Showing papers by "Renji Hospital published in 2002"

[Expression of fusion proteins in beta(2)GP I gene-transfected HEp-2 cells and its clinical application].

Abstract: OBJECTIVE To establish an indirect immunofluorescent test so as to improve the sensitivity and specificity of examination of antibodies to beta(2)-glycoprotein METHODS Full-length beta(2)GP cDNA was obtained from human hepatocellular cancer cell line HepG2 by RT-PCR and cloned into the mammalian expression vector pEGFP-C1 The recombinant plasmid pEGFP-beta(2)GP was transfected into HEp-2 cells RT-PCR, immunoblotting (IBT), confocal fluorescence microscopy, and indirect immunofluorescent test (IIF) were used to confirm the expression, localization, and antigenicity of fusion protein of green fluorescent protein (GFP) Serum specimens from 19 patients suspected as with secondary antiphospholipid syndrome (APS), 1 patient diagnosed as with primary APS, and 10 normal persons were detected with IIF-IgG-beta(2)GP1, ELISA-IgG-ACL, and ELISA-IgG-beta(2)GP I simultaneously RESULTS (1) The HEp-beta(2)GP I cells thus obtained retained their ability of expression of beta(2)GP-I-GFP for more than ten generations This beta(2)GP-I-GFP showed the antigenicity of beta(2)GP-I with a characteristic feature (2) Seven of the 20 serum specimens from APS patients showed characteristic immunofluorescent pattern No serum specimen from normal persons showed immunofluorescent staining The comparison of results of the three methods showed that the concordance between IIF-IgG-beta(2)GP I and ELISA-IgG-beta(2)GP I was the most perfect (Kappa = 0886) (3) HEp-beta(2)GP I retained the immunofluorescent property of HEp-2 cell CONCLUSION As a new kind of substrate of IIF, beta(2)GP I transfectant can be used to detect anti-beta(2)GP-I antibodies Transfeted HEp-2 cells keep the immunofluorescent property of HEp-2 cells in IFANA test and can be used as substrate for routine IFANA detection

Effects of histone acetylation and DNA methylation on p21( WAF1) regulation.

Abstract: Cell cycle progression is regulated by interactions between cyclins and cyclin-dependent kinases (CDKs). p21(WAF1) is one of the CIP/KIP family which inhibits CDKs activity. Increased expression of p21(WAF1) may play an important role in the growth arrest induced in transformed cells. Although the stability of the p21( WAF1) mRNA could be altered by different signals, cell differentiation and numerous influencing factors. However, recent studies suggest that two known mechanisms of epigenesis, i.e.gene inactivation by methylation in promoter region and changes to an inactive chromatin by histone deacetylation, seem to be the best candidate mechanisms for inactivation of p21( WAF1). To date, almost no coding region p21(WAF1) mutations have been found in tumor cells, despite extensive screening of hundreds of various tumors. Hypermethylation of the p21(WAF1) promoter region may represent an alternative mechanism by which the p21(WAF1/CIP1) gene can be inactivated. The reduction of cellular DNMT protein levels also induces a corresponding rapid increase in the cell cycle regulator p21(WAF1) protein demonstrating a regulatory link between DNMT and p21(WAF1) which is independent of methylation of DNA. Both histone hyperacetylation and hypoacetylation appear to be important in the carcinoma process, and induction of the p21(WAF1) gene by histone hyperacetylation may be a mechanism by which dietary fiber prevents carcinogenesis. Here, we review the influence of histone acetylation and DNA methylation on p21(WAF1) transcription, and affection of pathways or factors associated such as p 53, E2A, Sp1 as well as several histone deacetylation inhibitors.

Anticarcinogenic effect of FTY720 in human prostate carcinoma DU145 cells: modulation of mitogenic signaling, FAK, cell-cycle entry and apoptosis.

