2.Determination of 27 elements in Maca nationality's medicine by microwave digestion ICP-MS.
Gui-fang YU ; Hai-jie ZHONG ; Jun-hua HU ; Jing WANG ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(23):4545-4551
An analysis method has been established to test 27 elements (Li, Be, B, Mg, Al, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Sr, Mo, Cd, Sn, Sb, Ba, La, Hg, Pb, Bi) in Maca nationality's medicine with microwave digestion-ICP-MS. Sample solutions were analyzed by ICP-MS after microwave digestion, and the contents of elements were calculated according to their calibration curves, and internal standard method was adopted to reduce matrix effect and other interference effects. The experimental results showed that the linear relations of all the elements were very good; the correlation coefficient (r) was 0.9994-1.0000 (Hg was 0.9982) ; the limits of detection were 0.003-2.662 microg x L(-1); the relative standard deviations for all elements of reproducibility were lower than 5% (except the individual elements); the recovery rate were 78.5%-123.7% with RSD lower than 5% ( except the individual elements). The analytical results of standard material showed acceptable agreement with the certified values. This method was applicable to determinate the contents of multi-elements in Maca which had a high sensitivity, good specificity and good repeatability, and provide basis for the quality control of Maca.
Lepidium
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chemistry
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Mass Spectrometry
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methods
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Microwaves
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Reproducibility of Results
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Trace Elements
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chemistry
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isolation & purification
3.Screening, Identification of a Chitinolytic Bacterium Strain SWCH-6 and Its Optimal Chitinase Fermentation Conditions
Hai-Dong WANG ; Biao CHEN ; Jing-Sheng LUN ; Cheng WANG ; Zhong HU ;
Microbiology 1992;0(05):-
A bacterium, having high chitinolytic activity, was isolated from sediment of Shantou Bay, named SWCH-6. According to its physiological and biochemical characteristics and 16S rDNA sequence, it was identified as Aeromonas hydrophlilla. The optimal chitinase fermentation conditions of strain SWCH-6 were conformed by single-factor experiments and orthogonal experiments, they were colloidal chitin 25.0 g/L, tryptone 10.0 g/L, seawater 1 L, pH 8.5, 32 ℃ , 150 r/min for 72 h. In these conditions, its chitinase activity reached 0.39 U/mL. In addition, at 40℃ and pH 5.0, its chitinase performed the highest catalytic activity and its chitinase activity could be enhanced by Cu2+, Fe3+ and surfactant toween-80; weakened by Zn2+, Mn2+ and surfactants SDS, detergent powder, and there were some differences from orther chitinases.
4.Identification of novel genetic markers in Mycobacterium tuberculosis Beijing genotype strains
Xin JIANG ; Feng GAO ; Yi WANG ; Wen-Hong ZHANG ; Zhong-Yi HU ; Hong-Hai WANG ;
Chinese Journal of Infectious Diseases 2007;0(09):-
Objective To identify a unique protein as a novel genetic marker for rapid molecular typing of Mycobacteriutn tuberculosis Beijing genotype strains by comparing the proteome of Beijing genotype strains with non-Beijing strains.Methods Fifty-six clinical isolates of Mycobacterium tuber- culosis were analyzed by spoligotyping to determine genotypes.The two-dimensional electrophoresis (2 DE)was used to compare the global protein patterns between Beijing genotype strains and non Bei jing strains.Differential expressed proteins were measured by matrix assisted laser desorption ioniza tion lime of flight mass spectrometry(MALDI-TOF-MS).The data obtained from peptide mass fingerprinting were compared in protein database.The genes encoding differential expressed proteins and their upstream were sequenced.Results Forty nine of the 56 isolates were Beijing genotype strains and 7 isolates were non-Beijing strains.A unique protein Rv0927c was identified,which is absent in Beijing genotype strains compared with 7 non Beijing strains and H37Rv.There were two characteristic mutations in Beijing genotype strains,a deletion of AGC at nucleotide position 421 of Rv0927c and a 127 G→A muta- tion in the upstream of Rv0927c.but not in non Beijing strains and H37Rv.Conclusion Characteris tic mutations of Rv0927c in Beijing genotype strains can be used as a novel genetic marker for rapid molecular typing of Mycobacteriuln tuberculosis Beijing genotype strains and non Beijing strains.
