1.Mutation analysis of STK11 gene in patients with Peutz-Jeghers syndrome
Changyuan WANG ; Hua LIU ; Jinbao ZONG ; Shiguo LIU ; Tongxin SHI
Chinese Journal of Dermatology 2014;47(1):42-44
Objective To study the mutation of STK11 gene in a Chinese family and a sporadic patient with Peutz-Jeghers syndrome (PJS),and to provide a basis for genetic diagnosis and counseling.Methods One sporadic patient and two patients from a family with PJS were collected,all of whom had typical mucosal pigmentation and gastrointestinal polyposis.Blood samples were obtained from the two patients and six unaffected relatives in this family,the sporadic patient,and 100 healthy controls.DNA was extracted,and PCR was performed to amplify nine exons and their adjacent introns in the STK11 gene followed by direct sequencing.The sequencing results were aligned to the published sequence of STK11 gene from Genbank.Results No mutation was found in the STK11 gene of any of the patients,unaffected relatives,or healthy controls.Conclusions Genetic heterogeneity exists in Peutz-Jeghers syndrome,hinting that there may be other causative genes or sites for this entity.
2.RESEARCH PROGRESS ON ENZYMATIC HYDROLYSIS OF NITRILE
Wen-Yong LOU ; Min-Hua ZONG ; Sen-Lin LIU ;
Microbiology 1992;0(06):-
Enzymatic hydrolysis of nitrile was performed under mild conditions and afford high efficiency and selectivity,thus being highly potential for the synthesis of optically active carboxylic acids and their derivatives.The type of nitrile-hydrolyzing enzymes and hydrolysis reactions,reaction characteristics,factors influencing the hydrolysis and the prospect for its application to industrial production were reviewed in this paper.
3.Variation and Functional Study of Q Promotor of Epstein-Barr Virus in Nasopharyngeal Carcinoma
Yufan HUANG ; Yiji LIAO ; Wenju LIU ; Wenfeng HUA ; Haixia DENG ; Shijuan MAI ; Dan XIE ; Yongsheng ZONG
Journal of Sun Yat-sen University(Medical Sciences) 2010;31(1):23-27
[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.
4.Contents Determination of Citric Acid in Fentanyl Citrate Raw Materials and Its Injection by Ion Chroma-tography
Xun MA ; Panpan LI ; Guixia LIU ; Yanping ZONG ; Wei SUN ; Guangzhi SHAN ; Hua CHEN ; Nan NAN
China Pharmacy 2017;28(18):2536-2538
OBJECTIVE:To determine the contents of citric acid in fentanyl citrate raw materials and its injection by ion chro-matography. METHODS:The determination was performed on Thermo Dionex IonPacTM AS11-HC column with mobile phase con-sisted of potassium hydroxide (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and sample size was 20 μL. The detector was suppressed conductivity detector. RESULTS:The linear range of citric acid were 0.1157-74.05 μg/mL(r=0.9995). The limit of quantitation was 0.1150 μg/mL,and the limit of detection was 0.0400 μg/mL;RSDs of preci-sion,stability and reproducibility tests were all lower than 2.0%;the average recoveries were 99.6%-101.5%(RSD=0.68%,n=9). CONCLUSIONS:The method is environmentally-friendly and simple with good accuracy and precision,and suitable for the contents determination of citric acid in fentanyl citrate raw materials and injection.
