1.Hyphenation of C18 Gel Silica Column and Flow Injection withInductively Coupled Plasma-Mass Spectrometry andIts Application on the Seawater Analysis
Chaoyong YANG ; Xihong WU ; Shen GU ; Zhixia ZHUANG ; Xiaoru WANG
Chinese Journal of Analytical Chemistry 2001;29(3):283-286
An online enrichment-separation system of C18 and inductively coupled plasma-mass spectrometry (ICP/MS) with flow injection (C18-FI-ICP/MS) was established for the determination of trace heavy metals in seawater. The optimized experimental parameters including pH,C18 column,concentration of methol in elute and the velocity of elute were investigated. This system was applied to the online preconcentration , separation and determination of trace heave metals in seawater samples successfully. The detection limit (3σ) for Cd, Pb, Co, Ni and Zn was 0.03, 0.09, 0.1, 0.1, 0.3 μg/L respectively. The RSD of Cd, Pb, Co, Ni, and Zn for six measurements of seawater was 6.8%,3.4%,1.3%,2.6%,0.5%,respectively. The accuracy was verified with spiked recovery test. The recovery for five elements was 91.3%,95.1%,100.4%,100.3%,95.2%, respectively.
2.Preparation, Characterization and Application for Genomic DNA Extraction of a Magnetic Microsphere with Surface Charge Switching
Huabin HUANG ; Zhixia ZHUANG ; Chaoyong YANG ; Xiaoru WANG
Chinese Journal of Analytical Chemistry 2014;(11):1598-1603
SuperparamagneticSiO2@Fe3O4microspherewithcore-shellstructurewaspreparedbythe method of hydro-thermal and St?ber method, and modified by (3-aminopropyl) triethoxysilane ( APTES) . The obtained amino-Fe3 O4@SiO2 microsphere with controlled surface charge properties was characterized by SEM and TEM, and the surface electric property of amino-microsphere was investigated by Zeta potential measurement. The microsphere was used to extract DNA from human blood, and the solid phase extraction method by microspheres was developed. The mechanism of action between Aminated-Fe3 O4@SiO2 microsphere and DNA was explored, and the gel electrophoresis and PCR test were done on the extraction product. The result showed that genomic DNA with high purity was successfully extracted from whole blood by using amino-Fe3 O4@SiO2 microsphere, with the extraction rate about 70%, the saturated adsorption capacity for DNA was about 40 ng/μg, and the elution could be directly used for further bio-analysis.
3.Characterization of Cd and Cu Complexing with Metallothionein by Eletrospray Ionization Mass Spectrometry
Jincan SHEN ; Weixiang YE ; Haining KANG ; Liya GE ; Zhixia ZHUANG ; Xiaoru WANG
Chinese Journal of Analytical Chemistry 2009;37(7):975-979
Mammal metallothionein(MT) folds into two separate domains that exhibit different structure and metal binding propertity independently, the study of the strategy of metal ions binding with MT would give better understanding of their exact biological functional mechanisms. In this study, a method using eletrospray ionization mass spectrometry (ESI-MS) phase liquid chromatography and identified by ESI-MS. Different amounts of Cd or Cu were then added in MT-2a samples and ESI-MS was employed to determine the mass difference of MT in different samples. The results Cd2+4S11; while Cd is attached in separate binding sites for the formation of Cd2+3S9 cluster, which intermediately formed with five and six Cd ions were detected. For the Cuprous ions, it prefers to cooperatively bind in β-domain with the form of Cu4-MTβ. The binding form in β-domain would convert from Cu4 into more Cu binding form with the addition of Cu. When high concentration of Cu was added in samples, the result suggested that
4.Application of Electrical Driven and Cationic Exchange Membrane to Capture and Enrich Deoxyribonucleic Acid
Huabin HUANG ; Zhixia ZHUANG ; Wenhua ZHANG ; Jia HU ; Chaoyong YANG ; Xiaoru WANG
Chinese Journal of Analytical Chemistry 2010;38(1):143-146
Based on the selective permeability of cationic exchange membrane and the characteristic of electri cal driven, a new and non-damage apparatus that can be used to separate compounds with different electric property was built to capture and enrich DNA.λ-DNA, as a model compound, was used to study the charac teristic of capturing and enriching DNA in this device.Factors that might affect the efficiency such as voltage and flow speed were optimized and gel electrophoresis was done to study if DNA was denaturation, PCR was done to prove the concentration can be used to further bioanalysis.Results were got; the apparatus could used to capture and enrich DNA scathelessly under the condition of low-voltage, the concentrate multiple was 62, and the recovery efficiency was accessible to 47%, concentration can be used for the next bioanalysis.Auto matic, easy to handle and little organic reagent using made it be advantageous, it could be miniatured to com bine with other biochips.This method had a large application space in field of DNA extraction, purification and concentration.
