Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

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Stem cell research in hepatocellular carcinoma
SUN Chengyi, ZUO Shi
Front. Med.. 2008, 2 (1): 1-4.  
https://doi.org/10.1007/s11684-008-0001-6

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The traditional view that adult human liver tumors, mainly hepatocellular carcinoma (HCC), arise from mature cell types has been challenged in recent decades. The results of several studies suggest that HCC can be derived from liver stem cells. There are four levels of cells in the liver stem cell lineage: hepatocytes, hepatic stem cells/oval cells, bone marrow stem cells and hepato-pancreas stem cells. However, whether HCC is resulted from the differentiation block of stem cells and, moreover, which liver stem cell lineage is the source cell of hepatocarcinogenesis remain controversial. In this review, we focus on the current status of liver stem cell research and their roles in carcinogenesis of HCC, in order to explore new approaches for stem cell therapy of HCC.
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Trends in serum lipid levels of healthy Chinese in Shanghai from 1973 to 1999
CHEN Haozhu, ZHOU Jun, CHEN Bin, ZHUANG Hanzhong, JIN Xuejuan, HAN Qingqing
Front. Med.. 2008, 2 (1): 5-10.  
https://doi.org/10.1007/s11684-008-0002-5

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We assessed the secular trends in the serum lipid levels in Shanghai residents from 1973 to 1999. Data from four independent, cross-sectional, population-based surveys were used to analyze the trends in serum lipid levels from 1973 to 1999. A standard protocol was applied across each survey in at least three independent surveys which were separated a 10-year study period. A total of 10733 (1385, 3302, 2399, and 3647 subjects who had complete data on serum lipids in 1973–1974, 1982, 1983 and 1997–1999, respectively) Shanghai residents aged from newborn to 80 years old were enlisted at random. Serum lipid levels were measured in a single laboratory throughout the entire study period. A non-significant increase was noted from 1973–1974 to 1982–1983 in the serum lipid profiles; however, a significant increase was observed from 1982–1983 to 1997–1999 in most of the age groups of both sexes. Mean serum total cholesterol (TC) levels increased significantly (P < 0.05) over the entire study period (overall period covered: 1973–1999) in most age groups of both sexes. In males, the changes were from 7.3% to 23.2% and in females, from 9.0% to 21.6%, except in umbilical blood samples and in females over 60 years where insignificant results were observed. Similar trends were observed in the mean serum triglyceride (TG) and the mean serum low-density lipoprotein cholesterol (LDL-c) levels, both in females and males. The majority show an upward trend. As for TGs, the changes were from 27.5% to 82.7% in males and from 13.2% to 54.6% in females. As for LDL-c the changes were from 34.9% to 71.0% in males and from 31.9% to 63.6% in females. Mean serum high-density lipoprotein cholesterol (HDL-c) levels were decreased significantly from 1982–1983 to 1997–1999, with the changes from -13.2% to -38.6% in males and from -9.8% to -21.7% in females. The ratio of total cholesterol to high-density lipoprotein cholesterol (TC/HDL-c) increased dramatically during this period because of a concurrent decline in the levels of HDL-c and the increase in the levels of TC. The prevalence of hypercholesterolemia (To achieve the normal range of serum TC and TG in healthy people of Shanghai, we determined the cut-off point in the cumulative frequency curve as the 95% percentile of the data collected by the survey in 1973–1974. The results show that the 95% cut-off point for TC was 220 mg/dL, while for TG, it was 160 mg/dL.) increased significantly (P < 0.05). In males aged 20 to 40 years old, the prevalence of hypercholesterolemia was from 1.6% to 7.79%, and in ages> 40 it was from 5.5% to 11.57%. In females, it was 0 to 4.68% and 6.5% to 16.2%, respectively. In general, the trends in the levels of TC, TG and LDL-c were increasing and trends in the levels of HDL-c were decreasing in most age groups, especially, in the younger age group of Shanghai residents from 1980s to 1990s. These worrying findings are significant for its public health implications. Health education and health promotion are necessary to prevent the disease prevalence from rising in the Chinese population.
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Diagnostic evaluation of plasma aldosterone concentration to plasma renin activity ratio in primary aldosteronism
ZHANG Huilan, WANG Daowen
Front. Med.. 2008, 2 (1): 11-14.  
https://doi.org/10.1007/s11684-008-0003-4