5.Epidemiological analysis of human brucellosis in Ningxia from 2004 to 2009
Xiao-jia, SONG ; Feng, JIN ; Xing-zhong, HU ; Tian-bo, MA ; Hai-hong, WANG ; Li, LI
Chinese Journal of Endemiology 2011;30(1):92-95
Objective To learn the prevalence and trends of Brucella disease in Ningxia, in order to provide scientific basis for effective control of the disease. Methods Data of Brucella cases reported through city network from 2004 to 2009 in Yinchuan city, Shizuishan city, Wuzhong city, Guyuan city and Zhongwei city of Ningxia were reviewed and retrospectively analyzed. The data included demographic characteristics, treatment conditions and medical history so on related information. Analytical indicators include reported incidence;with patients' gender, age, regional distribution, urban and rural distribution;become chronic and associated factors;distribution of the cases reporting unit and so on. Results From 2004 to 2009, Ningxia had reported 349 cases of Brucellosis, no deaths, the annual incidence rates reported were 0.017/10 million, 0.543/10 million, 0.151/101 (295/54);The proportion of 34- to 40-year-old age group was higher than other age groups(27.5%, 96/349);Occupational distribution of patients was mainly farmers and herdsmen(70.2% ,245/349), in regional distribution of the patients, the highest percentage was Wuzhong city(61.9%,216/349), followed by Yinchuan city(22.9%,80/349);The proportion of patients in rural areas(97.4% ,340/349) was higher than urban(2.6% ,9/349);the proportion of patients converted to chronic was 11.2% (39/349). With age, the chance of patients converted to chronic was in a decreasing trend(odds ratio was 0.966);cases reported by Centre for Disease Control and Prevention accounted for 74.8%(261/349), by hospital accounted for 25.2%(88/349). Conclusions The reported incidence of Brucellosis in Ningxia is in a rapid upward trend year by year, the patients is mainly young men, the rate of converted to chronic is higher and the ability of hospital in founding and reporting of the cases is weaker.Comprehensive measures should be taken to increase funding, strengthen monitoring, and continuously improve the level of awareness and diagnosis of medical personnel for further strengthen the prevention and control of Brucellosis.
6.Dosimetric study of three dimension therapy plans in patients with squamous cell carcinoma of tongue receiving postoperative intensity-modulated radiotherapy or conventional radiotherapy
Hai-sheng, HU ; Chao, YAN ; Hui-feng, SHI ; Zhong-he, WANG ; Ming, GUO
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(6):698-701
Objective To evaluate the dose distribution in clinical target volume (CTV) and organs-at-risk (OARs) in three dimension therapy plans in patients with squamous cell carcinoma of tongue receiving postoperative intensity-modulated radiotherapy (IMRT) or conventional radiotherapy (CRT) by dosimetric study. Methods Thirty-five patients with squamous cell carcinoma of tongue were divided into CRT group(n=17) and IMRT group(n=18). All patients underwent head-and-neck immobilization with a thermoplastic mask and planning CT scan, and target volume and OARs were contoured. Dose calculation and plan optimization were performed. All three dimension plans passed quality assurance before treatment. The dosimetry of therapy plans with IMRT or CRT in target volume and OARs dose distribution was compared by dose-volume histogram (DVH), conformity index (CI) and homogeneous index (HI). Results There were significant differences in D95 (isodose line to cover 95 percent target volume), CI, HI, minimum dose and maximum dose in CTV of therapy plans between patients with IMRT and CRT(P < 0.01), and there was no significant difference in mean dose of CTV(P > 0.05). The radiation dose on salivary glands (both parotid glands and contralateral submandibular gland) in patients with IMRT was significantly lower than that in patients with CRT(P < 0.01). Conclusion Compared with dose distribution of CRT plans, there are more advantages in improving dose distribution at the target volume and sparing salivary glands in IMRT therapy plans in patients with squamous cell carcinoma of tongue.
