have positive results in Western blot analysis. The other three sequences have no typical motif RXRGXY and showed different Western blot results.
CONCLUSIONSIt's easy and quick to drive epitopes from a random peptide library displayed on the bacterial surface.
Amino Acid Sequence ; Animals ; Antigens, Viral ; immunology ; Bacterial Proteins ; immunology ; Flagellin ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; isolation & purification ; Immunodominant Epitopes ; immunology ; Molecular Sequence Data ; Peptide Library ; Protein Precursors ; genetics ; immunology ; isolation & purification
6.Construction of Saccharomyces cerevisiae Mutant Deficient in adh2 and ald6 Genes
Yan-Zun WANG ; Juan-Juan LEI ; Xian-Zhang JIANG ; Yuan-Yuan GAO ; Xin LI ; Can-Hua LAN ; You-Qiang CHEN ; Ru-Kai CHEN ; Jian-Zhong HUANG
Microbiology 1992;0(02):-
The purpose of this investigation is to improve ethanol production and decrease acetate formation in Saccharomyces cerevisiae strain YS2-?adh2.The strain YS2-?adh2 with deleted alcohol dehydrogenase Ⅱ(adh2) gene was isolated in our lab with higher ethanol production than that of the strain YS2.The ace-taldehyde dehydrogenase Ⅵ(ald6) gene encoded a cytosolic acetaldehyde dehydrogenase,a key enzyme of the pyruvate dehydrogenase(PDH) bypass,transfers acetaldehyde to acetate.To disrupt ald6 gene of the strain YS2-?adh2,ald6 gene targeting cassettes were synthesized by long flanking homology PCR(LFH-PCR) and then were transformed into YS2-?adh2 mutants by LiAc/SS Carrier DNA/PEG method.Positive transformants were selected with G418 and further confirmed by PCR.Once correctly integrated into the genome,the selective marker was rescued by transforming the plasmid pSH65 into the positive transformants and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.We named the ald6 gene knocked-out strain as YS2-?adh2-?ald6 which has a 12.5% higher ethanol production and a 18% lower acetate formation compared to the strain YS2.
7.Study on monitoring and early warning index system for schistosomiasis in Poyang Lake eco-economic region
Wen-yan FAN ; Xin-han ZHAO ; Jin-can WU ; Shao-xin HUANG ; Xiao-bo WANG ; Xin WANG
Chinese Journal of Disease Control & Prevention 2019;23(4):421-425
Objective To establish a scientific, operational monitoring and early warning index system for schistosomiasis, so as to provide scientific evidence for promoting the scientification and standardization of early warning system in Poyang Lake Eco-economic Region. Methods Two rounds of Delphi experts’ interviews were applied to construct Index system. The weight value of each indicator was determined by the Analytic Hierarchy process and improvable proportionate allocation method. Reliability, validity of index system and the rationality of index weight distribution can be evaluated in site investigation. Results The Index system included 3 first-order indicators, 9 second-order indicators, and 35 third-order indicators. The 3 first-order indicators were endemic status, environmental and social factors, control measures, with the weight value of 0.531 0, 0.101 5 and 0.367 5, respectively. For the 9 second-order indicators, the highest weight value was for Infection status of human and livestock (0.179 5)and the lowest for social factors(0.050 6). During site investigation, the Cronbach’s alpha and spit half reliability of the total index system and three first-order indicators were all over 0.90, the Kendall W coefficient for the data collected in site investigation and Delphi consultation was 0.742 (P=0.018). Conclusions The Monitoring and Early Warning Index System for Schistosomiasis is suitable for the infection status of Poyang Lake Eco-economic region. The reliability and validity of index system are satisfactory, and the indicator weight distribution is rational.
8.Analysis of post-thaw infused cell dose for predicting engraftment after unrelated cord blood transplantation.
Jie-Ying WU ; Can LIAO ; Jin-Song CHEN ; Zun-Peng XU ; Yan LI ; Xin SUN ; Shao-Qing WU ; Xue-Wei TANG ; Yan LU ; Gui-E XIE
Journal of Experimental Hematology 2011;19(3):754-758
This study was purposed to investigate the role of post-thaw infused donor cells for predicting engraftment and hematopoietic reconstitution after unrelated cord blood transplantation (UCBT). The retrospective analysis was performed on clinical data of 97 children with malignant or non-malignant diseases received single unit UCBT from August 1999 to April 2010. The impact of pre-freezing and post-thaw cell dose of total nucleated cells (TNC), CD34(+) cells and colony-forming units-granulocyte/macrophage (CFU-GM) on engraftment and hematological recovery after UCBT was analyzed. Unrelated donors were from Guangzhou cord blood bank (GZCBB) entirely. The results indicated that the pre-freezing TNC (/kg) (mean ± SD: 7.65 × 10⁷ ± 4.26 × 10⁷; median: 6.34 × 10⁷), CD34(+)cells (/kg) (mean ± SD: 4.64 × 10(5) ± 4.47 × 10⁵; median: 3.03 × 10⁵) and CFU-GM (/kg) (mean ± SD: 0.79 × 10⁵ ± 1.09 × 10⁵; median: 0.57 × 10⁵) showed a good correlation with their post-thaw counterparts including TNC(/kg) (mean ± SD: 6.98 × 10⁷ ± 4.12 × 10⁷; median: 6.00 × 10⁷), CD34(+)cells (/kg)(Mean ± SD: 6.86 × 10⁵ ± 8.56 × 10⁵; Median: 4.17 × 10⁵), and CFU-GM (/kg) (mean ± SD: 0.52 × 10⁵ ± 0.52 × 10⁵; median: 0.39 × 10⁵) (r = 0.952, p < 0.001; r = 0.794, p < 0.001; r = 0.478, p < 0.001). Either the pre-freezing or post-thaw number of infused CFU-GM was significant higher in patients who achieved engraftment (n = 70) than those who suffered graft failure (n = 22) (p = 0.023 and 0.011, respectively), but no significant difference of TNC and CD34(+) cells dose (pre-freezing or post-thaw) were found between these two groups. Pre-freezing CFU-GM, TNC, CD34(+) cell dose negatively correlated with the time of neutrophil engraftment (r = -0.285, p = 0.018; r = -0.396, p = 0.002; r = -0.373, p = 0.002), as well as the post-thaw number of TNC and CD34(+) cells (r = -0.260, p = 0.031; r = -0.483, p < 0.001), whereas only pre-freezing CD34(+) cells showed a significant correlation with platelet engraftment time (r = -0.352, p = 0.013). It is concluded that the CFU-GM amount is useful for predicting engraftment of UCBT, while pre-freezing hematopoietic cell doses show superior correlation with the speed of engraftment and hematopoietic reconstitution than their post-thaw counterparts in pediatric recipients, suggesting that it is essential to perform hematopoietic potency assay on each cord blood unit prior to listing or release for administration.