Abstract: Despite the high frequency of prostate cancer, therapeutic options for advanced disease are limited to chemotherapy, radiation or hormonal therapy and eventually fail in all patients. Therefore, alternative approaches need to be developed. We previously reported that FTY720, a metabolite from Isaria sinclarii, is a unique antitumor agent for an androgen-independent prostate cancer cell line and requires caspase-3 activation in apoptosis. In our study, we have evaluated the effect of FTY720 on a family of mitogen-activated protein kinases (MAPKs), focal adhesion kinase (FAK), mitochondrial transmembrane potential, caspase-9 and caspase-8 and analyzed the expression of some cell-cycle regulator proteins in DU145 cells in order to understand the various antitumor effects of FTY720. Apoptosis was quantified by phosphatidylserine exposure. Activation of MAPKs, cleavage of caspase-9 and caspase-8, status of cyclin-dependent kinases (CDKs) and Cip1/p21, a cyclin-dependent kinase inhibitor, were evaluated by Western blot analysis, in addition to FAK and phospho-FAK immunoprecipitation and cell-cycle analysis by FACScan. We found that in DU145 cells, 40 microM FTY720 caused activation of p38 MAPK and the upstream kinase MKK3/MKK6 but not SAPK/JNK. Mitochondrial transmembrane potential, FAK and ERK1/2 were reduced while caspase-9 and caspase-8 were cleaved. The p38-specific inhibitor had no effect on apoptosis induced by FTY720, whereas z-VAD.FMK, a broad-spectrum caspase inhibitor, did not inhibit the p38 MAPK activation. An amount of 20 microM FTY720 resulted in G(1) arrest and a decrease of CDK2 as well as CDK4, whereas it induced Cip1/p21. FTY720 may exert anticarcinogenic effects against prostate cancer cells possibly involving modulation of mitogenic signaling, cell-cycle regulators, induction of G(1) arrest and apoptotic death in DU145 cells.

Helicobacter pylori associated gastric diseases and lymphoid tissue hyperplasia in gastric antral mucosa

Abstract: Aim: To investigate the relation between Helicobacter pylori associated gastroduodenal diseases and lymphoid tissue hyperplasia in the antral mucosa and to pursue its evolution after eradication of H pylori Methods: Gastric antral biopsy specimens were obtained from 438 patients with H pylori positive gastroduodenal diseases (185 chronic gastritis, 69 gastric ulcer, and 184 duodenal ulcer) and 50 H pylori negative healthy controls Lymphoid follicles and aggregates were counted and other pathological features were scored according to the updated Sydney system for classification of chronic gastritis After a course of anti-H pylori treatment, biopsy specimens were obtained at four to six weeks, 12 months, and 24 months in the chronic gastritis patient group Results: The total prevalence of lymphoid follicles and aggregates in the biopsies was 799% (350 of 438; 95% confidence intervals (CI), 076 to 084) The prevalence and density of lymphoid follicles and aggregates were significantly different in the various gastroduodenal diseases The highest prevalence (899%; 95% CI, 083 to 097) and density (082) of lymphoid follicles and aggregates occurred in patients with gastric ulcers The lowest prevalence of lymphoid follicles and aggregates was found in patients with chronic gastritis (746%; 95% CI, 068 to 081), and the lowest density of lymphoid follicles and aggregates (056) was seen in patients with duodenal ulcers The prevalence and density of lymphoid follicles and aggregates correlated strongly with the activity and severity of gastric antral mucosal inflammation The eradication of H pylori resulted in a decrease in the prevalence and density of lymphoid follicles and aggregates Conclusion: The prevalence and density of lymphoid follicles and aggregates in gastric antral mucosal biopsies correlated closely with H pylori infection

N-desulfated non-anticoagulant heparin inhibits leukocyte adhesion and transmigration in vitro and attenuates acute peritonitis and ischemia and reperfusion injury in vivo

Abstract: Objective: Heparin, a highly sulfated proteoglycan, is known to have strong anticoagulant and anti-inflammatory activities. Here we sought to generate a heparin derivative, which had a significantly lower anticoagulant activity while retaining its strong anti-inflammatory activity.¶Materials and methods: Heparin was chemically modified and this discrete set of the heparin derivatives was tested for their anticoagulant activities, such as activated partial thromboplastin time (APTT), and anti-inflammatory activities, such as leukocyte adhesion and transmigration in vitro and acute peritonitis and ischemia and reperfusion injury in vivo.¶Results: We found that an N-desulfated heparin had 188-fold (compared to heparin) and 32-fold (compared to low molecular weight heparin; LMWH) reductions of APTT. The N-desulfated heparin inhibited adhesion of human promyeloid HL-60 cells to the stimulated human umbilical vein endothelial cells (HUVECs) under a physiological shear stress. It also prevented the transmigration of human neutrophils through the monolayers of the stimulated HUVECs. Further, intravenous administration of this compound attenuated the peritoneal infiltration of neutrophils in a mouse model of acute peritonitis, and reduced tissue edema and leukocyte deposition in a rabbit ear model of ischemia and reperfusion injury.¶Conclusion: It is to our best knowledge that the N-desulfated heparin has the lowest anticoagulant activity among LMWH and chemically modified heparin derivatives, while preserving a potent anti-inflammatory activity. These combined properties appear to suggest it as a safer medicine for treatment of inflammation.