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Using the plasma aldosterone concentration to plasma renin activity ratio (PAC/PRA ratio) as the screening test of choice for primary aldosteronism in hypertensive patients, we studied the clinical characteristics and the diagnostic value of PAC/PRA ratio in primary aldosteronism. The plasma aldosterone concentration (PAC) and plasma renin activity (PRA) levels were measured by radioimmunoassay in 902 hypertensive patients from out-patient clinics or hospitals. One hundred and twenty-six suspected primary aldosteronism patients whose PAC/PRA ratio was > 25 ng/dL/ng/mL/hr had a lamellar computed tomography (CT) scan in the adrenal gland and follow-up visits. The proportion of primary aldosteronism in hypertensive patients was 14% (126/902). There were 54 patients with unilateral or bilateral hyperplasia and 25 patients with adenoma according to the CT scan. 39% (49/126) of the patients with primary aldosteronism had hypokalemia. Twenty-five patients received surgical treatment. The efficacy and cure rates were 100% (25/25) and 48% (12/25), respectively. The effective rate of aldactone and the single-drug cure rate were 89% (48/54) and 24% (13/54), respectively. Primary aldosteronism affects over 10% of hypertensive patients in China. The PAC/PRA ratio can be considered as a routine screening test in hypertensives, especially resistant hypertensive patients and a high PAC/PRA ratio is an invaluable index in primary aldosteronism diagnosis.
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The early percutaneous coronary intervention in elderly patients with acute coronary syndrome
HUANG Xiong, CAO Xuebin, ZHANG Gang
Front. Med.. 2008, 2 (1): 15-18.  
https://doi.org/10.1007/s11684-008-0004-3

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It is challenging to undo early percutaneous intervention (PCI) in the elderly with acute coronary syndrome (ACS). Fifteen patients older than 65 years with ACS within 24 hours of the event were admitted from April 4, 2004 to December 12, 2005. All the patients had early percutaneous coronary intervention and were followed up for 6–12 months by telephone or in the out-patient department. Nine of the 15 patients exhibited acute myocardial infarction (AMI). Six exhibited unstable angina (UA). All the patients had early PCI. The average door-to-balloon time was 78 minutes (40–110 minutes). The average PCI time was 99 minutes (68–120 minutes). Nineteen of 36 lesions in the fifteen angioplasty patients were treated and 20 stents were implanted in total. All the procedures were considered successful. Neither deaths nor recurrent angina occurred in the 6–12 months of follow-up. It was shown that early PCI might be an effective and safe method to treat elderly patients with ACS.
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Effects of RNA interference targeting angiotensin 1a receptor on blood pressure and cardiac hypertrophy of rats with renovascular hypertension
ZHANG Jingqun, SUN Honglei, MA Yexin, WANG Daowen
Front. Med.. 2008, 2 (1): 19-24.  
https://doi.org/10.1007/s11684-008-0005-2

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The aim of this study is to investigate the effects of RNA interference (RNAi) targeting angiotensin 1a receptor (AT1a) on blood pressure and cardiac hypertrophy of rats with renovascular hypertension. Two RNAi plasmids, pAT1a-shRNA1 and pAT1a-shRNA2 each carrying a U6 promoter and an AT1a-specific shRNA-coding template sequence corresponding to the sites 928–946, 978–996 of the mRNA transcript, and a control plasmid pCon carrying a nonspecific shRNA-coding sequence were constructed. Thirty Sprague – Dawley rats with renovascular hypertension (2-kidney 1-clip) were randomly divided into 5 equal groups: Control group (without any intervention), pAT1a-shRNA1, pAT1a-shRNA2, pCon groups (with injection of the corresponding plasmid 4 mg/kg respectively into the tail vein), and valsartan group (30 mg/kg·d-1 by gavage). Three weeks after drug administration, pAT1a-shRNA1, pAT1a-shRNA2 and valsartan respectively resulted in decrease of the tail blood pressure by (15.1 ± 5.4), (16.4 ± 8.4) and (30.6 ± 18.2) mmHg. However, the tail blood pressure increased further by about 25 mmHg in both of pCon and control groups. The carotid artery pressures of pAT1a-shRNA1, pAT1a-shRNA2 and valsartan groups were all significantly lower than those of the control and pCon groups. The ratio of left ventricle weight to body weight (LV/BW) of the rats in pAT1a-shRNA1, pAT1a-shRNA2, and valsartan groups decreased significantly than in the control group (P < 0.01), similar to those of the normal SD rats(P > 0.05). Histopathological examination showed that the myocardiocytes were significantly hypertrophic and the basal membrane of the aorta was significantly thickened in the control group and such changes were alleviated in the pAT1a-shRNA1, pAT1a-shRNA2 and valsartan groups. Compared with the control group, pAT1a-shRNA1 and pAT1a-shRNA2 groups had lowered expression of AT1 receptor (in the myocardium and the thoracic aorta (all P < 0.01); however, there were no significant differences in expression levels of AT1 receptor in valsartan and the control groups (P > 0.05). We conclude that RNAi targeting AT1a receptor inhibits the development of renovascular hypertension and the accompanying cardiac hypertrophy. RNAi technology may become a new strategy of gene therapy for hypertension.
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Expression of renal cubilin and its potential role in tubulointerstitial inflammation induced by albumin overload
YANG Jurong, HE Yani, SHEN Haiying, DING Hanlu, LI Kailong, WANG Huiming
Front. Med.. 2008, 2 (1): 25-34.  
https://doi.org/10.1007/s11684-008-0006-1