7.Clinical significance of anatomic exposure of recurrent laryngeal nerve in thyroid surgery
Hai LI ; Xueqing JIANG ; Ming JIANG ; Yuan ZHONG ; Yonggang YANG ; Zhiyong WU ; Bo HU
Journal of Endocrine Surgery 2009;3(6):387-389
Objective To investigate the anatomic feature and exposing methods of recurrent laryngeal nerve(RLN) in thyroid surgery. Methods The data of 327 cases of thyroid tumor were analyzed retrospectively. The anatomic feature, injury and prevention of RLN were discussed. Of all the cases, 293 RLNs (242 cases) were anatomically exposed. Results The temporary injury rates of RLN was 1.65% (4/242) in the exposed group and 8.23% (7/85) in the unexposed group with a persistent injury rate of 2.35%(2/85). The differences of temporary injury rates and persistent injury rates between the two groups were statistically significant (P<0.05). 67.23% (197/293) of the RLNs had two branches before entering larynx, 61.09%(179/293) of RLNs were posterior to the inferior thyroid artery and 31.39% (92/293) anterior to the inferior thyroid artery, 4.09% (12/293) passed through the bifurcation of inferior thyroid artery, and 4. 13% (10/293) were not related to the inferior thyroid artery. The incidence of non-recurrent laryngeal nerve (inferior laryngeal nerve) was 0.68% (2/293). Conclusions The anatomic relation of RLN is relatively complicated along its journey. The injury of RLN could be reduced with designed exposal during the thyroid surgery.
8.Effect of mouse macrophage metalloelastase gene transfer into murine CT-26 colon cancer ceils on orthotopic tumor growth and angiogenesis
Hai SHI ; Jian-Ming XU ; Nai-Zhong HU ; Xuelong WANG ; Qiao MEI ; Junjun BAO ;
Chinese Journal of Digestion 2001;0(11):-
Objective To determine the correlation between mouse maerophage metalloelastase (MME)and vascular endothelial growth factor(VEGF)expression involved in angiogenesis of colon cancer.Methods A eDNA fragment coding for domainsⅠandⅡof MME was transfected into murine CT-26 colon cancer cells that were MME deficient.The enzymatic activity of recombinant MME was confirmed by cleavage of native substrate in vitro.An orthotopic implantation model was established by using MME-transfected cells and control cells.Tumor samples were subjected to in situ hybridization (ISH)and immunohistochemical staining(IHC)to detect expressions of MME and VEGF.The microvessel counting was used to assess angiogenesis of murine colon tumors.Results It was demon- strated that the tumor growth was significantly inhibited in MME-transfected group compared with pcDNA3.1 transfected and nontransfected groups(P<0.001).It was also found that,compared with pcDNA3.1-transfected and nontransfected groups,the microvessel formation in MME transfected group was significantly reduced(P<0.001).The expression of VEGF mRNA and protein was significantly lower in MME-transfected group than those in the controls,as demonstrated by ISH(MME-transfected group versus pcDNAa.1-transfected group,P=0.028;and versus nontransfected group,P=0.003) and by IHC(MME-transfected group versus pcDNA3.1-transfected group,P=0.025;and versus non- transfected group,P=0.008).Conclusions The MME gene transfected into murine colon cancer cells can effectively suppress the growth of orthotopic tumors by inhibition of vaseularity.Both MME and VEGF gene expression is highly associated with the vascularity of tumors,which may depend on a hal- ance between MME and VEGF expression.