Adolescent
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Antigens, CD34
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blood
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Blood Banks
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Child
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Child, Preschool
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Cord Blood Stem Cell Transplantation
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methods
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Female
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Fetal Blood
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cytology
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Graft Survival
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Granulocyte-Macrophage Progenitor Cells
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Humans
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Infant
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Male
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Retrospective Studies
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Tissue Donors
9.Polymorphisms analysis of short tandem repeat loci D21S1433, D21S1442, D21S1444, D21S2051 in Guangdong Han nationality in China.
Xin YANG ; Can LIAO ; Yi-ning HUANG ; Yan LI ; Cui-xing YI ; Sao-qing WU ; Ming PAN ; Sun-yan HU ; Si-ming YUAN
Chinese Journal of Medical Genetics 2006;23(6):689-691
OBJECTIVETo investigate the polymorphic distribution of short tandem repeat (STR) sequences D21S1433, D21S1442, D21S1444, D21S2051 in Guangdong Han nationality in China.
METHODSUsing quantitative fluorescens PCR technology, the authors analyzed 200 unrelated samples to acknowledge the allele frequency, heterozygosity and other genetic information.
RESULTSD21S1433, D21S1442, D21S1444, D21S2051 were tested in 200 samples, which were tested to be statistical according to Hardy-Weinberg equilibrium (P> 0.05), 9, 10, 9 and 5 alleles were detected separately in each STRs. The heterozygosity of each STR was 0.818, 0.820, 0.770, and 0.261. The polymorphic information content > 0.7 in D21S1433, D21S1442, D21S1444, while D21S2051 owned only 0.247 polymorphic information.
CONCLUSIOND21S1433, D21S1442, D21S1444 are found to have high heterozygosity and polymorphic information content, and they could provide useful markers for genetic purposes, while D21S2051 is not informative in Guangdong Han nationality in China.
Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; Heterozygote ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Pregnancy ; Tandem Repeat Sequences ; genetics
10.HLA-B gene polymorphism detected by high-resolution sequence-based typing in Guangdong Han populations.
Yan LI ; Xue-wei TANG ; Jie-ying WU ; Si-min YUAN ; Xin YANG ; Yi-ning HUANG ; Can LIAO
Chinese Journal of Medical Genetics 2006;23(1):50-54
OBJECTIVETo investigate the genetic polymorphism of HLA-B locus in Guangdong Han population and compare the characteristic of the allele frequency distribution with that in other populations.
METHODSA total of 562 cord blood samples from Guangzhou Cord Blood Bank were analyzed by sequence-based typing. Then the sequences encompassing exons 2, 3, and 4 for HLA-B gene were analyzed by direct sequencing of PCR products. The allele frequency distribution of HLA-B in this population was compared with that in other populations.
RESULTSA total of 59 different HLA-B alleles were detected, and among them were 6 HLA-B alleles with frequencies higher than 5%: HLA-B*4601 (14.5%), HLA-B*400101 (14.4%), HLA-B*1502 (11.5%), HLA-B*1301 (8.6%), HLA-B*5801 (8.1%) and HLA-B*380201 (6.4%); the total frequency of these six alleles was 63.5%. At the same time, there were 30 kinds of HLA-B allele with frequencies lower than 0.5%; the total frequency of these alleles was 4.9%. Maximum variation at HLA-B was seen in the HLA-B*15 allele family (nine alleles). Comparison of the HLA-B frequencies in different populations showed a close relationship of Guandong Han population with the Chinese populations from Hong Kong and Singapore, respectively.
CONCLUSIONThe results have shown the characteristic of HLA-B distribution and provided more accurate genotypic data that may serve as normal reference value for the Han population in Guangdong, China.
Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Ethnic Groups ; Female ; Gene Frequency ; Genetics, Population ; HLA-B Antigens ; genetics ; Humans ; Male ; Polymorphism, Genetic