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Sustained proteinuria is an independent risk factor leading to kidney fibrosis and end-stage renal failure. Over-reabsorption of filtered proteins, notably albumin, has been proved to trigger interstitial inflammation and fibrosis in proteinuric renal disease. Cubilin, an endocytic receptor expressed on the renal tubular brush border, is responsible for albumin reabsorption in physiologic condition. However, little is known about whether it is required for activation of tubular cells induced by albumin overload. In this work, we investigated the change of cubilin expression and its potential role in albumin-induced up-regulation of chemokines synthesis in vivo and in vitro. Twenty-six patients with nephrotic syndrome were enrolled in this study. Proximal tubule uptake of albumin, expression of apical membrane cubilin and infiltrating cells in kidney interstitium were determined by immunocytochemistry. In vitro, the transcription of cubilin in HK2 cells after exposure to albumin was analyzed by real-time PCR. Endocytosis of albumin in HK2 cells was examined by fluorescent microscope. The influence of inhibition of cubilin on albumin-induced expressions of monocyte chemoattractant protein 1 (MCP-1) and regulated upon activation normal T-cell expressed and secreted (RANTES) was investigated by Western blot. The intensity of luminal cubilin and tubular accumulation of albumin were significantly increased in nephrotic kidneys. The expression of MCP-1 and RANTES was up-regulated, and there were spatial relationships in localization between these chemokines and cubilin as well as intracellular albumin in kidney tissues. Infiltration of CD-3 and ED-1-positive cells was predominant in tubulointerstitial areas displaying signs of increases of cubilin expression and albumin accumulation. In vitro, the transcription of cubilin mRNA in HK2 cells was enhanced after 24 h exposure to albumin in a dose-dependent manner. Inhibition of endocytosis of albumin by antisense cubilin nucleotide markedly reduced expression of MCP-1 and RANTES. Cubilin was required for handling a greater amount of protein in nephrotic status and albumin-induced production of MCP-1 and RANTES by renal tubular cells, which further initiated tubulointerstitial inflammation in proteinuric disease.
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Effects of podocin transfection on CD2AP distribution in HEK293 cells
SHA Yugen, HUANG Songming, ZHANG Aihua, ZHAO Fei, CHEN Ronghua
Front. Med.. 2008, 2 (1): 35-38.  
https://doi.org/10.1007/s11684-008-0007-0

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The aim of this paper is to construct a podocin fluorescence expression vector and observe the effects of podocin transfection on CD2AP distribution in HEK293 cells. The pGEMT-easy vector containing the full-length cDNA encoding human podocin was cloned and digested with BamHI and XhoI. The digested full-length podocin was subcloned into pEGFP-C2. The constructed plasmids were transfected into HEK293 cells and its effects on CD2AP distribution were observed by immunofluorescence. The pEGFP-NPHS2 expression vector was successfully constructed and podocin exclusively located on the HEK293 cell membrane. After podocin transfection, CD2AP redistributed from the perinucleus to the cytoplasm in HEK293 cells. It can be concluded that podocin can recruit CD2AP to redistribute from the perinucleus to the cytoplasm in HEK293 cells.
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Effects of Decoction on plasma proteome in cirrhosis: preliminary experimental study with rats
LIU Jie, WANG Jiyao, WEI Liming, LU Ye, Jin Hong
Front. Med.. 2008, 2 (1): 39-44.  
https://doi.org/10.1007/s11684-008-0008-z