Multicenter randomized controlled trial of heat-killed Lactobacillus acidophilus LB in patients with chronic diarrhea

Abstract: Chronic diarrhea is a common bowel disorder; disturbance of intestinal microorganisms may play a role in its pathogenesis. This study assessed the clinical efficacy of lyophilized, heat-killedLactobacillus acidophilus LB versus living lactobacilli in the treatment of chronic diarrhea. One hundred thirty-seven patients with chronic diarrhea were randomly allocated to receive either a 4-week course of 2 capsules of Lacteol® Fort twice a day (Lacteol group, 69 patients) or a 4-week course of 5 chewable tablets of Lacidophilin® three times a day (Lacidophilin group, 64 patients). The frequency of stools was recorded quantitatively, and semiquantitative parameters such as stool consistency, abdominal pain, distortion, and feeling of incomplete evacuation were evaluated. At the second and fourth week of treatment, mean bowel frequency was significantly lower in the Lacteol group than in the Lacidophilin group (1.88±1.24 vs 2.64±1.12, 1.39±0.92 vs 2.19±1.05; P<.05). At the end of the treatment, the clinical symptoms were markedly improved in the Lacteol group, indicating thatL. acidophilus LB is more effective than living lactobacilli in the treatment of chronic diarrhea.

Interventional study of high dose folic acid in gastric carcinogenesis in beagles

Abstract: Background: A decrease in folic acid and subsequent DNA hypomethylation may be involved in gastric carcinogenesis. Epidemiological and nutritional studies have indicated that folate status modulates the risk of developing cancers. Aims: To investigate whether folic acid plays an important role in the chemoprevention of gastric carcinogenesis induced by N -ethyl- N -nitrosoguanidine (ENNG) in beagles. Methods: Sixteen male beagles were randomly divided into two groups: folic acid treated group and control group. In both groups beagles were fed ENNG 75 mg per day for eight months and in the treated group 20 mg folic acid was given to beagles for 15 months. Gastroscopy and biopsies were performed before and every 2–3 months after administration of ENNG until the end of the experiment. Histopathological lesions were diagnosed with regard to the criteria for human gastric mucosal biopsies. Serum and gastric mucosal tissue folic acid concentrations were measured. Results: In the control group, all beagles developed gastric cancer (8/8) compared with only 3/8 in the folic acid treated group (p Conclusions: Our results indicate that high dose folic acid plays an important role in the chemoprevention of gastric carcinogenesis induced by a chemical carcinogen ENNG in beagles.