5.Evaluation of Clinical Detection Methods for Detection of Helicobacter Pylori Infection in Children
zong-ping, JIN ; yu-hua, LIU ; li, LI ; ai-jun, YANG
Journal of Applied Clinical Pediatrics 2006;0(19):-
Objective To explore the optimal methods to detect helicobacter pylori(Hp)antigen in children.Methods 13C labeled urea breath test(13C-UBT)was performed on 937 children of alimentary department from Sep.2000 to Feb.2006.Gastric mucosa biopsy of 96 children was detected.Hp stool antigen(HpSA)status of the 557 children were evaluated.Both 13C-UBT and HpSA were assayed in 105 children from Apr.2003 to Apr.2004.Method of 13C-UBT was taken as the golden standard on diagnosis of Hp infection.Results Forty-one point seven percent children was positive for 13C-UBT.Forty point six percent children was positive for gastric mucosa biopsy.Thirty-eight point two percent was positive for HpSA.The difference was not significant.2.Among the 105 children performed by both 13C-UBT and HpSA,41.9% was positive for 13C-UBT and 39.0% for HpSA.Taking 13C-UBT as the golden standard,sensitivity of HpSA to diagnose Hp infection was 91.8%,and its specificity was 81.8%.Consistency Kappa coefficient was 0.743(P=0),which denoted that there was no significant difference on the positive detection rate between 13C-UBT and HpSA(P=0.388).Conclusions 13C-UBT and HpSA as non-invasive technique is effective to detect the Hp antigen.Compared to 13C-UBT,as a convenient,noninvasive,economical method,HpSA detection is much more acceptable to children and their patients.
6.Effects of Mycobacterium vaccae vaccine on immunological function in elderly patients with stable chronic obstructive pulmonary disease
Zong-xiao SHANGGUAN ; Ren-guo ZHAO ; Ling-yun LIU ; Feng GONG ; Xiao CI ; Ming-hua ZHANG ;
Chinese Journal of Geriatrics 2010;29(5):382-385
Objective To investigate the effects of Mycobacterium vaccae vaccine on immunological function and clinical character in elderly patients with stable chronic obstructive pulmonary disease (COPD). MethodsA total of 100 elderly patients with stable COPD were randomly divided into immunotherapy group (group A, n= 50) and non-immunotherapy group (group B, n= 50), and normal control group (group C, n = 50). The levels of peripheral blood T-lymphocyte subsets (CD3+ , CD4+, CD8+ , CD4+/CD8+ ratio), natural killer cells (NK cells), immunoglobulins (IgG, IgA, IgM) and cytokines (IL-6, IL-8, TNF-a) were measured respectively before and after therapy. Group A and B were followed up for 1 year, then the times of acute outbreak and hospitalization of patients in the two groups were also compared. Results The levels of CD4 + ,CD4+/CD8+ ratio and NK cells in group A, B were significantly lower before therapy (P<0. 05~0. 01=, and the levels of IL-6, IL -8, TNF-a and IgA were significantly higher than in group C (P<0. 01=. After treatment with Mycobacterium vaccae vaccine in group A, the levels of CD4+ , CD4+/CD8+ ratio and NK cells were significantly higher (P<0. 05-0. 01= and IL-6, IL-8, TNF-a and IgA were significantly lower than before treatment (all P<0. 01=. These levels showed no significant changes in group B after treatment (P>0. 05). After 1-year follow-up, the times of acute outbreak and hospitalization on patients were statistically lower in group A than in group B (P< 0. 01 ).ConclusionsMycobacterium vaccae vaccine can improve cellular immunity function and reduce the times of acute outbreak and hospitalization in patients with stable COPD, so it has a higher clinical application value.
8.Correlation analysis between meteorological factors, biomass, and active components of Salvia miltiorrhiza in different climatic zones.