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The aim of this paper is to study the effects of Fuzheng Huayu Decoction on the plasma proteome in cirrhotic rats. Twenty-six male Sprague-Dawley (SD) rats were randomly divided into three groups: cirrhotic model group (n = 10), treated with CCl4 (CCl4/olive oil: v/v = 1:1); Fuzheng Huayu Decoction intervention group (n = 10), treated with CCl4 + Fuzheng Huayu Decoction; and normal control group (n = 6), treated with olive oil only. After 8 weeks, blood samples were collected from the inferior vena cava to undergo bi-dimensional electrophoresis (2DE) and analysis by PDQuest 7.3 software. Differential protein spots were cut, enzyme hydrolysis was conducted, and peptide fragments extracted from the mixture underwent mass spectrometry (MS) with MALDI-TOF-TOF-MS. The liver fibrogenesis was assessed using a digital image analysis instrument of Masson’s trichrome stained sections. The fibrosis area of the Fuzheng Huayu Decoction was (8.9 ± 3.7)%, significantly smaller than that of the cirrhotic model group [(12.4 ± 4.7)%, P < 0.05]. Ten markedly changed protein spots were identified by MALDI-TOF-TOF-MS. Eight of the 10 proteins, including plasma glutathione peroxidase, plasma glutathione peroxidase precursor, prealbumin, haptoglobin, apolipoprotein A-IV precursor, complement C4, inter-alpha-inhibitor H4 heavy chain, and serine/threonine-protein kinase microtubule-affinity regulating kinase 1 (MARK1) were expressed very lowly in the cirrhotic model group while they were expressed highly in the Fuzheng Huayu Decoction group. The expression of liver regeneration-related protein LRRG03 and vimentin increased in the cirrhotic model group, and reduced in the Fuzheng Huayu Decoction group. Some proteins related to oxidative stress, cell proliferation and transformation have changed in the plasma of cirrhosis induced by CCl4. Fuzheng Huayu Decoction promotes protein synthesis and plays an anti-fibrotic role by anti-oxidation and accommodation of cell proliferation and transformation.
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Application of three-dimensional digitalized reconstruction of latissimus dorsi myocutaneous flap
PEI Guoxian, ZHANG Yuanzhi, LI Jianwei, JIN Dan, CHEN Jionghao, LI Yanbing, ZHONG Shizhen
Front. Med.. 2008, 2 (1): 45-50.  
https://doi.org/10.1007/s11684-008-0009-y

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Developments of digital technology and three-dimensional (3D) reconstruction allowed a precise description of anatomic structures. With the introduction of Visible Human Project and Virtual Chinese Human (VCH) techniques, more detailed anatomic images could be obtained. Digitized visible models of these structures can be applied as a useful tool in clinical training. The aim of this study was to reconstruct the normal structures of thoracodorsal artery in 3D images and to establish the digitized visible models of latissimus dorsi myocutaneous (LDM) flap. The cross-sectional images from the four VCH datasets were reviewed to study LDM and thoracodorsal artery structures on a section-by-section basis. Next, two adult fresh cadaver specimens were perfused with lead oxide-gelatine mixture and subject to radiographic CT scanning on their torsos. The cross-sectional images from the CT images were reviewed to study thoracodorsal artery structures. Three-dimensional computerized reconstructions of LDM flap structures were conducted from these datasets by using Amira 3.1 (TGS) software respectively. The 3D reconstructed visible models established from these datasets perfectly displayed the anatomic characteristics of LDM flap.
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Antitumor immunity of human SART3 gene vaccine against mouse tumor
HE Yu, YANG Shuhua, LIU Yong, LI Tao
Front. Med.. 2008, 2 (1): 51-57.  
https://doi.org/10.1007/s11684-008-0010-5

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To determine whether squamous cell carcinoma antigen recognized by human T cell 3 (SART3) gene can induce antitumor immunity against tumor cells which express the gene, we constructed mouse tumor cells expressing human SART3 (LM8-SART3) and carried out experiments in vitro/vivo. After subcutaneous injection with SART3 gene vaccine, cytotoxic T lymphocyte (CTL) activity in vitro was measured using Cell Counting Kit-8. As for the in vivo part, C3H mice were divided into several groups. One group was challenged with tumor cells after immunity. Another group was treated with the vaccine after tumor implantation. It was found that human SART3 DNA vaccine can elicit a specific CTL reaction from the mouse splenocytes. After vaccination, tumor occurrence and tumor growth speed was reduced. The vaccine also shows activity in tumor treatment. We conclude that the human SART3 DNA vaccine can induce antitumor ability against tumor cells expressing human SART3 (LM8-SART3) in vitro/vivo which may provide new possibilities in antitumor therapy.
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Diagnosis and treatment of thyroglossal duct carcinoma: Report of three cases with review of literatures
GAO Yan, CHEN Yan, SUN Zhipeng, GUO Chuanbin, YU Guangyan, ZHAN Yi
Front. Med.. 2008, 2 (1): 58-62.  
https://doi.org/10.1007/s11684-008-0011-4