5.Epidemiological and clinical characteristics of 323 chronic hepatitis C patients
Zhixia DONG ; Qing XIE ; Xiaogang XIANG ; Hui WANG ; Huijuan ZHOU ; Yan ZHUANG ; Yumin XU
Chinese Journal of Infectious Diseases 2010;28(4):218-221
Objective To analyze epidemiological and clinical characteristics of chronic hepatitis C (CHC) patients. Methods The clinical data of 323 CHC patients were collected. The transmission modes, clinical manifestations and virological features were recorded. The liver histological change was also analyzed in 39 cases whose liver biopsy samples were available. The comparison between two groups was performed by t test. Results Among the 323 CHC patients, 135 cases (41. 8%) had history of blood or blood products transfusion. Sixty-seven cases (20. 7%) had undergone surgery and trauma operation. Nineteen cases (5. 9%) had history of Chinese medicine acupuncture. Eighteen cases (5.6%) had undergone hemodialysis. Two patients (0.6%) were infected through vertical transmission. Twenty-one cases (6. 5%) had history of intravenous drug use and two cases (0. 6%) had history of unsafe sexual contact. The possible transmission routes for the other 78 cases (24.1%)were unknown. Fourteen patients (4. 3%) were co-infected with hepatitis B virus (HBV). The major prevalent genotypes were hepatitis C virus ( HCV) genotype lb and 2a, which were 145 cases (65. 3%) and 21 cases (9. 5%) respectively. HCV viral loads were as high as 1 × 105 IU/mL in 74 cases (26. 1%) and 1× 106 IU/mL in 103 cases (36. 4%). Twenty-three patients (7.1%) developed obvious clinical manifestations. Among 39 patients undergoing liver biopsy, 14 cases (35. 9%) had hepatic inflammation activity index (HAI)≥4, six cases (15. 4%) had fibrosis stage (F) ≥3, four cases (10. 3%) had HAI≥4 and F≥3. Conclusions The most common HCV transmission modes are blood transfusion and use of blood products. However, surgery and trauma operation should be paid more attention. Besides blood transfusion, the transmission modes of intravenous drug injection, hemodialysis and traditional Chinese medicine acupuncture are increasing. The major HCV genotypes are lb and 2a. The viral loads of most patients are relatively high. Most patients infected with HCV don't show any obvious hepatitis symptoms and physical signs. However, the liver biopsy results from 39 patients suggest that most patients develop liver histological changes.
6.Liver histological changes and influencing factors in patients with chronic hepatitis C
Zhixia DONG ; Huijuan ZHOU ; Xiaogang XIANG ; Yan ZHUANG ; Hui WANG ; Simin GUO ; Honglian GUI ; Weiliang TANG ; Gangde ZHAO ; Qing XIE
Chinese Journal of Infectious Diseases 2011;29(9):534-539
ObjectiveTo investigate the histological features as well as the factors influencing liver disease progression in Chinese patients with chronic hepatitis C (CHC). MethodsA total of 102 CHC patients who underwent percutaneous liver biopsy between August 2007 and May 2010 were recruited. Age, gender, body mass index (BMI) and transmission route of recruited patients were recorded. Serum levels of alanine transaminase (ALT) and aspartate transaminase (AST), HCV genotypes, HCV viral load and liver histological changes were detected. Statistical analysis was done by t test and Logistic regression. ResultsThe serum levels of ALT and AST in CHC patients with histological activity index (HAI) ≥4 were much higher, while platelet (PLT) counts were lower than those with HAI <4(t=2.209, 2. 298 and 2. 565, respectively; all P<0.05). Likewise, in patients with F≥3, the serum levels of ALT and AST as well as the mean age and the duration of infection were significantly elevated compared with F < 3 group ( t = 3.497, 2. 758, 2. 340 and 2. 570,respectively; all P<0. 05), while PLT counts were much lower (t = 2. 761, P=0. 007). The unvariate predictors for HAI≥4 were female, ALT>1 × upper limits of normal (ULN), AST level,F≥3, HCV RNA≥6 lgIU/mL and PLT counts. By mutivariate analysis, the Ishak stage score was the only independent predictor for HAI≥4 (OR 3.098, 95%CI 1.884-5. 092; P<0.01). Finally,the univariate predictors for F≥3 were age, BMI≥24 kg/m2 , ALT>1 × ULN, AST level, HAI≥4,PLT counts and duration of infection≥ 15 years. Multivariate analysis revealed that age (OR 1. 074,95%CI 1.006-1. 146; P=0.033), ALT level (OR 1. 035, 95%CI 1.015-1.055; P<0.01), ASTlevel (OR 0. 969, 95%CI 0. 948-0. 990; P=0. 005), the duration of infection ≥15 years (OR 37. 215, 95%CI 5. 816-238. 127; P<0.01) and HAI≥4 (OR 1. 939, 95%CI 1. 426-2. 636; P<0.01) were independent predictors for F≥ 3. ConclusionAge, ALT level, AST level, duration of infection≥15 years, HAI≥4 are independent predictors for liver fibrosis.