Chen-lu ZHANG ; Zong-suo LIANG ; Hong-bo GUO ; Jing-ling LIU ; Yan LIU ; Feng-hua LIU ; Lang-zhu WEI
China Journal of Chinese Materia Medica 2015;40(4):607-613
In this study, the growth and accumulation of active components of Salvia miltiorrhiza in twenty two experimental sites which crossing through three typical climate zones. The S. miltiorrhiza seedlings with the same genotype were planted in each site in spring, which were cultivated in fields with uniform management during their growing seasons till to harvest. The diterpene ketones (dihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone II(A)) in S. miltiorrhiza root samples were determined by using high-performance liquid chromatography (HPLC) method. The biomass of root (root length, number of root branches, root width and dry weight) was also measured. The results showed that tanshinone II(A) in all samples of each site were higher than the standards required by China Pharmacopoeia. It has been found there is a relationship between root shape and climate change. The correlation analysis between active components and meteorological factors showed that the accumulation of tanshinones were effected by such meteorological factors as average relative humidity from April to October > average vapor pressure from April to October > average temperature difference day and night from April to October > annual average temperature and so on. The correlation analysis between root biomass and meteorological factors exhibited that root shape and accumulation of dry matter were affected by those factors, such as average annual aboveground (0-20 cm) temperature from April to October > annual average temperature > average vapor pressure from April to October > annual active accumulated temperature > annual average temperature > average vapor pressure from April to October. The accumulation of tanshinones and biomass was increased with the decrease of latitude. At the same time, the dry matter and diameter of root decreased if altitude rises. In addition, S. miltiorrhiza required sunlight is not sophisticated, when compared with humid and temperature. To sum up, S. miltiorrhiza can adapt to a variety of climatic conditions and the southern warm humid climate is more conducive to its growth and accumulation of active components.
Biomass
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China
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Climate Change
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Drugs, Chinese Herbal
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analysis
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Ecosystem
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Plant Roots
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chemistry
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growth & development
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Salvia miltiorrhiza
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chemistry
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growth & development
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Temperature
9.Spaceflight-induced variation on biological traits and effective components of Cassia obtusifolia.
Ren-jun MAO ; Zhi-hong QI ; Rui-lian HAN ; Feng-hua LIU ; Yan LIU ; Zong-suo LIANG
China Journal of Chinese Materia Medica 2015;40(13):2571-2575
The dry seeds of Cassia obtusifolia were carried by the "ShenZhou 8" satellite and sowed after landing. Based on our pri- or study on SP1, the characteristics of plants growth, physiological index and content of effective components were examined. The results showed that the QC10, QC29 strains matured 5 d earlier compared with control. The plant height, across diameter and ground diameter of QC10, QC29, QC46 strains was superior to the control at whole growth period. The branch number increased ranging from 4 to 11 and the number of pods reached 321, 313,281, respectively, which was dramatically higher than the control (246). The yield of QC10, QC29, QC46 strains increased noticeably from 31.4 to 63.2 g. The 1000-seed-weight of QC10, QC29, QC46 strains was 25.86, 25.88, 24.06 g, while the control was 23.69 g. Compared to the control, the mass fraction of chlorophyll was enhanced 1.098, 1.016, 0.297 mg. There was no significant difference in aurantio-obtusin and chrysophanol content of seeds. Through two years research, three high-yield mutant strains were obtained. This study indicates that spaceflight-induced mutants could provide new germplasm for C. obtusifolia breeding and offers the theoretical basis for further utilization of spaceflight-induced mutation to breed high-quality C. obtusifolia strains.
Cassia
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chemistry
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genetics
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growth & development
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Mutation
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Space Flight
10.A new C21 steroidal saponins from Periplocae Cortex.
Ying LIU ; Yue OUYANG ; Zong-quan WANG ; Li QIAO ; Song LI ; Shao-hua ZHAO ; Min-yan LIU
China Journal of Chinese Materia Medica 2015;40(3):455-457
To study the chemical constituents of Periplocae Cortex, the separation and purification of 70% alcohol extract were carried out by column chromatographies on AB-8 macroporous resin, silica gel and preparative HPLC. The structure of the compounds were identified by NMR and TOF-MS. A new compound was isolated and identified as 21-O-methyl-Δ5-pregnene-3β, 14β, 17β, 21-tetraol-20-one-3-O-β-D-oleandropyranosyl(1-->4)-β-D-cymaropyranosyl-(1-->4)-β-D-cymaropyranosyl (1), named as periplocoside P.
Glycosides
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chemistry
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isolation & purification
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Periploca
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chemistry
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Pregnenes
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chemistry
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isolation & purification
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Saponins
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chemistry
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isolation & purification