AIM: To investigate the location and expression of TIMP-1 and TIMP-2 in the liver of normal and experimental hepatic fibrosis in rats. METHODS: The rat models of experimental immunity hepatic fibrosis (n=20) were prep...AIM: To investigate the location and expression of TIMP-1 and TIMP-2 in the liver of normal and experimental hepatic fibrosis in rats. METHODS: The rat models of experimental immunity hepatic fibrosis (n=20) were prepared by the means of immunologic attacking with human serum albumin (HSA),and normal rats (n=10) served as control group. Both immunohistochemistry and in situ hybridization methods were respectively used to detect the TIMP-1 and TIMP-2 mRNA and related antigens in liver. The liver tissue was detected to find out the gene expression of TIMP-1 and TIMP-2 with RT-PCR. RESULTS: The TIMP-1 and TIMP-2 related antigens in livers of experimental group were expressed in myofibroblasts and fibroblasts (TIMP-1: 482±65 vs 60±20; TIMP-2:336±48 vs 50±19, P<0.001). This was the most obvious in portal area and fibrous septum. The positive signals were located in cytoplasm, not in nucleus. Such distribution and location were confirmed bysitu hybridization (TIMP-1/β-actin: 1.86±0.47 vs 0.36±0.08; TIMP-2/β-actin: 1.06±0.22 vs 0.36±0.08,P<0.001). The expression of TIMP-1 and TIMP-2 was seen in the liver of normal rats, but the expression level was very low. However, the expression of TIMP-1 and TIMP-2 in the liver of experimental group was obviously high. CONCLUSION: In the process of hepatic fibrosis, fibroblasts and myofibroblasts are the major cells that express TIMPs.The more serious the hepatic fibrosis is in the injured liver,the higher the level of TIMP-1 and TIMP-2 gene expression.展开更多
Stem cells are not only units of biological organization,responsible for the development and the regeneration oftissue and organ systems, but also are units in evolution bynatural selection. It is accepted that there ...Stem cells are not only units of biological organization,responsible for the development and the regeneration oftissue and organ systems, but also are units in evolution bynatural selection. It is accepted that there is stem cellpotential in the liver. Like most organs in a healthy adult,the liver maintains a perfect balance between cell gain andloss. It has three levels of cells that can respond to loss ofhepatocytes: (1) Mature hepatocytes, which proliferate afternormal liver tissue renewal, less severe liver damage, etc;they are numerous, unipotent, 'committed' and respondrapidly to liver injury. (2) Oval cells, which are activated toproliferate when the liver damage is extensive and chronic,or if proliferation of hepatocytes is inhibited; they lie withinor immediately adjacent tothe canal of Hering (CoH); theyare less numerous, bipotent and respond by longer, but stilllimited proliferation. (3) Exogenous liver stem cells, whichmay derive from circulating hematopoietic stem cells (HSCs)or bone marrow stem cells; they respond to allyl alcoholinjury or hepatocarcinogenesis; they are multipotent, rare,but have a very long proliferation potential. They make amore significant contribution to regeneration, and evencompletely restore normal function in a murine model ofhereditary tyrosinaemia. How these three stem cellpopulations integrate to achieve a homeostatic balanceremains enigmatic. This review focuses on the location,activation, markers of the three candidates of liver stemcell, and the most importantly, therapeutic potential ofhepatic stem cells.展开更多
AIM:To seek for an effective method to improve the immune responses induced by DNA vaccine expressing HBV surface antigen(pCR3.1-S)in Balb/c mice(H-2~d). METHODS:The pCR3.1-S plasmid and the eukaryotic expression vect...AIM:To seek for an effective method to improve the immune responses induced by DNA vaccine expressing HBV surface antigen(pCR3.1-S)in Balb/c mice(H-2~d). METHODS:The pCR3.1-S plasmid and the eukaryotic expression vectors expressing murine IL-2(pDOR-IL-2)or IL-12(pWRG3169)were injected into mice subcutaneously. The immune responses to pCR3.1-S and the adjuvant effect of the cytokines plasmid were studied.Meanwhile the effect of pCR3.1-S on anti-translated subcutaneous tumor of P815 mastocytoma cells stably expressing HBsAg(P815-HBV-S) was also studied.Anti-HBs in serum was detected by enzyme- linked immunoadsordent assay(ELISA)and HBsAg specific cytotoxic T lymphocytes(CTLs)activity was measured by ^(51)Cr release assay.After three weeks of DNA immunization,the cells of P815-HBV-S were inoculated into mice subcutaneously and the tumor growth was measured every five days.The survival rate and living periods of mice were also calculated. RESULTS:After 8 wk DNA immunization,the A 450 nm values of sera in mice immunized with pCR3.1,pCR3.1-S and pCR3.1-S codeliveried with IL-2 or IL-12 plasmids were 0.03+0.01,1.24±0.10,1.98±0.17 and 1.67±0.12 respectively.Data in mice codeliveried pCR3.1-S with IL-2 or IL-12 plasmids were significantly higher than that of mice injected pCR3.1 or pCR3.1-S only.The HBsAg specific CTL activities in mice coinjected with pCR3.1-S and IL-2 or IL- l2 eukaryotic expression vectors were(61.9±7.1)% and (73.3±8.8)%,which were significantly higher than that of mice injected with pCR3.1(10.1±2.1)% or pCR3.1-S(50.5 ±6.4)%.The HBsAg specific CTL activities in mice injected with pCR3.1,pCR3.1-S,pCR3.1-S combined with IL-2 or IL- l2 eukaryotic expression vectors decreased significantly to (3.2±0.8)%,(10.6±1.4)%,(13.6±1.3)% and(16.9±2.3) % respectively after the spleen cells were treated by anti- CD8^+ monoclonal antibody,but presented no significant change to anti-CD4^+ monoclonal antibody or unrelated to monoclonal antibody.The HBV-S DNA vaccine(pCR3.1-S) could evidently inhibit the tumor growth,prolong the survival period of mice and improve the survival rate of mice and these effects could be improved by IL-12 gene codeliveried. CONCLUSION:HBV DNA vaccine has a strong antigenicity in humoral and cellular immunities,which can be promoted by plasmid expressing IL-2 or IL-12.CD8+ cells executed the CTL activities.DNA vaccine may be useful for both prophylaxis and treatment of HBV infection.展开更多
It has been proved that severe acute respiratory syndrome (SARS) is caused by SARS-associated coronavirus, a novel coronavirus. SARS originated in Guangdong Province, the People's Republic of China at the end of 2...It has been proved that severe acute respiratory syndrome (SARS) is caused by SARS-associated coronavirus, a novel coronavirus. SARS originated in Guangdong Province, the People's Republic of China at the end of 2002. At present,it has spread to more than 33 countries or regions all over the world and affected 8 360 people and killed 764 by May 31,2003. Identification of the SARS causative agent and development of a diagnostic test are important. Detecting disease in its early stage, understanding its pathways of transmission and implementing specific prevention measures for the disease are dependent upon swift progress. Due to the efforts of the WHO-led network of laboratories testing for SARS, tests for the novel coronavirus have been developed with unprecedented speed. The genome sequence reveals that this coronavirus is only moderately related to other known coronaviruses. WHO established the definitions of suspected and confirmed and probable cases. But the laboratory tests and definitions are limited. Until now, the primary measures included isolation, ribavirin and corticosteroid therapy, mechanical ventilation, etc. Other therapies such as convalescent plasma are being explored. It is necessary to find more effective therapy. There still are many problems to be solved in the course of conquering SARS.展开更多