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Thyroglossal duct carcinoma, which is usually diagnosed postoperatively, is a rare malignant tumor arising in the thyroglossal duct cyst. The definitive diagnosis can be made only after microscopic examination. We retrospectively reviewed three cases of thyroglossal duct carcinoma diagnosed in Peking University School and Hospital of Stomatology from January 1986 to August 2006. Clinical and pathological features were investigated and the optimal treatment protocol was proposed. The constituent ratio of thyroglossal duct carcinoma among surgically excised thyroglossal duct lesions was 2.9%. The clinical presentation of thyroglossal duct carcinoma was very similar to that of its benign counterpart. Two cases were diagnosed as thyroglossal duct cyst prior to the operation, the remaining one as dermoid cyst. All three cases were diagnosed as papillary carcinoma of thyroid origin after microscopic examination. Primary thyroglossal duct carcinoma should conform to the following criteria: localization of the carcinoma to a clearly demonstrable thyroglossal duct cyst or tract; clinically or histologically confirmed absence of carcinoma of the thyroid gland. Papillary carcinoma is the most common histological type, which usually develops slowly with an excellent prognosis. The histological characteristics including: formation of papillary structure; nuclear morphological variations such as ground glass nuclei, pseudoinclusions, intranuclear grooves and filaments; concentrically calcified structures termed psammoma bodies which is regarded as a strong indication of papillary carcinoma; and positivity in immunohistological staining for thyroglobin. Sistrunk procedure of excision is the choice for treatment. A close follow-up is needed. In the presence of thyroid gland masses or cervical lymphadenopathy, thyroidectomy or neck dissection should be recommended. The effect of thyroid suppression therapy and radioactive iodine therapy is not conclusive.
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Construction of an immortalized neural progenitor cell strain and analysis of its immunogenicity
AN Ke, XU Ying, TIAN Xuebi, GAO Feng, TIAN Yuke, YANG Hui, ZHANG Chuanhan
Front. Med.. 2008, 2 (1): 63-69.  
https://doi.org/10.1007/s11684-008-0012-3

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Neural progenitor cells (NPC) are those that are the source of neural cells for cell transplantation and gene therapy. The shortage in quantity and the limited life spans of primary cultured cells limit its widespread use in basic research. Immortalized NPC, which also possess the capacity of self-renewal and can proliferate infinitely, can produce a large number of NPCs with stable phenotype and genotype. Here we report that an immortalized neural progenitor cell strain, which we named as INPC, was successfully established by gene-transfer of simian virus 40 large T antigen gene mediated by liposomes. The INPC retained the biological characteristics of its original cells and provided a safe and reliable cell platform for the treatment of central nervous system diseases and transgenic cell transplantation. INPC could express low levels of MHC antigens which was down-regulated after differentiation. This indicates that INPC possesses poor immunogenicity. The immortalized cells may show good long-term survival and do not elicit an acute immunological response following transplantation.
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Protein aggregation in association with delayed neuronal death in rat model of brain ischemia
GE Pengfei, FU Shuanglin, LI Wenchen, WANG Chonghao, ZHOU Chuibing, LUO Yinan, LUO Tianfei
Front. Med.. 2008, 2 (1): 70-74.  
https://doi.org/10.1007/s11684-008-0013-2

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To investigate the relationship between protein aggregation and delayed neuronal death, we adopted rat models of 20 min ischemia. Brain ischemia was produced using the 2-vessel occlusion (2VO) model in rats Light microscopy, transmission electronic microscopy and Western blot analysis were performed for morphological analysis of neurons, and protein detection. The results showed delayed neuronal death took place at 72 h after ischemia-reperfusion, protein aggregates formed at 4 h after reperfusion and reached the peak at 24 h after reperfusion, and Western blot analysis was consistent with transmission electronic microscopy. We conclude that protein aggregation is one of the important factors leading to delayed neuronal death.
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Developmental expression of CAPON and Dexras1 in spinal cord of rats
LI Xin, CHENG Chun, CHEN Mengling, GAO Shangfeng, ZHAO Jian, NIU Shuqiong, SHEN Aiguo
Front. Med.. 2008, 2 (1): 75-81.  
https://doi.org/10.1007/s11684-008-0014-1