Validation of a two-pool model for the kinetics of 2-microglobulin

Abstract: Secondary amyloidosis due to beta-2-microglobulin (β2-m) is a serious long-term complication in patients on regular dialysis therapy. β2-m can be considered a middle-molecule marker used to facilitate the assessment of dialysis efficacy. For this purpose, a validated model that calculates characteristic efficacy parameters, such as Kt/V, TAC and generation rate, is needed. There is general agreement that β2-m-kinetics should be described by a two-pool model, but little has been published to validate such an approach. We measured the β2-m concentration profiles of eight stable patients during hemodialysis (HD) at the start of treatment, after 30 minutes, after 60 minutes, and every hour until the end. Thereafter they were measured at 10-minute intervals for an hour. The dialyser clearances were determined from the plasma concentrations in front of and behind the dialyser twice during each session - after 1 hour, and 4 hours from the start of treatment. The kinetic parameters of a two-pool model (e.g. the compartment volumes V 1 and V 2 , the mass transfer coefficient K 1 2 and the generation rate G) were determined from the optimal fit of the measured concentration profile. The table below summarises the results by giving the mean and standard deviation for each parameter: V (liters) V 1 /V 2 V %TBW K 1 2 m(ml/min) G (mg/kg/day) 10.0 ′ 1.6 4.60 ′ 1.8 28.4 ′ 3.1 56.3 ′ 25.2 2.50 ′ 0.66 Inter-individual differences in V 1 /V 2 and K 1 2 were high, ranging from 2.5 to 10.0 for V 1 /V 2 and from 26 to 140 for K 1 2 . Error analysis suggested that these wide ranges were due to the method and that in reality the probable range of V is 25-36% of TBW, of V 1 /V 2 3.5-5.3, and of K 1 2 30-80 ml/min. With standard values for these three parameters (V = 30% of TBW, V 1 /V 2 = 4.4 and K 1 2 = 55 ml/m), equal for all patients, and their respective ranges, Kt/V can be calculated with a standard deviation of 13%. Kt/V > 1.2 secures the maximum possible β2-m removal with three HD treatments a week. Conclusions: The parameters of a two-pool model of β2-m kinetics can be derived from concentration profiles obtained under routine dialysis conditions, but accuracy is not completely satisfactory. Similar to the dialysis dose for urea (Kt/V u r e a ) the dialysis dose for β2-m (Kt/V β 2 - m ) can be calculated from the pre- and post-dialysis concentrations of β2-m, body weight, ultrafiltration and dialysis time. Kt/V β 2 - m > 1.2 secures the maximum possible removal of β2-m in HD with three sessions per week.

Significance of cyclooxygenase-2 expression in human primary hepatocellular carcinoma.

Abstract: AIM: To clarify the significance of cyclooxygenase-2 (COX-2) expression in human primary hepatocellular carcinoma (HCC) and adjacent nontumorous tissues. METHODS: The COX-2 protein and mRNA were investigated in 27 HCC tissues with adjacent nontumorous tissues, and 5 histologically normal liver tissues, using immunohistochemistry and in situ hybridization. RESULTS: The well-differentiated HCC expressed COX-2 protein (5.68 ± 1.19) more strongly than moderated HCC (3.43 ± 1.98) and poor differentiated HCC (3.33 ± 1.50) (P 0.05). The expression of COX-2 mRNA was observed in the cytoplasm of the cells of HCC and of the hepatocytes in adjacent nontumorous tissues in which COX-2 protein was positive. CONCLUSION: The overexpression of COX-2 in well-differentiated HCC suggests that COX-2 may play a role in the early stages of hepatocarcinogensis.

[Prophylactic and therapeutic effect of oxymatrine on D-galactosamine-induced rat liver fibrosis].

Abstract: OBJECTIVE To investigate the prophylactic and therapeutic effect of oxymatrine on experimental liver fibrosis and to reveal its mechanism. METHODS By establishing D-galactosamine-induced rat liver fibrosis model, we observed the effect of oxymatrine on serum and tissue biochemical indexes, content of liver hydroxyline, expression of TGF?1 mRNA and changes of tissue pathology. RESULTS There was a decline of liver hydroxyline and serum AST and ALT in oxymatrine group compared to those of the D-GalN group. The hydroxyline content in oxymatrine pretreatment group was (0.50 0.11)mug/mg compared with (0.99 0.14)mug/mg in D-GalN group (t=8.366, P<0.01). The content in oxymatrine treatment group was (0.44 0.04)mug/mg compared with 0.70 0.06 in D-GalN group (t=9.839, P<0.01). The SOD activity was (149.81 15.28) NU/mg in oxymatrine pretreatment group and (95.22 16.33) NU/mg in the model group (t=7.309, P<0.01); (157.68 19.54) NU/mg in the treatment group compared with (119.88 14.94) NU/mg in the model group (t=4.348, P<0.01). MDA in the pretreatment group was (2.06 0.17) nmol/mg, lower than (4.57 0.37) nmol/mg in the model group (t=17.529, P<0.01). In the treatment group, it was (1.76 0.24)nmol/mg, lower than (3.10 0.17) nmol/mg in the model group (t=12.697, P<0.01). TGF?1 mRNA reduced in the pretreatment and treatment groups as compared with that in the model group (0.21 0.01 vs 0.50 0.01, t=48.665, P<0.01; 0.18 0.02 vs 0.38 0.01, t=22.464, P<0.01). Electron microscopy showed that oxymatrine group had milder hepatocyte degeneration and less fibrosis accumulation than did the model group. Microscopy revealed wide septa expansion from the portal area to the central venous, piecemeal and confluent necrosis and pseudo-nodular formation in part of the lobular in the model group. While in oxymatrine group these lesions were much improved. CONCLUSIONS Oxymatrine shows prophylactic and therapeutic effect in D-galactosamine induced rat liver fibrosis. This is partly by protecting hepatocyte and suppressing fibrosis accumulation through anti-lipoperoxidation.