AIM: To investigate the regulatory effect of electroacupuncture (EA) at Zusanli (ST36) on tumor necrosis factor-alpha (TNFα) in rats with ulcerative colitis (UC), and further elucidate the therapeutic mechanism of EA...AIM: To investigate the regulatory effect of electroacupuncture (EA) at Zusanli (ST36) on tumor necrosis factor-alpha (TNFα) in rats with ulcerative colitis (UC), and further elucidate the therapeutic mechanism of EA on UC.METHODS: Thirty-two male Sprague-Dawley (SD) rats were randomly divided into four groups (n=-8): normal control group, UC control group, UC+ST36 group and UC+nonacupoint group. A solution containing ethanol and 2,4,6-trinitrobenzenesulfonic acid (TNBS) was instilled into the distal colon in the rat (at a dose of 100 mg/kg) to set up UC rat model. Rats in wakefulness state of UC+ST36 group were stimulated at ST36 by EA once a day, while those of UC+nonacupoint group were done at 0.5 cm beside ST36. After 10d treatment, all rats were sacrificed simultaneously. Colon musocal inflammation and damage were assessed by measuring colon mass, morphologic damage score, colonic myeloperoxidase enzyme (MPO) activity, serum TNF-α and colonic TNF-α mRNA level. Morphologic damage score was examined under stereomicroscope. Colonic MPO activity was measured by spectrophotometer method. Serum TNF-αconcentration was determined by radioimmunoassay (RIA).Colonic TNF-α mRNA expression level was analyzed by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).RESULTS: Ratio of colonic mass/body mass (mC/mB) and activity of colonic MPO (μkat/g tissue) markedly increased (8.5±2.6 vs 2.5±0.4; 145±25 vs 24±8, P＜0.01 vs normal control group). Compared with normal control rats, serum TNF-α and colonic TNF-α mRNA level in UC control group were increased 2.5 fold (2 278±170 vs 894±248, P＜0.01)and 4.3 fold (0.98±0.11 vs 0.23±0.11, P＜0.01)respectively. After EA at ST36, mc/mB and MPO activity were reduced significantly (5.3±2.0 vs 8.5±2.6; 104±36 vs145±25, P＜0.01, 0.05) compared with those of UC control group. Serum TNF-α and colonic TNF-α mRNA level were inhibited by EA stimulation at ST36 (P＜0.01). The inhibitory rate was 16 % and 44 % respectively.Morphologic damage score was also increased markedly in rat with UC (P＜0.01), whereas it was decreased by EA at ST36 (P＜0.05). There was no significant difference between UC control group and UC+EA at non-acupoint (P＞0.05). Furthermore, these parameters were highly correlated with each other (P＜0.01).CONCLUSION: Serum TNF-α concentration and colonic TNF-α mRNA expression level are increased significantly in UC rats in correlation with the severity of disease. It indicates that TNF-α is closely involved in the immune abnormalities and inflammatory responses in UC. EA at ST36 has therapeutic effect on UC by downregulating serum TNF-r and colonic TNF-r mRNA expression. High levels of TNF-αand its corresponding mRNA expression seem to be implicated in the pathogenesis of UC.展开更多
Severe acute respiratory syndrome (SARS), also called infectious atypical pneumonia, is an emerging infectious disease caused by a novel variant of coronavirus (SARS associated coronavirus, SARS-CoV). It is mainly cha...Severe acute respiratory syndrome (SARS), also called infectious atypical pneumonia, is an emerging infectious disease caused by a novel variant of coronavirus (SARS associated coronavirus, SARS-CoV). It is mainly characterized by pulmonary infection with a high infectivity and fatality.SARS is swept across almost all the continents of the globe, and has currently involved 33 countries and regions, including the China's Mainland, Hong Kong, Taiwan, North America and Europe. On June 30, 2003, an acumulative total reached 8450 cases with 810 deaths. SARS epidemic was very rampant in March, April and May 2003 in the mainland of China and Hong Kong. Chinese scientists and healthcare workers cooperated closely with other scientists from all over the world to fight the disease. On April 16, 2003, World Health Organization (WHO) formally declared that SARSCoV was an etiological agent of SARS. Currently, there is no specific and effective therapy and prevention method for SARS. The main treatments include corticosteroid therapy,antiviralagents, anti-infection, mechanical ventilation and isolation. This disease can be prevented and controlled, and it is also curable. Under the endeavor of the Chinese Government, medical staffs and other related professionals,SARS has been under control in China, and Chinese scientists have also made a great contribution to SARS research.Otherstudies in developing new detection assays and therapies, and discovering new drugs and vaccines are in progress. In this paper, we briefly review the current status of SARS in China.展开更多
AIM: To determine the ultrastructure of junction areas between neurons and astrocytes of supraoptic nuclei in rats orally administered 30 g/L NaCI solution for 5 days. METHODS: The anti-connexin (CX) 43 and anti-CX32 ...AIM: To determine the ultrastructure of junction areas between neurons and astrocytes of supraoptic nuclei in rats orally administered 30 g/L NaCI solution for 5 days. METHODS: The anti-connexin (CX) 43 and anti-CX32 double immunoelectromicroscopic labeled method, and anti-Fos or anti-glial fibdllary acidic protein (GFAP) immunohistochemistry were used to detect changes in the junctional area between neurons and astrocytes in supraoptic nuclei of 5 rats after 30 g/L NaCL solution was given for 5days. RESULTS: A heterotypic connexin32/connexin43 gapjunction (HGJ) between neurons and astrocytes (AS) in rat supraoptic nuclei was observed, which was characterized by the thickening and dark staining of cytomembranes with a narrow cleft between them. The number of HGJs and Fos like immunoreactive (-LI) cells was significantly increased following hyperosmotic stimuli, that is, the rats were administered 30 g/L NaCI solution orally or 90 g/L NaCI solution intravenously. HGJs could be blocked with carbenoxolone (CBX), a gap junction blocker, and the number of Fos-LI neurons was significantly decreased compared with that in rats without CBX injection, while Fos-LI ASs were not affected. CONCLUSION: HGJ may be a rapid adaptive signal structure between neurons and ASs in response to stimulation.展开更多
AIM: To analyse the role of genetic susceptibility and environmental factors in the process of esophageal cancer (EC) formation in Xi'an, China. METHODS: A hospital based case-control study, combined with molecula...AIM: To analyse the role of genetic susceptibility and environmental factors in the process of esophageal cancer (EC) formation in Xi'an, China. METHODS: A hospital based case-control study, combined with molecular epidemiological method, was carded out. A total of 127 EC cases and 101 controls were interviewed with questionnaires containing demographic items, habit of tobacco smoking, alcohol drinking, and family history of EC. Polymorphism of CYPIA1 and GSTM1 of 127 EC cases and 101 controls were detected by PCR method. The interactions between genetic susceptibility and environmental factors were also discussed. RESULTS: Tobacco smoking, alcohol drinking and a family history of EC were risk factors for EC with an OR of 2.04 (95% CI 1.15-3.60), 3.45(95% CI 1.74-6.91), 3.14 (95% CI 1.28-7.94), respectively. Individuals carrying CYP1A1 Val/Val genotype compared to those with CYP1A1 Ile/Ile genotype had an increased risk for EC (OR 3.35, 95% CI 1.49-7.61). GSTM1 deletion genotype was a risk factor for EC (OR1.81, 95% CI 1.03-3.18). Gene-environment interaction analysis showed that CYPIA1 Val/Val genotype, GSTM1 deletion genotype had synergetic interactions with tobacco smoking, alcohol drinking and family history of EC. CONCLUSION: Tobacco smoking, alcohol drinking and a family history of EC are risk factors for EC. CYP1A1 Val/Val and GSTM1 deletion genotypes are genetic susceptibility biomarkers for EC. There are synergic interactions between genetic susceptibility and environmental factors.展开更多
AIM: To establish a novel human hepatocellular carcinoma (HCC) cell line FHCC-98 from HCC tissue and to provide a suitable model for studying HCC occurrence, progress and metastasis.METHODS: Serially passaged cells we...AIM: To establish a novel human hepatocellular carcinoma (HCC) cell line FHCC-98 from HCC tissue and to provide a suitable model for studying HCC occurrence, progress and metastasis.METHODS: Serially passaged cells were cultured and their morphologies were observed under light and electron microscope. Cytogenetic study was conducted by using flow cytometry and chromosome analysis. Expressions of tumor markers such as α-fetoprotein (AFP), cytokeratin (CK) and hepatoma metastasis-associated factor HAb18G/CD147 on the FHCC-98 cells were detected by immunocytochemistry or Western blotting. Lactic dehydrogenase (LDH) isoenzymes were detected by polyacrylamide gel electrophoresis (PAGE).Xenograft was performed by inoculating FHCC-98 cells into the flanks of nude mice.RESULTS: Morphology of FHCC-98 cells was the same as that of other malignant cells. The expressions of the cells were positive for HAb18G/CD147 and CK, and negative for AFP. Its population doubling time was 21.4 h. The cell DNA was tetraploid and the major chromosomes were triploid by cytogenetics analysis. The tumorigenicity in nude mice was 100%. PAGE showed four bands representing LDH2, LDH3,LDH4 and LDH5.CONCLUSION: FHCC-98 is a novel HCC cell line and an ideal cell model for further exploring the mechanism of hepatocellular carcinoma invasion and metastasis.展开更多