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To study the expression of the carboxy-terminal PSD-95/DLG/ZO-1 ligand of nNOS (CAPON) and Dexras1 mRNA during development in the spinal cord of rats, real-time polymerase chain reaction (Real-time PCR), as a quantitative method, was used to study the developmental expression of CAPON and Dexras1 mRNA level in the spinal cord. The spatial expression of CAPON and Dexras1 mRNA was examined by a combination of in situ hybridization (ISH) and immunofluorescence. During the development of the spinal cord, CAPON mRNA was expressed in low levels from embryo day 14 to day 18. At postnatal day 1, it reached the peak and was expressed in the part which will become the dorsal horn when mature. It then decreased gradually until postnatal week 12, when it presented in the ventral horn. At embryo day 14, Dexras1 mRNA was expressed at low levels, increased during embryo day 16 to day 18 and peaked at postnatal day 1. Spatiotemporal expression of Dexras1 mRNA was similar to CAPON as confirmed by correlation analysis and colocalization. CAPON and neuronal nitric oxide synthase (nNOS) was expressed within the same cells of the dorsal horn at postnatal day 1 but had different subcellular localizations. Co-expression of CAPON and Dexras1 mRNA in myeloid tissue during development process of rat indicates that the adaptor protein, CAPON may play a probable role in differentiation of neurons, synaptic plasticity and synaptogenesis by regulating nNOS to activate Dexras1.
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Prokaryotic expression and purification of fusion protein HSP70-EGFP and its application in the study of dendritic cells internalization antigen
QU Ping, SUI Yanfang, MA Jiahai, CHEN Guangsheng, LIU Libing, CHEN Jiankang, LIU Fang'e
Front. Med.. 2008, 2 (1): 82-86.  
https://doi.org/10.1007/s11684-008-0015-0

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To study the endocytic activity of dendritic cells (DCs) by obtaining fusion protein HSP70-EGFP as exogenous antigen and loading it with DCs derived from human peripheral blood. Fusion protein HSP70-EGFP was prokaryotically expressed, isolated and purified. DCs were isolated and cultured from human peripheral blood. The DCs were divided into 3 groups in the endocytic experiment. There were 106 DCs in each group. Group 1 and 2 were respectively incubated for 30 min. with HSP70-EGFP and EGFP. Group 3 was incubated with HSP70 for 30 min, and then incubated for 30 min. with HSP70-EGFP. Subsequently, 3 groups were placed in an incubator at 37°C for 0.5, 1, 2 and 24 h. Flow cytometry (FCM) was adopted to detect the amount of DCs with EGFP inside. IL-12 Eli-spot was adopted to detect the amount of DCs which secreted IL-12. There were 5 types in the experiment: LPS, inactive LPS, HSP70-EGFP, EGFP and no antigen. Fusion protein HSP70-EGFP was successfully obtained and its molecular weight was 97 000. It accounted for 35.32% of the total protein. Under irradiation of an ultraviolet lamp, the protein solution sent out viridescent fluorescence. The result detected by FCM indicated that after incubation for 0.5 h at 37°C, the positive rate in group 1 was 63%, while the other 2 groups were negative. After incubation for 1, 2 and 24 h at 37°C, the positive rates in the 3 groups were above 80%. The IL-12 Eli-spot examination shows that with HSP70-EGFP being loaded, the amount of DCs secreting IL-12 was 134.09 ± 31.78/105 cells, a little lower than that of DCs with LPS loaded (with the average point of 156.36 ± 15.73). There was no significant difference between the 2 groups (P < 0.01). By contrast, both of them were significantly higher than inactive LPS-(33.78 ± 1.40)/105 cells and EGFP-loaded (23.13 ± 4.57)/105 cells DC groups in the amount of DCs secreting IL-12 (P < 0.01). The results suggest that receptor-mediated phagocytosis plays a main role in the preliminary stage of DCs internalizing HSP70-EGFP. With increasing incubation time, pinocytosis begins to dominate. HSP70-EGFP may promote DCs to secret cell factor IL-12.
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Prognostic significance of clinicopathologic parameters in gastrointestinal stromal tumors: a study of 156 cases
LIANG Yumei, LI Xianghong, LU Youyong, LV Yali, ZHONG Mei, PU Xiaolu, LI Wenmei
Front. Med.. 2008, 2 (1): 87-94.  
https://doi.org/10.1007/s11684-008-0016-z