Effect of albumin infusion on preventing the deterioration of renal function in patients with spontaneous bacterial peritonitis

Abstract: OBJECTIVE: To determine whether plasma volume expansion with albumin could prevent impairment of renal function and reduce mortality in cirrhotic patients with either acute spontaneous bacterial peritonitis (SBP) or patients complicated with SBP. METHODS: A total of 112 patients was randomly allocated to two groups: 56 patients were allocated to be treated with ceftriaxone and 56 patients were allocated to treatment with ceftriaxone plus intra-venous albumin in 3 weeks. Serum creatinine and blood urea nitrogen levels were monitored. RESULTS: Of the 56 patients (group 2) treated with ceftriaxone and albumin, five patients had renal impairment. Of the 56 patients (group 1) treated with ceftriaxone alone, 19 had renal impairment. The incidence of renal impairment was significantly lower in patients treated with ceftriaxone and albumin (5/56; 10%) than in patients treated with ceftriaxone alone (19/56; 34%; P = 0.002). In addition, in-hospital mortality in group 2 was 10% (5/56), but was 33% (17/56) in group 1. Thus, in-hospital mortality was significantly reduced from 33% (17/56) in patients treated with ceftriaxone to 10% (5/56) in patients treated with ceftriaxone and albumin (P = 0.01). CONCLUSIONS: The addition of intravenous albumin to an antibiotic regimen reduces the incidence of renal impairment and mortality in cirrhotic patients with SBP.

[Association between Bcl-2 gene polymorphism with systemic lupus erythematosus].

Abstract: OBJECTIVE: To study the association between bcl-2 gene polymorphism and systemic lupus erythematosus (SLE) in Chinese and detect the synergism between IL-10 and bcl-2 genotypes in determining susceptibility to SLE METHODS: The peripheral white blood cells from 232 nuclear families of SLE and 106 ethnically matched normal controls were collected and the DNA extracted The allelic distribution of one microsatellite located in Bcl-2 gene was determined by using fluorescent-labeled primer and genotyping method RESULTS: (1) The distribution frequency of Bcl-2 alleles and its major genotypes were significantly different between SLE group and normal controls (P = 0001 and 00029 respectively) (2) Transmission disequilibrium existed in Bcl-2 alleles Bcl-2-195 bp allele was preferentially transmitted to affected offspring (t: non-t = 122:82, P = 00051) by TDT approach Individuals carrying specific genotypes with both Bcl-2-195 bp allele and IL-10G138 bp allele were at higher risk of developing SLE than those carrying only Bcl-2-195 bp allele or IL-10G138 bp allele (OR = 300, 177, 107 respectively) CONCLUSION: Bcl-2 gene polymorphism is associated with SLE and may be directly involved in the process of SLE or in linkage disequilibrium with some susceptibility loci nearby Synergistic effect may exist between IL-10 and bcl-2 genotypes in determining susceptibility to SLE