AIM: To observe the effect of anti-tuberculosis therapy on liver function of pulmonary tuberculosis patients with hepatitis B virus (HBV) infection, and to compare the differences of liver function by two treatments o...AIM: To observe the effect of anti-tuberculosis therapy on liver function of pulmonary tuberculosis patients with hepatitis B virus (HBV) infection, and to compare the differences of liver function by two treatments of antituberculosis.METHODS: Forty-seven TB patients with HBV infection and 170 TB patients without HBV infection were divided into HPBE(S) and HLAMKO treatment groups. Liver function tests before and after the treatments were performed once in 2 wk or monthly, and their clinical manifestations were recorded.RESULTS: The rate of hepatotoxicity occurred in 26 (59%)TB patients with HBV during anti-TB treatment, higher than that in 40 (24%) TB patients without HBV. Hepatotoxicity occurred in 66 out of 217 patients, and the incidence of liver dysfunction was 46.1% in HPBE(S) group, significantly higher than that in HLAMKO group (12.7%) (P＜0.01).CONCLUSION: TB patients with HBV should choose HLAMKO treatment because of fewer hepatotoxicity.展开更多
AIM: To find a novel antigen (Ag) presentation strategy to improve the immune responses induced by dendritic cell (DC)vaccine expressing hepatitis C virus (HCV) core antigen (pcDNA3HCV C-Fc) in Balb/c mice (H-2d).METH...AIM: To find a novel antigen (Ag) presentation strategy to improve the immune responses induced by dendritic cell (DC)vaccine expressing hepatitis C virus (HCV) core antigen (pcDNA3HCV C-Fc) in Balb/c mice (H-2d).METHODS: pcDNA3HCV C-Fc plasmid and eukaryotic expression vector pcDNA3 were injected into mice sc. Immune responses to pcDNA3HCV C-Fc were studied. Meanwhile the effect of pcDNA3HCV C-Fc on anti-translated subcutaneous tumor of SP2/0 cells stably expressing HCV C Ag (SP2/0-HCV C-FC) was also studied. Anti-HCV C in serum was detected by enzyme-linked immunoadsordent assay (ELISA) and HCV specific cytotoxic T lymphocyte (CTL) activity was measured by LDH release assay. After 3 wk of DNA immunization,the cells of SP2/0-HCV C-FC were inoculated into mice subcutaneously and tumor growth was measured every 5 d.The survival rate and living time of mice were also calculated.RESULTS: After 4 wk of DC immunization, the A450 nm values of sera in mice immunized with pcDNA3HCV C-Fc-DC and pcDNA3-DC were 0.56±0.17 and 0.12±0.03 respectively. The antibody titres in mice codeliveried with pcDNA3HCV C-Fc with DC were significantly higher than those of mice injected with pcDNA3-DC. The HCV specific CTL activities in mice coinjected with DC and pcDNA3HCV C-Fc or empty expression vectors were(73.2±3.1) % and (24.4±8.8) %, which were significantly higher than those of mice injected with water.The DC vaccine could evidently inhibit tumor growth, prolong the survival time of mice and improve the survival rate of mice and these effects could be improved by HCV C-Fc (pcDNA3HCV C-Fc) gene codelivered.CONCLUSION: DC vaccine has a strong antigenicity in humoral and cellular immunities, which can be promoted by transduced pcDNA3HCV C-Fc expressing HCV C or Fc.Thus, pcDNA3HCV C-Fc-transduced DCs may be a promising candidate for a CTL-based vaccine against HCV.展开更多
AIM: To investigate the expression of gonadotropin-releasing hormone (GnRH) receptor and the effects of GnRH analog(alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats. METHODS: Immunohis...AIM: To investigate the expression of gonadotropin-releasing hormone (GnRH) receptor and the effects of GnRH analog(alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats. METHODS: Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of GnRH receptor in GSMC, respectively. Techniques of cell culture, OD value of MTT test, measure of ^3H-TdR incorporation, average fluorescent values of proliferating cell nuclear antigen (PCNA) and flow cytometric DNA analysis were used in the experiment. RESULTS: The cultured GSMC of rats showed immunoreactivity for GnRH receptor; positive staining was located in cytoplasm. GnRH receptor mRNA hybridized signals were also detected in cytoplasm. When alarelin (10^-9, 10^-7, 10^-5 mol/L) was administered into the medium and incubated for 24 h, OD value of MTT, ^3H-TdR incorporation and average fluorescent values of PCNA all decreased significantly as compared with the control group (P<0.05). The maximum inhibitory effect on cell proliferation was achieved a concentration of 10s mol/L and it acted in a dose-dependent manner. Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G1 phase and decrease ratio of S phase of GSMC of rats (P<0.05).The maximum inhibitory effect on ratio of S phase was at the concentration of 10-s mol/L and also acted in a dose-dependent manner. CONCLUSION: Our data suggest that GnRH receptor can be expressed by GSMC of rats. GnRH analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.展开更多
AIM:To evaluate the expression of transforming growthfactor-alpha (TGF-α) and hepatitis B surface antigen (HBsAg) in human hepatocellular carcinoma (HCC) tissues and its significance.METHODS:Seventy specimens of HCC ...AIM:To evaluate the expression of transforming growthfactor-alpha (TGF-α) and hepatitis B surface antigen (HBsAg) in human hepatocellular carcinoma (HCC) tissues and its significance.METHODS:Seventy specimens of HCC tissues were detected by immunohistochemical method. Five specimens of normal human liver tissues were used as control.RESULTS: The TGF-o~ positive expression rates in HCC and its surrounding tissues were 74.3%(52/70) and 88.1%(52/59), respectively. TGF-α positive granules were mainly in the cytoplasm and fewer existed on the karyotheca. The TGF-α positive expressing rate in well differentiated HCC was significantly higher than that in moderately and poorly differentiated HCC (P<0.05).The TGF-α positive expression also was observed in intrahepatic bile ducts (part of those were hyperplastic ducts).The HBsAg positive expression rates in HCC and its surrounding tissues were 21.4%(15/70) and 79.7%(47/59), respectively.HBsAg positive granules were in the cytoplasm, inclusion and on the karyotheca.There was a prominent positive correlation between TGF-α and HBsAg expression in HCC surrounding tissues (P<0.05,γ=0.34). TGF-α was usually existed with HBsAg in regenerated and/or dysplastic liver cells.In the five normal liver tissues, TGF-α and HBsAg were not detectable in hepatocytes and bile ducts.CONCLUSION:Hepatitis B virus infection is dosely related with hepatocarcinogenesis.The overexpression of TGF-α in the liver seems to be associated with the regeneration of hepatocytes injured by HBsAg.The continued expression of TGF-α might lead to dysplasia of liver cells and development of HCC. Furthermore, TGF-α might play a role in morphogenesis and regeneration of intrahepatic bile ducts.展开更多
AIM: To sum up the experience of the successful therapy for the severe hepatitis of pregnant woman with postpartum massive hemorrhage.METHODS: The advanced therapeutic methods including the bilateral uterine artery em...AIM: To sum up the experience of the successful therapy for the severe hepatitis of pregnant woman with postpartum massive hemorrhage.METHODS: The advanced therapeutic methods including the bilateral uterine artery embolism, hemodialysis and artificial liver support therapy were performed with comprehensive medical treatments and the course of the successful rescuing the patient was analyzed.RESULTS: Through the hospitalization of about two mouths the patient and her neonatus had gotten the best of care in our department and pediatric department separately. Both of them were discharged in good condition.CONCLUSION: The key points for a successful therapy of the pregnant woman with severe hepatitis are termination of the pregnancy and the control of their various complications. It was suggested that the proper combination of these measures of modern therapy would race against time for renewing of hepatic and renal functions.展开更多
AIM: To study the abnormal cytokeratin (CK) expression,emergence of CK19 with or without CK7, in liver parenchymal cells and the role of laminin (LN), a basement membrane protein, in this process.METHODS: Six hepatoce...AIM: To study the abnormal cytokeratin (CK) expression,emergence of CK19 with or without CK7, in liver parenchymal cells and the role of laminin (LN), a basement membrane protein, in this process.METHODS: Six hepatocellular carcinoma (HCC) cell lines were examined for different CKs, LN and its receptor by immunocytochemistry and Western blotting. Double immunofluorescent reaction, laser-scanning confocal microscopy and an in vitro induction procedure were used to demonstrate the role of LN in regulating CK19 expression in these cells.RESULTS: Immunoreactivities for CK8, CK18, CK7 and the receptor for LN were observed in all the six HCC cell lines examined. However, CK19 was merely found in four of the six cell lines, and was in any case associated with LN expression. Laser-scanning confocal microscopydemonstrated the concomitant presence of these two molecules in most of the positive cells. In the two HCC cell lines, originally negative for CK19, addition of LN to the culture medium resulted in an induction of CK19 in a dosedependent manner. Both the artificially induced and the intrinsic production of CK19 were completely blocked by an antibody to LN.CONCLUSION: LN can induce expression of CK19 in HCC cells in vitro, providing direct evidence for our hypothesis that the abnormal hepatocytic CK19 expression in situ is due to pathologic LN deposition.展开更多