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The biological behavior of gastrointestinal stromal tumors (GISTs) are highly variable. To investigate the biological behavior of GIST, we collected 83 cases of gastric and 62 cases of small intestinal GIST from the Department of Pathology of the Chinese PLA General Hospital. The parameters include age, primary tumor location, tumor diameter, mitotic index, tumor necrosis, risk assessment, clinical stage and the c-kit exon 11 mutation. All these were analyzed in 105 cases along with the follow-up data and tested by log rank and COX hazard proportional model. We find that the average age of gastric GIST was 55.4 years. Of the 62 cases that were followed up, 17 cases had metastasis or recurrence and the 5-year survival rate was (66.51 ± 17.06)%. For the small intestinal GIST, the average age was 50.6 years and 43 cases were followed up. Of these, 22 cases had metastasis or recurrence and the 5-year survival rate was (61.76 ± 18.30)%. Small intestinal GIST was more frequently associated with metastasis and tumor relapse than gastric GIST (?2 = 6.131, P = 0.013). For gastric GIST, patients younger than 50 years (P = 0.046), the advanced clinical stage (P = 0.0001), the large tumor diameter (P = 0.0001), a high mitotic index (P = 0.0001), necrosis (P = 0.0001) and a high risk grade (P = 0.004) were all correlated with a lower survival rate. The COX hazard proportional model revealed that advanced clinical stage (P = 0.001), large tumor size (P = 0.001), a high mitotic index (P = 0.002) and the high risk grade (P = 0.018) indicated a poorer prognosis in gastric GIST. For small intestinal GIST, necrosis (P = 0.036) and advanced clinical stage (P = 0.010) were associated with lower survival rates and the clinical stage was shown to be an independent prognostic indicator. A total of 25 cases harbored mutations in c-kit exon 11. The frequency of c-kit mutation was 32% and 22.5% for gastric and small intestinal GIST, respectively. In gastric GIST, the mutated c-kit was predominant in patients older than 50 years of age. But in the small intestinal GIST, the mutated c-kit was predominant in the age group of 40–49 years. In conclusion, for gastric GIST, clinical stage, tumor size, mitotic index, and risk grade are the prognostic indicators. For small intestinal GIST, necrosis and clinical stage are the prognostic indicators. Small intestinal GIST are more aggressive than gastric GIST. The occurrence of c-kit mutation may correlate with the age of patients.
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Heterologous expression of signal protein 14-3-3 in and the subsequent immune response in mice
ZHENG Meijuan, SHEN Jilong, LUO Qingli, XU Yuanhong
Front. Med.. 2008, 2 (1): 95-99.  
https://doi.org/10.1007/s11684-008-0017-y

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Schistosomiasis japonica, a zoonosis caused by Schistosoma japonicum, is endemic to the Philippines and China. Several vaccine candidates have been identified and tested in different animal models, but it is still unclear which will be optimal for testing in the field. Therefore, new antigens and strategies are necessary for vaccine development against schistosomiasis japonica. The Sj14-3-3 gene was amplified and subcloned into the expression vector pPICZ?-B and transformed into P. pastoris X-33 by electroporation. Three transformants were induced with methanol. The cultural supernatant was collected and tested by SDS-PAGE and Western blotting. The protein of rSj14-3-3 was prepared and purified and BALB/c mice were immunized which was followed by a challenging infection. The immuno-protection was then evaluated. The Sj14-3-3 gene was expressed and secreted into the medium and its molecular weight was about 35000 as determined by SDS-PAGE. Western blotting showed that the protein had a high specificity against mouse-anti-Sj14-3-3 monoclonal antibody and rSj14-3-3 had a promising immune reactivity. The results of the immuno-protective experiments revealed that the worm reduction was 26.0%, 32.2%, and 36.8%, respectively. The number of eggs in liver tissue was reduced by 36.8%, 43.2%, and 46.1%, respectively. The recombinant Sj14-3-3 of eukaryotic expression in Pichia pastoris was successfully harvested. The molecular vaccine of Sj14-3-3 could partially induce resistance to the infection with S. japonicum in BALB/c mice. The recombinant protein Sj14-3-3 has promising immunological potentials for further approach to the diagnosis and development of molecular vaccine.
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Site-directed mutagenesis of long QT syndrome KCNQ1 gene
LI Wei, WANG Bin, XU Qiumei, KE Qinmei, YANG Junguo, DU Rong, TIAN Li, WANG Qing
Front. Med.. 2008, 2 (1): 100-104.  
https://doi.org/10.1007/s11684-008-0018-x

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To construct a polymerase chain reaction (PCR) site-directed mutagenesis of the long QT syndrome KCNQ1 gene in vitro, two sets of primers were designed according to the sequence of KCNQ1 cDNA and a mismatch was introduced into primers. Mutagenesis was performed in a two-step PCR. The amplified fragments from the third PCR which contained the mutation site were sub-cloned into the T-vector pCR2.1. Then, the fragments containing the mutation site was obtained from pCR2.1 using restriction enzymes digestion and inserted into the same restriction site of pIRES2-EGFP-KCNQ1. The sequencing analysis shows that the mutation site was correct. Mutation from A to G in site 983 of KCNQ1 cDNA was found. Using the Effectene transfection reagent, pIRES2-EGFP-KCNQ1 (G983A) was transfected into HEK cells successfully. These results may shed light on further functional study of KCNQ1 gene.
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Genetics analysis of haptoglobin gene in Fujian Han nationality
ZOU Qilian, LIN Ying, LIN Xiangquan, CHEN Yuanzhong
Front. Med.. 2008, 2 (1): 105-107.  
https://doi.org/10.1007/s11684-008-0019-9