9.Relationship between genetic polymorphism of VEGF and risk factor to lung cancer
Jing LIANG ; Xiao-Lin LIU ; Dian-Shui SUN ; Hai-Rong LIU ; Wei HU ; Ai-Zhong QU ; Yan LI ;
China Oncology 2006;0(11):-
Background and purpose:Vascular endothelial growth factor(VEGF) is a potent angiogenic mediator and angiogenesis has important effects on tumor growth and metastasis.The present study was to investigate the relationship between genetic polymorphism of VEGF and heredity risk factor of lung cancer.Methods:VEGF genotypes were determined by PCR-RFLP method in 171 patients with lung cancer and 172 healthy controls.Software PHASE 1.0 was used to construct the haplotypes of every individual.Unconditional logistic regression model was used to analyze the statistical association of genotypes or haplotypes in the two groups adjusted by gender and age. Results:Individuals with at least one-2578A allele had a significantly decreased risk of lung cancer compared with those carrying-2578CC genotype.When the analyses were stratified by gender,the combined-2578 CA and AA genotype,were associated with a considerably reduced risk of lung cancer(P=0.001,OR=0.303,95%CI=0.15 3-0.601).The distribution of the two haplotypes(936C/-2578C and 936C/-2578A) among overall lung cancer cases was significantly different from that among the controls(P=0.016,0R=0.317,95%CI=0.124-0.809 and P=0.018,OR=0.547, 95%CI=0.331-0.903).When the cases were categorized by tumor histology,the distribution of C-C haplotype in the adenocarcinoma(AC) group was associated with a substantially lowered risk of AC(P=0.004,0R=0.237,95%CI=0.090- 0.627),compared with the reference haplotypes.Conclusion:VEGF polymorphism may be a critical risk for the genetic risk factor to lung cancer.
10.Ultrastructure observation of rhesus bone marrow mesenchymal stem cell after transplantation of cornea
chun-ling, WEI ; Xiao-mei, SUN ; Zhong-kun, YANG ; Jie-jie, DAI ; Hai, LIU ; Xiang, JI ; Zhu-lin, HU
Chinese Journal of Experimental Ophthalmology 2011;29(9):793-798
BackgroundThe quest to look for seed cells is a hot spot of cornea transplant research in solving the problem of the lack of donor. Bone marrow mesenchymal stem cells(BMSCs) have been successfully induced into retinal ganglion cells(RGCs) in vivo,but the successful induction of BMSCs into corneal endothelial cells has not been reported.Objective This experiment was to study the transplantation of BMSCs on corneal endothelial surface using the splitting Descemet's membrane. MethodsFour healthy adult rhesus monkeys were divided into the experimental group ( 3 monkeys) and control group ( 1 monkey). Mesenchymal stem cells (MSCs) were isolated from bone marrow by density gradient centrifugation combined with adhering means. The cultured cells were identified by flow cytometry and its ability to differentiate was determined by allowing them to differentiate into adipocytes in vitro and labeled by 5-bromodeoxyuridine ( BrdU ) for subsequent identification. Corneal grafts of 7 mm in size with tearing of the Descemet' s membrane were prepared in the experimental group and control group. After labeling by 5-bromodeoxyuridine( BrdU ) ,cultured cells were transplanted onto the endothelial surface of cornea grafts in the experimental group, but no cultured cells were seeded in the graft of the control group. The corneal grafts were then sutured in situ, and were removed 1,2 or 3 months after operation to examine the distribution and connection between transplanted cells and their morphologic changes under the electron microscope. Results High purity MSCs were harvested by density gradient centrifugation combined with adhering method. Cultured cells reached confluency after 12 to 16 days, presenting with a spindle shape and parallel or swirling arrangement. Flow cytometry analysis showed that 94.26% of cells were positive for CD29,7. 51% for CD34 and 4. 02% for CD45. Larger nuclei filled with plastosomes, golgiosomes and rough endoplasmic reticula were found on the graft under the transmission electron microscope( TEM ). After 3 weeks, MSCs were differentiated into adipocytes where Oil Red O staining resulted in an orange-red staining in the cytoplasm and blue staining in the nuclei. The transplanted cells attached loosely on the endothelial surface of the corneal graft and came in contact with each other in one month. The shape of the cells appeared as spindle-shaped and polygonal after 2 months and became tightly packed after 3 months. The positive cells retained the BrdU label and presented with brown nuclei. No endothelia cells grew in the cornea graft in the control group, with an absence of BrdU labeling. Conclusions Mesenchymal stem cells can be transplanted onto the corneal endothelial surface successfully and form a monolayer using the centrifugation method, and present with good survival and proliferation ability.