AIM:To investigate the possible correlation of the expression of gonadotropin releasing hormone(GnRH) and its receptor with of proliferating cell nuclear antigen(PCNA) in human gastric adenocarcinoma cell proliferatio...AIM:To investigate the possible correlation of the expression of gonadotropin releasing hormone(GnRH) and its receptor with of proliferating cell nuclear antigen(PCNA) in human gastric adenocarcinoma cell proliferation and differentiation.METHODS:GnRH and its receptor and PCNA were detected in 30 gastric adenocarcinoma by immunohistochemical ABC technique. RESULTS:GnRH and its receptor had the same distribution pattern in gastric adenocarcinoma cells.The positive signal was found mainly in cytoplasm.The content of GnRH and its receptor immunoreative product in high differentiatied adenocarcinoma was significantly higher than those of the other two groups and the low differentiatied adenocarcinoma was the lowest(P< 0.05).PCNA positive signal which was found mainly in nucleus was gradually abated along with the raising of the extent of the differentiation.The differences were significant among the three groups(P< 0.05). In human gastric adenocarcinoma,the expression of GnRH and its receptor was negative correlation with the expression of PCNA(r=0.9).CONCLUSION:GnRH might be involved in the regulation of human gastric adenocarcinoma cell proliferation and differentiation.展开更多
AIM: To analyze a 30-year historical series of patients treated in our hospital, who ingested corrosive substances, and to assess the effectiveness of surgical therapy administered in patients with strictures after ca...AIM: To analyze a 30-year historical series of patients treated in our hospital, who ingested corrosive substances, and to assess the effectiveness of surgical therapy administered in patients with strictures after caustic injury in esophagus during this period.METHODS: A total of 79 cases of caustic burns in esophagus were treated in Tangdu Hospital from 1971 to 2001. Their clinical and pathological data were reviewed, and collected from the medical records of patients and interviews with them.RESULTS: More men (n = 61) than women (n = 18) ingested caustic substances with a sex ratio of 3.4:1 during the30-year period. The caustic materials were liquid lye and acids (54 cases and 25 cases, respectively). Sixty-eight patients were given esophageal replacement in more than three months after caustic injury with no postoperative death, of which 17 cases developed postoperative complications making a complication rate of 25%. The most common one was cervical anastomotic leakage. All patients had improvement in swallowing afterwards. CONCLUSION: The presence and severity of injuries are correlated with the amount of caustic substances ingested. Surgical treatment is a good option in patients with severe strictures, and colonic interposition might be the best surgical process. The most important factors to guarantee a successful outcome for surgery are good vascular supply and absence of tension in the anastomosis.展开更多
AIM: To develop a microarray-based prewarning system consisting of gastric cancer chip, prewarning data and analysis software for early detection of gastric cancer and pre-cancerous lesions. METHODS: Two high-density ...AIM: To develop a microarray-based prewarning system consisting of gastric cancer chip, prewarning data and analysis software for early detection of gastric cancer and pre-cancerous lesions. METHODS: Two high-density chips with 8 464 human cDNA sites were used to primarily identify potential genes specific for normal gastric mucosa, pre-cancerous lesion and gastric cancer. The low-density chips, composed of selected genes associated with normal gastric mucosa, precancerous lesion and gastric cancer, were fabricated and used to screen 150 specimens including 60 specimens of gastric cancer, 60 of pre-cancerous tissues and 30 of normal gastric mucosa. CAD software was used to screen out the relevant genes and their critical threshold values of expression levels distinguishing normal mucosa from pre-cancerous lesion and cancer. All data were stored in a computer database to establish a prewarning data library for gastric cancer. Two potential markers brcaal and ndr1 were identified by Western blot and immunohistochemistry. RESULTS: A total of 412 genes associated with three stages of gastric cancer development were identified. There were 216 genes displaying higher expression in gastric cancer, 85 genes displaying higher expression in pre-cancerous lesion and 88 genes displaying higher expression in normal gastric mucosa. Also 15 genes associated with metastasis of gastric cancer and 8 genes associated with risk factors were screened out for target genes of diagnosis chip of early gastric cancer. The threshold values of 412 selected genes to distinguish gastric cancer, pre-cancerous lesion from normal gastric mucosa were defined as 6.01±2.40, 4.86±1.94 and 5.42±2.17, respectively. These selected 412 genes and critical threshold values were compiled into an analysis software, which can automatically provide reports by analyzing the results of 412 genes obtained by examining gastric tissues. All data were compiled into a prewarning database for gastric cancer by CGO software. Northern blot and immunohistochemistry analysis confirmed that gene and protein of brcaa1 displayed lower expression in normal gastric mucosa and higher expression in gastric cancer tissues, conversely, ndr1 displayed lower expression in gastric cancer and higher expression in normal gastric mucosa. CONCLUSION: The microarray-based prewarning system for gastric cancer was developed. This system consisted of gastric cancer-associated gene chip, prewarning data and analysis software, which has a high potential for applications in the early detection of gastric cancer. The two potential markers brcaal and ndr1 identified may be used to distinguish cancer status fand non-cancer status.展开更多
AIM: To investigate how cholesterol (Ch) can affect thephenotype of bile duct fibroblasts of New Zealand rabbits.METHODS: 16 rabbits were divided randomly into twogroups: the control group and the experiment group. Th...AIM: To investigate how cholesterol (Ch) can affect thephenotype of bile duct fibroblasts of New Zealand rabbits.METHODS: 16 rabbits were divided randomly into twogroups: the control group and the experiment group. Therabbits in experiment group were fed with hypercholesteroldiet for 8 weeks. Bile duct was dissociated from rabbits andprepared for transmission electron microscopy. The purifiedbile duct fibroblasts were cultured and divided randomlyinto there groups: control group, Ch smiddle concentrationgroup (0.6 g/L), Ch high concentration group (1.2 g/L). Afterincubated for 72 h, the fibroblasts were made into specimensfor transmission electron microscopy. The expression of α-actin in bile duct fibroblasts was measured by means oflaser scanning confocal microscopy.RESULTS: With the transmission electron microscopy, thenormal bile duct fibroblasts were shuttle-shaped, and therewere abundant rough endoplasmic reticulums (RER), butfew mitochondria or microfilaments in cytoplasm. This isthe typical phenotype of fibroblasts. Bile duct fibroblasts ofhypercholesterolemic rabbits were observed, by thetransmission electron microscopy Rough endoplasmicreticulums were significantly reduced, with a lot ofmicrofilament bundles or stress fibers appeared in cytoplasm,especially under plasma membrane. Dense bodies werescattered within these bundles. Macula densas anddiscontinuous sarcolemma were found under plasmamembrane. It suggested that the bile duct fibroblasts ofhypercholesterolemic rabbits presented the phenotype ofsmooth muscle cell. The cultured bile duct fibroblasts alsohad typical phenotype of fibroblasts. After stimulated bymiddle concentration cholesterol (0.6 g/L) for 72 h, thereappeared lots of microfilaments in cytoplasm, but withoutdense body, macula densa and discontinuous sarcolemma.Observed with confocal microscopy, there were many regularbundles of microfilaments in fibroblasts treated with middleconcentration ch (0.6 g/L) and the expression of α-actinwas signifiantly increased. The average fluorescence valueof middle concentration group was 1 628+189 (P＜0.01 vscontrol group). Microfilaments and the expression of α-actinwere greatly decreased in fibroblastes of high concentrationgroup (1.2 g/L). The average fluorescence value of highconcentration group was 1 427±153 (P＜0.05 vs middleconcentration group). There were a lower expression of α-actin and few microfilaments in bile duct fibroblasts of controlgroup with an average fluorescence value of 1 224±138.CONCLUSION: Cholesterol can make bile duct fibroblastshave the phenotypic characteristics of smooth muscle cellboth in vitro andin vivo and this effect is more significant invivo. The effect is probably associated with some otherfactors besides cholesterol.展开更多