7.Differences of endoscopic features between undifferentiated-typed early gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma
Xiaowan WU ; Qian ZHUANG ; Jing WANG ; Dafan CHEN ; Zhixia DONG ; Yueqin QIAN ; Lungen LU ; Xinjian WAN ; Hui ZHOU
Chinese Journal of Digestive Endoscopy 2021;38(11):894-900
Objective:To analyze and compare the features of undifferentiated-typed early gastric cancer (UD-EGC) and gastric mucosa-associated lymphoid tissue(MALT) lymphoma under white light endoscopy (WLE) and magnifying endoscopy-narrow band imaging (ME-NBI).Methods:Data of patients with complete endoscopic images of WLE and ME-NBI in Shanghai General Hospital, Shanghai Jiao Tong University from March 2015 to July 2019 were retrospectively analyzed.Twenty-six UD-EGC patients and seven gastric MALT lymphoma patients in ⅠE1 stage were included, and the characteristics of the two diseases under WLE and ME-NBI were compared and summarized.Results:There were no significant differences in age, sex or infiltration depth of lesions between the two groups.Under WLE, UD-EGC was often manifested as a single lesion located in the lower part of the stomach, with unclear lesion boundaries. While MALT lymphoma lesions were mostly multifocal with clear boundaries, located in the middle of the stomach. Under ME-NBI, the microsurface pattern of UD-EGC showed dilation or disappearance of areas between the recesses, and the spiral microvascular pattern. However, the microsurface pattern of MALT lymphomas were characterized by " cross-road traffic sign" , " pebble sign" , and the presentation of residual glandular duct at the lesion was similar to that of Helicobacter pylori ( HP)-related gastritis. Furthermore, the microvascular pattern of MALT lymphomas often showed " tree like appearance (TLA)" . After HP eradication therapy, the morphology of microsurface pattern and microvascular pattern in the original lesion area gradually returned to normal. Conclusion:UD-EGC and gastric MALT lymphoma showed particular features in the number, site and boundary under WLE, and they showed significantly different microsurface pattern and microvascular pattern under ME-NBI. Differentiation of the two diseases will help reduce the risk of missed diagnosis and misdiagnosis.
8. Characteristic Analysis and Hepatic Transcriptomics of High-fat Diet-induced Cholesterol Gallstone Model in Mice
Qian ZHUANG ; Zhixia DONG ; Xin YE ; Jinnian CHENG ; Xinjian WAN
Chinese Journal of Gastroenterology 2020;25(10):581-587
Background: The mouse model of high-fat diet-induced cholesterol gallstone is widely used in researches of pathogenesis, prevention and treatment of gallstones. Aims: To investigate the characteristics and hepatic transcriptomics of the mouse model of high-fat diet-induced cholesterol gallstone. Methods: Twenty male C57BL/6J mice were randomly allocated into chow diet (control) group and lithogenic diet (LD) group. After 8 weeks, the occurrence of gallstone was observed; the serum lipids and gallbladder bile lipids were detected; and the differentially expressed hepatic genes between the two groups were identified with Illumina NovaSeq sequencing systems. The enrichment analysis was mapped in GO and KEGG pathway databases. Real-time PCR was used to detect the expressions of genes related to bile acid synthesis in the liver. Results: The cholesterol gallstone formation rate was 100% in LD group, whereas no gallstone was observed in control group. Hepatomegaly and steatosis were obvious in mice of LD group. The serum levels of total cholesterol and triacylglycerol, as well as the cholesterol content and cholesterol saturation index of the gallbladder bile in LD group were significantly higher than those in control group (P<0.05). A total of 1 330 differentially expressed genes were identified by high-throughput sequencing and bioinformatics analysis. KEGG analysis showed that the differentially expressed genes were mainly enriched in lipid metabolism, bile secretion, and insulin secretion pathways. GO analysis showed that fatty acid metabolic process-related pathways were significantly enriched. Both hepatic transcriptomics analysis and real-time PCR showed that the expression levels of genes related to bile acid synthesis, including CYP7A1, CYP8B1 and CYP7B1 decreased significantly in the liver of LD group as compared with those of control group (P<0.05). Conclusions: The metabolism of cholesterol, bile acids and fatty acids is significantly disordered in mice with cholesterol gallstone. Transcriptomics analysis can screen out the differentially expressed genes that play roles in the formation of cholesterol gallstone, so as to provide references for studies focusing on these topics.