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To study the genetic features (characteristics) of haptoglobin gene, four different age groups of Fujian Han people were investigated. The phenotypes of the haptoglobin of four different groups were analyzed by using polyacrylamide gel electrophoresis. The frequency of Hp1 in the population of Fujian Han nationality accounted for 0.340, among which children, youths, middle aged and elder groups were 0.307, 0.338, 0.363 and 0.383, respectively. The Hp0–0 phenotype frequency was 0.026 in which the four age groups accounted for 0.032, 0.046, 0.014 and 0.014, respectively. The frequency of Hp1 gene is rising with increasing age. The frequency of Hp0–0 phenotype is highest in the middle aged group and then tends to drop with increasing age.
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The correlated study of hyperuricemia and metabolic syndromes among males of Han ethnicity in the Xinjiang Uygur Autonomous Region, China
YAO Hua, SUN Yuping, LI Qing, YAO Wenhai, WANG Qiuyun, HU Jing, ABUDUREHEMAN Gulibaha, TUO Ling, JIANG Yan
Front. Med.. 2008, 2 (1): 108-112.  
https://doi.org/10.1007/s11684-008-0020-3

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This study investigates the relationship between hyperuricemia and metabolic syndrome (MS) among males of Han ethnicity in the Xinjiang Uygur Autonomous Region, China. The blood samples were collected from January to May, 2006 in Urumqi. It included 1496 subjects with ages between 20 to 70 years. All these subjects had physical examinations, blood pressure (BP) measurement and analysis for serum uric acid (SUA), fasting blood sugar (FBS), serum triglyceride (TG), serum total cholesterol (TC), serum high density lipoprotein (HDL-C) and serum low density lipoprotein (LDL-C) as well as biochemistry assay. The Data were analyzed using the Pearson Chi-Square Test, Independent-Samples T-Test and the Mantel-Haenszel Test for linear trend, respectively. The results show that the prevalence of MS in the present study was 18%. Blood pressure, fasting blood sugar, body mass index (BMI), waist/hip ratio(WHR), TG, TC and LDL-C were significantly higher in the hyperuricemic group than in normal group and these parameters were strongly related to serum uric acid levels. The components of metabolic syndrome such as obesity, hyperglycemia, hypertension and dyslipidemia had a close correlation with SUA. The Odds ratios of these metabolic diseases in the hyperuricemic group were 3.097 times, 2.633 times, 2.226 times and 3.058 times of the normal SUA group, respectively. They all have a positive correlation with hyperuricemia. Hyperuricemia is closely linked to the various components of the metabolic syndrome. More emphasis should be put on the evolving control and prevalence of hyperuricemia and metabolic syndrome among males of Han ethnicity in Xinjiang.
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Characterization of Bacillus amyloliquefacien contaminating 75% alcohol disinfectant
ZHANG Wanming, YUAN Yuesha, BIAN Cangli, TU Xianyu, ZHANG Wen, HUANG Huqiang, WANG Lan
Front. Med.. 2008, 2 (1): 113-116.  
https://doi.org/10.1007/s11684-008-0021-2

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The clinical characterization of Bacillus amyloliquefacien contaminating 75% alcohol disinfectants were studied. The bacteria were cultured and observed by using bacterial examination under the hospital infection monitor. According to the regulations, the resistance of bacterial to physical and chemical factors was tested. Drug sensitivity tests for 20 commonly used medicines were carried out using a K-B method. The bacterial plasmids were analyzed using the Birnboim method. The bacteria were found after being cultured in the clinically-used 75% alcohol disinfectant fluid. Their total number was more than 800 cfu/mL and they were identified as Bacillus amyloliquefacien. The bacteria were also found to be resistant to boiling for 5 min. It grew well in 95% alcohol disinfectant and was insensitive to 84 disinfectant fluids containing chlorine (1000 mg/L) and such disinfectants as ozone. They were able to be sterilized better through routine ultraviolet exposure for 30 min or gas pressure. The bacteria contained a 2.5 kb plasmid and were sensitive to 13 drugs and insensitive to 7 drugs of the 20 drugs tested. It was suggested that alcohol disinfectant fluid was easily contaminated by Bacillus amyloliquefacien, and the bacteria was resistant to disinfectant fluids such as alcohol and 84 disinfectants
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