DC-SIGN, a dendritic Cell-specific adhesion receptor and a type Ⅱ transmembrane mannose-binding C-type lectin, is very important in the function of DC, both in mediating naive T cell interactions through ICAM-3 and a...DC-SIGN, a dendritic Cell-specific adhesion receptor and a type Ⅱ transmembrane mannose-binding C-type lectin, is very important in the function of DC, both in mediating naive T cell interactions through ICAM-3 and as a rolling receptor that mediates the DC-specific ICAM-2-dependent migration processes. It can be used by viral and bacterial pathogens including Human Immunodeficiency Virus (HIV), HCV, Ebola Virus, CMV and Mycobacterium tuberculosis to facilitate infection. Both DC-SIGN and DC-SIGNR can act either in cis, by concentrating virus on target cells, or in bans, by transmission of bound virus to a target cell expressing appropropriate entry receptors. Recent work showed that DC-SIGN are highaffinity binding receptors for HCV. Besides playing a role in entry into DC, HCV E2 interaction with DC-SIGN might also be detrimental for the interaction of DC with T cells during antigen presentation. The clinical strategies that target DCSIGN may be successful in restricting HCV dissemination and pathogenesis as well as directing the migration of DCs to manipulate appropriate immune responses in autoimmunity and tumorigenic situations.展开更多
基金Supported by the Postdoctoral Science Foundation of China,No.1999-10
文摘AIM: To investigate the location and expression of TIMP-1 and TIMP-2 in the liver of normal and experimental hepatic fibrosis in rats. METHODS: The rat models of experimental immunity hepatic fibrosis (n=20) were prepared by the means of immunologic attacking with human serum albumin (HSA),and normal rats (n=10) served as control group. Both immunohistochemistry and in situ hybridization methods were respectively used to detect the TIMP-1 and TIMP-2 mRNA and related antigens in liver. The liver tissue was detected to find out the gene expression of TIMP-1 and TIMP-2 with RT-PCR. RESULTS: The TIMP-1 and TIMP-2 related antigens in livers of experimental group were expressed in myofibroblasts and fibroblasts (TIMP-1: 482±65 vs 60±20; TIMP-2:336±48 vs 50±19, P<0.001). This was the most obvious in portal area and fibrous septum. The positive signals were located in cytoplasm, not in nucleus. Such distribution and location were confirmed bysitu hybridization (TIMP-1/β-actin: 1.86±0.47 vs 0.36±0.08; TIMP-2/β-actin: 1.06±0.22 vs 0.36±0.08,P<0.001). The expression of TIMP-1 and TIMP-2 was seen in the liver of normal rats, but the expression level was very low. However, the expression of TIMP-1 and TIMP-2 in the liver of experimental group was obviously high. CONCLUSION: In the process of hepatic fibrosis, fibroblasts and myofibroblasts are the major cells that express TIMPs.The more serious the hepatic fibrosis is in the injured liver,the higher the level of TIMP-1 and TIMP-2 gene expression.
文摘Stem cells are not only units of biological organization,responsible for the development and the regeneration oftissue and organ systems, but also are units in evolution bynatural selection. It is accepted that there is stem cellpotential in the liver. Like most organs in a healthy adult,the liver maintains a perfect balance between cell gain andloss. It has three levels of cells that can respond to loss ofhepatocytes: (1) Mature hepatocytes, which proliferate afternormal liver tissue renewal, less severe liver damage, etc;they are numerous, unipotent, 'committed' and respondrapidly to liver injury. (2) Oval cells, which are activated toproliferate when the liver damage is extensive and chronic,or if proliferation of hepatocytes is inhibited; they lie withinor immediately adjacent tothe canal of Hering (CoH); theyare less numerous, bipotent and respond by longer, but stilllimited proliferation. (3) Exogenous liver stem cells, whichmay derive from circulating hematopoietic stem cells (HSCs)or bone marrow stem cells; they respond to allyl alcoholinjury or hepatocarcinogenesis; they are multipotent, rare,but have a very long proliferation potential. They make amore significant contribution to regeneration, and evencompletely restore normal function in a murine model ofhereditary tyrosinaemia. How these three stem cellpopulations integrate to achieve a homeostatic balanceremains enigmatic. This review focuses on the location,activation, markers of the three candidates of liver stemcell, and the most importantly, therapeutic potential ofhepatic stem cells.
基金the National Natural Science Foundation of China, No.39770665
文摘AIM:To seek for an effective method to improve the immune responses induced by DNA vaccine expressing HBV surface antigen(pCR3.1-S)in Balb/c mice(H-2~d). METHODS:The pCR3.1-S plasmid and the eukaryotic expression vectors expressing murine IL-2(pDOR-IL-2)or IL-12(pWRG3169)were injected into mice subcutaneously. The immune responses to pCR3.1-S and the adjuvant effect of the cytokines plasmid were studied.Meanwhile the effect of pCR3.1-S on anti-translated subcutaneous tumor of P815 mastocytoma cells stably expressing HBsAg(P815-HBV-S) was also studied.Anti-HBs in serum was detected by enzyme- linked immunoadsordent assay(ELISA)and HBsAg specific cytotoxic T lymphocytes(CTLs)activity was measured by ^(51)Cr release assay.After three weeks of DNA immunization,the cells of P815-HBV-S were inoculated into mice subcutaneously and the tumor growth was measured every five days.The survival rate and living periods of mice were also calculated. RESULTS:After 8 wk DNA immunization,the A 450 nm values of sera in mice immunized with pCR3.1,pCR3.1-S and pCR3.1-S codeliveried with IL-2 or IL-12 plasmids were 0.03+0.01,1.24±0.10,1.98±0.17 and 1.67±0.12 respectively.Data in mice codeliveried pCR3.1-S with IL-2 or IL-12 plasmids were significantly higher than that of mice injected pCR3.1 or pCR3.1-S only.The HBsAg specific CTL activities in mice coinjected with pCR3.1-S and IL-2 or IL- l2 eukaryotic expression vectors were(61.9±7.1)% and (73.3±8.8)%,which were significantly higher than that of mice injected with pCR3.1(10.1±2.1)% or pCR3.1-S(50.5 ±6.4)%.The HBsAg specific CTL activities in mice injected with pCR3.1,pCR3.1-S,pCR3.1-S combined with IL-2 or IL- l2 eukaryotic expression vectors decreased significantly to (3.2±0.8)%,(10.6±1.4)%,(13.6±1.3)% and(16.9±2.3) % respectively after the spleen cells were treated by anti- CD8^+ monoclonal antibody,but presented no significant change to anti-CD4^+ monoclonal antibody or unrelated to monoclonal antibody.The HBV-S DNA vaccine(pCR3.1-S) could evidently inhibit the tumor growth,prolong the survival period of mice and improve the survival rate of mice and these effects could be improved by IL-12 gene codeliveried. CONCLUSION:HBV DNA vaccine has a strong antigenicity in humoral and cellular immunities,which can be promoted by plasmid expressing IL-2 or IL-12.CD8+ cells executed the CTL activities.DNA vaccine may be useful for both prophylaxis and treatment of HBV infection.
文摘It has been proved that severe acute respiratory syndrome (SARS) is caused by SARS-associated coronavirus, a novel coronavirus. SARS originated in Guangdong Province, the People's Republic of China at the end of 2002. At present,it has spread to more than 33 countries or regions all over the world and affected 8 360 people and killed 764 by May 31,2003. Identification of the SARS causative agent and development of a diagnostic test are important. Detecting disease in its early stage, understanding its pathways of transmission and implementing specific prevention measures for the disease are dependent upon swift progress. Due to the efforts of the WHO-led network of laboratories testing for SARS, tests for the novel coronavirus have been developed with unprecedented speed. The genome sequence reveals that this coronavirus is only moderately related to other known coronaviruses. WHO established the definitions of suspected and confirmed and probable cases. But the laboratory tests and definitions are limited. Until now, the primary measures included isolation, ribavirin and corticosteroid therapy, mechanical ventilation, etc. Other therapies such as convalescent plasma are being explored. It is necessary to find more effective therapy. There still are many problems to be solved in the course of conquering SARS.
基金the National Nature Science Foundation of China, No.39970888
文摘AIM: To investigate the regulatory effect of electroacupuncture (EA) at Zusanli (ST36) on tumor necrosis factor-alpha (TNFα) in rats with ulcerative colitis (UC), and further elucidate the therapeutic mechanism of EA on UC.METHODS: Thirty-two male Sprague-Dawley (SD) rats were randomly divided into four groups (n=-8): normal control group, UC control group, UC+ST36 group and UC+nonacupoint group. A solution containing ethanol and 2,4,6-trinitrobenzenesulfonic acid (TNBS) was instilled into the distal colon in the rat (at a dose of 100 mg/kg) to set up UC rat model. Rats in wakefulness state of UC+ST36 group were stimulated at ST36 by EA once a day, while those of UC+nonacupoint group were done at 0.5 cm beside ST36. After 10d treatment, all rats were sacrificed simultaneously. Colon musocal inflammation and damage were assessed by measuring colon mass, morphologic damage score, colonic myeloperoxidase enzyme (MPO) activity, serum TNF-α and colonic TNF-α mRNA level. Morphologic damage score was examined under stereomicroscope. Colonic MPO activity was measured by spectrophotometer method. Serum TNF-αconcentration was determined by radioimmunoassay (RIA).Colonic TNF-α mRNA expression level was analyzed by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).RESULTS: Ratio of colonic mass/body mass (mC/mB) and activity of colonic MPO (μkat/g tissue) markedly increased (8.5±2.6 vs 2.5±0.4; 145±25 vs 24±8, P＜0.01 vs normal control group). Compared with normal control rats, serum TNF-α and colonic TNF-α mRNA level in UC control group were increased 2.5 fold (2 278±170 vs 894±248, P＜0.01)and 4.3 fold (0.98±0.11 vs 0.23±0.11, P＜0.01)respectively. After EA at ST36, mc/mB and MPO activity were reduced significantly (5.3±2.0 vs 8.5±2.6; 104±36 vs145±25, P＜0.01, 0.05) compared with those of UC control group. Serum TNF-α and colonic TNF-α mRNA level were inhibited by EA stimulation at ST36 (P＜0.01). The inhibitory rate was 16 % and 44 % respectively.Morphologic damage score was also increased markedly in rat with UC (P＜0.01), whereas it was decreased by EA at ST36 (P＜0.05). There was no significant difference between UC control group and UC+EA at non-acupoint (P＞0.05). Furthermore, these parameters were highly correlated with each other (P＜0.01).CONCLUSION: Serum TNF-α concentration and colonic TNF-α mRNA expression level are increased significantly in UC rats in correlation with the severity of disease. It indicates that TNF-α is closely involved in the immune abnormalities and inflammatory responses in UC. EA at ST36 has therapeutic effect on UC by downregulating serum TNF-r and colonic TNF-r mRNA expression. High levels of TNF-αand its corresponding mRNA expression seem to be implicated in the pathogenesis of UC.
文摘Severe acute respiratory syndrome (SARS), also called infectious atypical pneumonia, is an emerging infectious disease caused by a novel variant of coronavirus (SARS associated coronavirus, SARS-CoV). It is mainly characterized by pulmonary infection with a high infectivity and fatality.SARS is swept across almost all the continents of the globe, and has currently involved 33 countries and regions, including the China's Mainland, Hong Kong, Taiwan, North America and Europe. On June 30, 2003, an acumulative total reached 8450 cases with 810 deaths. SARS epidemic was very rampant in March, April and May 2003 in the mainland of China and Hong Kong. Chinese scientists and healthcare workers cooperated closely with other scientists from all over the world to fight the disease. On April 16, 2003, World Health Organization (WHO) formally declared that SARSCoV was an etiological agent of SARS. Currently, there is no specific and effective therapy and prevention method for SARS. The main treatments include corticosteroid therapy,antiviralagents, anti-infection, mechanical ventilation and isolation. This disease can be prevented and controlled, and it is also curable. Under the endeavor of the Chinese Government, medical staffs and other related professionals,SARS has been under control in China, and Chinese scientists have also made a great contribution to SARS research.Otherstudies in developing new detection assays and therapies, and discovering new drugs and vaccines are in progress. In this paper, we briefly review the current status of SARS in China.
基金Supported by National Natural Science Foundation of China,No.39770251)The Fourth Military Medical University Foundation (CX01A024)
文摘AIM: To determine the ultrastructure of junction areas between neurons and astrocytes of supraoptic nuclei in rats orally administered 30 g/L NaCI solution for 5 days. METHODS: The anti-connexin (CX) 43 and anti-CX32 double immunoelectromicroscopic labeled method, and anti-Fos or anti-glial fibdllary acidic protein (GFAP) immunohistochemistry were used to detect changes in the junctional area between neurons and astrocytes in supraoptic nuclei of 5 rats after 30 g/L NaCL solution was given for 5days. RESULTS: A heterotypic connexin32/connexin43 gapjunction (HGJ) between neurons and astrocytes (AS) in rat supraoptic nuclei was observed, which was characterized by the thickening and dark staining of cytomembranes with a narrow cleft between them. The number of HGJs and Fos like immunoreactive (-LI) cells was significantly increased following hyperosmotic stimuli, that is, the rats were administered 30 g/L NaCI solution orally or 90 g/L NaCI solution intravenously. HGJs could be blocked with carbenoxolone (CBX), a gap junction blocker, and the number of Fos-LI neurons was significantly decreased compared with that in rats without CBX injection, while Fos-LI ASs were not affected. CONCLUSION: HGJ may be a rapid adaptive signal structure between neurons and ASs in response to stimulation.
基金Supported by the National Natural Science Foundation of China,No.39670651
文摘AIM: To analyse the role of genetic susceptibility and environmental factors in the process of esophageal cancer (EC) formation in Xi'an, China. METHODS: A hospital based case-control study, combined with molecular epidemiological method, was carded out. A total of 127 EC cases and 101 controls were interviewed with questionnaires containing demographic items, habit of tobacco smoking, alcohol drinking, and family history of EC. Polymorphism of CYPIA1 and GSTM1 of 127 EC cases and 101 controls were detected by PCR method. The interactions between genetic susceptibility and environmental factors were also discussed. RESULTS: Tobacco smoking, alcohol drinking and a family history of EC were risk factors for EC with an OR of 2.04 (95% CI 1.15-3.60), 3.45(95% CI 1.74-6.91), 3.14 (95% CI 1.28-7.94), respectively. Individuals carrying CYP1A1 Val/Val genotype compared to those with CYP1A1 Ile/Ile genotype had an increased risk for EC (OR 3.35, 95% CI 1.49-7.61). GSTM1 deletion genotype was a risk factor for EC (OR1.81, 95% CI 1.03-3.18). Gene-environment interaction analysis showed that CYPIA1 Val/Val genotype, GSTM1 deletion genotype had synergetic interactions with tobacco smoking, alcohol drinking and family history of EC. CONCLUSION: Tobacco smoking, alcohol drinking and a family history of EC are risk factors for EC. CYP1A1 Val/Val and GSTM1 deletion genotypes are genetic susceptibility biomarkers for EC. There are synergic interactions between genetic susceptibility and environmental factors.
基金Supported by the National High-Tech Research and Development Program of China,NO.2001AA215061
文摘AIM: To establish a novel human hepatocellular carcinoma (HCC) cell line FHCC-98 from HCC tissue and to provide a suitable model for studying HCC occurrence, progress and metastasis.METHODS: Serially passaged cells were cultured and their morphologies were observed under light and electron microscope. Cytogenetic study was conducted by using flow cytometry and chromosome analysis. Expressions of tumor markers such as α-fetoprotein (AFP), cytokeratin (CK) and hepatoma metastasis-associated factor HAb18G/CD147 on the FHCC-98 cells were detected by immunocytochemistry or Western blotting. Lactic dehydrogenase (LDH) isoenzymes were detected by polyacrylamide gel electrophoresis (PAGE).Xenograft was performed by inoculating FHCC-98 cells into the flanks of nude mice.RESULTS: Morphology of FHCC-98 cells was the same as that of other malignant cells. The expressions of the cells were positive for HAb18G/CD147 and CK, and negative for AFP. Its population doubling time was 21.4 h. The cell DNA was tetraploid and the major chromosomes were triploid by cytogenetics analysis. The tumorigenicity in nude mice was 100%. PAGE showed four bands representing LDH2, LDH3,LDH4 and LDH5.CONCLUSION: FHCC-98 is a novel HCC cell line and an ideal cell model for further exploring the mechanism of hepatocellular carcinoma invasion and metastasis.
文摘AIM: To observe the effect of anti-tuberculosis therapy on liver function of pulmonary tuberculosis patients with hepatitis B virus (HBV) infection, and to compare the differences of liver function by two treatments of antituberculosis.METHODS: Forty-seven TB patients with HBV infection and 170 TB patients without HBV infection were divided into HPBE(S) and HLAMKO treatment groups. Liver function tests before and after the treatments were performed once in 2 wk or monthly, and their clinical manifestations were recorded.RESULTS: The rate of hepatotoxicity occurred in 26 (59%)TB patients with HBV during anti-TB treatment, higher than that in 40 (24%) TB patients without HBV. Hepatotoxicity occurred in 66 out of 217 patients, and the incidence of liver dysfunction was 46.1% in HPBE(S) group, significantly higher than that in HLAMKO group (12.7%) (P＜0.01).CONCLUSION: TB patients with HBV should choose HLAMKO treatment because of fewer hepatotoxicity.
基金Supported by the National Natural Science Foundation of China, No.30170822
文摘AIM: To find a novel antigen (Ag) presentation strategy to improve the immune responses induced by dendritic cell (DC)vaccine expressing hepatitis C virus (HCV) core antigen (pcDNA3HCV C-Fc) in Balb/c mice (H-2d).METHODS: pcDNA3HCV C-Fc plasmid and eukaryotic expression vector pcDNA3 were injected into mice sc. Immune responses to pcDNA3HCV C-Fc were studied. Meanwhile the effect of pcDNA3HCV C-Fc on anti-translated subcutaneous tumor of SP2/0 cells stably expressing HCV C Ag (SP2/0-HCV C-FC) was also studied. Anti-HCV C in serum was detected by enzyme-linked immunoadsordent assay (ELISA) and HCV specific cytotoxic T lymphocyte (CTL) activity was measured by LDH release assay. After 3 wk of DNA immunization,the cells of SP2/0-HCV C-FC were inoculated into mice subcutaneously and tumor growth was measured every 5 d.The survival rate and living time of mice were also calculated.RESULTS: After 4 wk of DC immunization, the A450 nm values of sera in mice immunized with pcDNA3HCV C-Fc-DC and pcDNA3-DC were 0.56±0.17 and 0.12±0.03 respectively. The antibody titres in mice codeliveried with pcDNA3HCV C-Fc with DC were significantly higher than those of mice injected with pcDNA3-DC. The HCV specific CTL activities in mice coinjected with DC and pcDNA3HCV C-Fc or empty expression vectors were(73.2±3.1) % and (24.4±8.8) %, which were significantly higher than those of mice injected with water.The DC vaccine could evidently inhibit tumor growth, prolong the survival time of mice and improve the survival rate of mice and these effects could be improved by HCV C-Fc (pcDNA3HCV C-Fc) gene codelivered.CONCLUSION: DC vaccine has a strong antigenicity in humoral and cellular immunities, which can be promoted by transduced pcDNA3HCV C-Fc expressing HCV C or Fc.Thus, pcDNA3HCV C-Fc-transduced DCs may be a promising candidate for a CTL-based vaccine against HCV.
基金Supported by the Natural Science Foundation of China,No.39770388
文摘AIM: To investigate the expression of gonadotropin-releasing hormone (GnRH) receptor and the effects of GnRH analog(alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats. METHODS: Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of GnRH receptor in GSMC, respectively. Techniques of cell culture, OD value of MTT test, measure of ^3H-TdR incorporation, average fluorescent values of proliferating cell nuclear antigen (PCNA) and flow cytometric DNA analysis were used in the experiment. RESULTS: The cultured GSMC of rats showed immunoreactivity for GnRH receptor; positive staining was located in cytoplasm. GnRH receptor mRNA hybridized signals were also detected in cytoplasm. When alarelin (10^-9, 10^-7, 10^-5 mol/L) was administered into the medium and incubated for 24 h, OD value of MTT, ^3H-TdR incorporation and average fluorescent values of PCNA all decreased significantly as compared with the control group (P<0.05). The maximum inhibitory effect on cell proliferation was achieved a concentration of 10s mol/L and it acted in a dose-dependent manner. Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G1 phase and decrease ratio of S phase of GSMC of rats (P<0.05).The maximum inhibitory effect on ratio of S phase was at the concentration of 10-s mol/L and also acted in a dose-dependent manner. CONCLUSION: Our data suggest that GnRH receptor can be expressed by GSMC of rats. GnRH analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.
文摘AIM:To evaluate the expression of transforming growthfactor-alpha (TGF-α) and hepatitis B surface antigen (HBsAg) in human hepatocellular carcinoma (HCC) tissues and its significance.METHODS:Seventy specimens of HCC tissues were detected by immunohistochemical method. Five specimens of normal human liver tissues were used as control.RESULTS: The TGF-o~ positive expression rates in HCC and its surrounding tissues were 74.3%(52/70) and 88.1%(52/59), respectively. TGF-α positive granules were mainly in the cytoplasm and fewer existed on the karyotheca. The TGF-α positive expressing rate in well differentiated HCC was significantly higher than that in moderately and poorly differentiated HCC (P<0.05).The TGF-α positive expression also was observed in intrahepatic bile ducts (part of those were hyperplastic ducts).The HBsAg positive expression rates in HCC and its surrounding tissues were 21.4%(15/70) and 79.7%(47/59), respectively.HBsAg positive granules were in the cytoplasm, inclusion and on the karyotheca.There was a prominent positive correlation between TGF-α and HBsAg expression in HCC surrounding tissues (P<0.05,γ=0.34). TGF-α was usually existed with HBsAg in regenerated and/or dysplastic liver cells.In the five normal liver tissues, TGF-α and HBsAg were not detectable in hepatocytes and bile ducts.CONCLUSION:Hepatitis B virus infection is dosely related with hepatocarcinogenesis.The overexpression of TGF-α in the liver seems to be associated with the regeneration of hepatocytes injured by HBsAg.The continued expression of TGF-α might lead to dysplasia of liver cells and development of HCC. Furthermore, TGF-α might play a role in morphogenesis and regeneration of intrahepatic bile ducts.
文摘AIM: To sum up the experience of the successful therapy for the severe hepatitis of pregnant woman with postpartum massive hemorrhage.METHODS: The advanced therapeutic methods including the bilateral uterine artery embolism, hemodialysis and artificial liver support therapy were performed with comprehensive medical treatments and the course of the successful rescuing the patient was analyzed.RESULTS: Through the hospitalization of about two mouths the patient and her neonatus had gotten the best of care in our department and pediatric department separately. Both of them were discharged in good condition.CONCLUSION: The key points for a successful therapy of the pregnant woman with severe hepatitis are termination of the pregnancy and the control of their various complications. It was suggested that the proper combination of these measures of modern therapy would race against time for renewing of hepatic and renal functions.
基金the Natural Science Foundation of China(NSFC), Grants No.39470778 and No.30171052
文摘AIM: To study the abnormal cytokeratin (CK) expression,emergence of CK19 with or without CK7, in liver parenchymal cells and the role of laminin (LN), a basement membrane protein, in this process.METHODS: Six hepatocellular carcinoma (HCC) cell lines were examined for different CKs, LN and its receptor by immunocytochemistry and Western blotting. Double immunofluorescent reaction, laser-scanning confocal microscopy and an in vitro induction procedure were used to demonstrate the role of LN in regulating CK19 expression in these cells.RESULTS: Immunoreactivities for CK8, CK18, CK7 and the receptor for LN were observed in all the six HCC cell lines examined. However, CK19 was merely found in four of the six cell lines, and was in any case associated with LN expression. Laser-scanning confocal microscopydemonstrated the concomitant presence of these two molecules in most of the positive cells. In the two HCC cell lines, originally negative for CK19, addition of LN to the culture medium resulted in an induction of CK19 in a dosedependent manner. Both the artificially induced and the intrinsic production of CK19 were completely blocked by an antibody to LN.CONCLUSION: LN can induce expression of CK19 in HCC cells in vitro, providing direct evidence for our hypothesis that the abnormal hepatocytic CK19 expression in situ is due to pathologic LN deposition.
文摘AIM:To investigate the possible correlation of the expression of gonadotropin releasing hormone(GnRH) and its receptor with of proliferating cell nuclear antigen(PCNA) in human gastric adenocarcinoma cell proliferation and differentiation.METHODS:GnRH and its receptor and PCNA were detected in 30 gastric adenocarcinoma by immunohistochemical ABC technique. RESULTS:GnRH and its receptor had the same distribution pattern in gastric adenocarcinoma cells.The positive signal was found mainly in cytoplasm.The content of GnRH and its receptor immunoreative product in high differentiatied adenocarcinoma was significantly higher than those of the other two groups and the low differentiatied adenocarcinoma was the lowest(P< 0.05).PCNA positive signal which was found mainly in nucleus was gradually abated along with the raising of the extent of the differentiation.The differences were significant among the three groups(P< 0.05). In human gastric adenocarcinoma,the expression of GnRH and its receptor was negative correlation with the expression of PCNA(r=0.9).CONCLUSION:GnRH might be involved in the regulation of human gastric adenocarcinoma cell proliferation and differentiation.
文摘AIM: To analyze a 30-year historical series of patients treated in our hospital, who ingested corrosive substances, and to assess the effectiveness of surgical therapy administered in patients with strictures after caustic injury in esophagus during this period.METHODS: A total of 79 cases of caustic burns in esophagus were treated in Tangdu Hospital from 1971 to 2001. Their clinical and pathological data were reviewed, and collected from the medical records of patients and interviews with them.RESULTS: More men (n = 61) than women (n = 18) ingested caustic substances with a sex ratio of 3.4:1 during the30-year period. The caustic materials were liquid lye and acids (54 cases and 25 cases, respectively). Sixty-eight patients were given esophageal replacement in more than three months after caustic injury with no postoperative death, of which 17 cases developed postoperative complications making a complication rate of 25%. The most common one was cervical anastomotic leakage. All patients had improvement in swallowing afterwards. CONCLUSION: The presence and severity of injuries are correlated with the amount of caustic substances ingested. Surgical treatment is a good option in patients with severe strictures, and colonic interposition might be the best surgical process. The most important factors to guarantee a successful outcome for surgery are good vascular supply and absence of tension in the anastomosis.
基金Supported by The Max Planck Society, National Natural Science Foundation of China, No. 3990177 and 30070838Shaanxi Provincial Board of Public Health Focus Fund, No. 99ZH-002
文摘AIM: To develop a microarray-based prewarning system consisting of gastric cancer chip, prewarning data and analysis software for early detection of gastric cancer and pre-cancerous lesions. METHODS: Two high-density chips with 8 464 human cDNA sites were used to primarily identify potential genes specific for normal gastric mucosa, pre-cancerous lesion and gastric cancer. The low-density chips, composed of selected genes associated with normal gastric mucosa, precancerous lesion and gastric cancer, were fabricated and used to screen 150 specimens including 60 specimens of gastric cancer, 60 of pre-cancerous tissues and 30 of normal gastric mucosa. CAD software was used to screen out the relevant genes and their critical threshold values of expression levels distinguishing normal mucosa from pre-cancerous lesion and cancer. All data were stored in a computer database to establish a prewarning data library for gastric cancer. Two potential markers brcaal and ndr1 were identified by Western blot and immunohistochemistry. RESULTS: A total of 412 genes associated with three stages of gastric cancer development were identified. There were 216 genes displaying higher expression in gastric cancer, 85 genes displaying higher expression in pre-cancerous lesion and 88 genes displaying higher expression in normal gastric mucosa. Also 15 genes associated with metastasis of gastric cancer and 8 genes associated with risk factors were screened out for target genes of diagnosis chip of early gastric cancer. The threshold values of 412 selected genes to distinguish gastric cancer, pre-cancerous lesion from normal gastric mucosa were defined as 6.01±2.40, 4.86±1.94 and 5.42±2.17, respectively. These selected 412 genes and critical threshold values were compiled into an analysis software, which can automatically provide reports by analyzing the results of 412 genes obtained by examining gastric tissues. All data were compiled into a prewarning database for gastric cancer by CGO software. Northern blot and immunohistochemistry analysis confirmed that gene and protein of brcaa1 displayed lower expression in normal gastric mucosa and higher expression in gastric cancer tissues, conversely, ndr1 displayed lower expression in gastric cancer and higher expression in normal gastric mucosa. CONCLUSION: The microarray-based prewarning system for gastric cancer was developed. This system consisted of gastric cancer-associated gene chip, prewarning data and analysis software, which has a high potential for applications in the early detection of gastric cancer. The two potential markers brcaal and ndr1 identified may be used to distinguish cancer status fand non-cancer status.
文摘AIM: To investigate how cholesterol (Ch) can affect thephenotype of bile duct fibroblasts of New Zealand rabbits.METHODS: 16 rabbits were divided randomly into twogroups: the control group and the experiment group. Therabbits in experiment group were fed with hypercholesteroldiet for 8 weeks. Bile duct was dissociated from rabbits andprepared for transmission electron microscopy. The purifiedbile duct fibroblasts were cultured and divided randomlyinto there groups: control group, Ch smiddle concentrationgroup (0.6 g/L), Ch high concentration group (1.2 g/L). Afterincubated for 72 h, the fibroblasts were made into specimensfor transmission electron microscopy. The expression of α-actin in bile duct fibroblasts was measured by means oflaser scanning confocal microscopy.RESULTS: With the transmission electron microscopy, thenormal bile duct fibroblasts were shuttle-shaped, and therewere abundant rough endoplasmic reticulums (RER), butfew mitochondria or microfilaments in cytoplasm. This isthe typical phenotype of fibroblasts. Bile duct fibroblasts ofhypercholesterolemic rabbits were observed, by thetransmission electron microscopy Rough endoplasmicreticulums were significantly reduced, with a lot ofmicrofilament bundles or stress fibers appeared in cytoplasm,especially under plasma membrane. Dense bodies werescattered within these bundles. Macula densas anddiscontinuous sarcolemma were found under plasmamembrane. It suggested that the bile duct fibroblasts ofhypercholesterolemic rabbits presented the phenotype ofsmooth muscle cell. The cultured bile duct fibroblasts alsohad typical phenotype of fibroblasts. After stimulated bymiddle concentration cholesterol (0.6 g/L) for 72 h, thereappeared lots of microfilaments in cytoplasm, but withoutdense body, macula densa and discontinuous sarcolemma.Observed with confocal microscopy, there were many regularbundles of microfilaments in fibroblasts treated with middleconcentration ch (0.6 g/L) and the expression of α-actinwas signifiantly increased. The average fluorescence valueof middle concentration group was 1 628+189 (P＜0.01 vscontrol group). Microfilaments and the expression of α-actinwere greatly decreased in fibroblastes of high concentrationgroup (1.2 g/L). The average fluorescence value of highconcentration group was 1 427±153 (P＜0.05 vs middleconcentration group). There were a lower expression of α-actin and few microfilaments in bile duct fibroblasts of controlgroup with an average fluorescence value of 1 224±138.CONCLUSION: Cholesterol can make bile duct fibroblastshave the phenotypic characteristics of smooth muscle cellboth in vitro andin vivo and this effect is more significant invivo. The effect is probably associated with some otherfactors besides cholesterol.
基金Supported by the National Natural Science Foundation of China,No.30170822
文摘DC-SIGN, a dendritic Cell-specific adhesion receptor and a type Ⅱ transmembrane mannose-binding C-type lectin, is very important in the function of DC, both in mediating naive T cell interactions through ICAM-3 and as a rolling receptor that mediates the DC-specific ICAM-2-dependent migration processes. It can be used by viral and bacterial pathogens including Human Immunodeficiency Virus (HIV), HCV, Ebola Virus, CMV and Mycobacterium tuberculosis to facilitate infection. Both DC-SIGN and DC-SIGNR can act either in cis, by concentrating virus on target cells, or in bans, by transmission of bound virus to a target cell expressing appropropriate entry receptors. Recent work showed that DC-SIGN are highaffinity binding receptors for HCV. Besides playing a role in entry into DC, HCV E2 interaction with DC-SIGN might also be detrimental for the interaction of DC with T cells during antigen presentation. The clinical strategies that target DCSIGN may be successful in restricting HCV dissemination and pathogenesis as well as directing the migration of DCs to manipulate appropriate immune responses in autoimmunity and tumorigenic situations.