Comparison of Pgp- and MRP-Mediated Multidrug Resistance in Leukemia Cell Lines

Abstract: Drug resistance is a major cause of the failure of anticancer chemotherapy. Multidrug resistance is often caused by over-expression of the P-glycoprotein (Pgp) or the multidrug resistance—related protein (MRP). In the present study, we compared daunorubicin (DNR) accumulation, subcellular distribution, and the effect of modulators on drug accumulation and subcel-lular distribution in the Pgp-expressing K562 cell line and the MRP-expressing HL60 cell line using reverse-transcriptase polymerase chain reaction, MTT (3-[4, 5-dimethylthiazol-z-yl]-2,5-diphenyltetrazolium bromide) drug cytotoxicity assay, fluo-rocytometry, and confocal laser scanning microscopy. The 2 resistant cell lines exhibit similar levels of resistance to DNR and decreased drug accumulation. Altered drug subcellular distribution in the resistant cell lines compared to that in the sensitive cell lines was shown and, moreover, differences in drug distributions between the 2 resistant cell lines were found. DNR fluorescence in the resistant HL60 cell line was distributed into punctate regions in the cytoplasm; the nucleus and other cytoplasm were almost negative. In contrast, the resistant K562 cells showed a bright fluorescent signal located in the peripheral cytoplasm and perinuclear region; the nucleus and other cytoplasmic regions showed no signal. Use of the modulator verapamil increased drug accumulation and restored the altered subcellular distribution of the drug in the 2 resistant cell lines. The Golgi apparatus inhibitor brefeldin A had similar action in the resistant HL60 line but had little effect in the resistant K562 line. Therefore, our study suggested that there were differences between the 2 resistant cell lines in the compartments sequestering DNR.

Autofluorescence imaging analysis of gastric cancer

Abstract: AIMS: To investigate the characteristics of gastric cancer in autofluorescence images. METHODS: A double-channel laser scanning confocal microscope with an argon ion laser (excitation wavelength 488 nm) and helium−neon laser (excitation wavelength 543 nm) were used to detect autofluorescence from 16 gastric cancer tissue specimens and corresponding normal gastric tissue. RESULTS: Autofluorescence from normal gastric tissue produced a green-colored image. The intensity of red color increased obviously in all gastric cancer tissues (100%) after illumination and the tissues produced a reddish-brown-colored image. CONCLUSIONS: A reddish-brown image is characteristic of autofluorescence in gastric cancer detected by an argon ion laser and helium−neon laser with a double-channel laser scanning confocal microscope. Autofluorescence imaging analysis is useful in the diagnosis of gastric cancer.

Mechanism of action of Tripterygium Wilfordii polyglycoside on experimental endometriosis

Abstract: Purpose of investigation: This study was designed to examine the therapeutic effectiveness and mechanism of action of Tripterygium Wilfordii polyglycoside (TWP) in the treatment of endometriosis. Methods: An experimental endometriosis model was developed using New Zealand White rabbits where endometrial tissue was autotransplanted into the peritoneum. Six weeks after transplantation, a total of 22 rabbits were randomly placed into two groups: Group 1 (n=17) was treated with TWP (10 mg/kg/day) and Group 2 (n=5) served as the water-fed control for three successive months. The volume of endometrial implants was measured before and after administration of TWP and water. Immune and endocrine systems were investigated in the normal phase, six weeks after induction of endometriosis, and three months after TWP treatment and water administration. Results: After treatment with TWP, the average volume of endometrial implants significantly decreased (p < 0.0001), and the antiendometrial antibody (EmAb) level decreased (p < 0.05) to near normal levels, but it did not decrease in the untreated controls. Serum FSH and LH levels also decreased after TWP treatment. Furthermore, electron microscopic examination of the pituitary ultra-structure revealed morphological changes in gonadotropic cells (G-cell) after treatment with TWP, and changes gradually disappeared four weeks after withdrawal of TWP. Conclusion: This study indicates that TWP has both hormonal and immune system action that is effective as a medical treatment for experimental endometriosis by modulating both reproductive endocrine functions and immunosuppression that results in remission of the disease.

[Relationship between type I autoimmune hepatitis and alleles of HLA-DRB1 in Chinese patients of Shanghai area].

Abstract: OBJECTIVE To analyze the association between alleles of HLA-DRB1 and type I autoimmune hepatitis (AIH) in patients from Shanghai, China METHODS In 32 Chinese patients with type I AIH and 48 healthy controls in Shanghai area, polymerase chain reaction amplification with sequence-specific primers (PCR-SSP) was performed to examine the association of the alleles of HLA-DRB1 and its subtypes with type I AIH RESULTS HLA-DRB1 typing by PCR-SSP showed that DR4 had a significantly increased frequency among patients with AIH versus healthy control (469% versus 208%; relative risk=335, chi(2)=599, P=0014) No other alleles differed significantly between the two groups In subtypes of DRB1*04, there was a trend for an increase in gene frequency of DRB1*0405 increased in patients with type I AIH versus healthy controls (219% vs 63%, chi(2)=423, P=004, but Pc=008) The frequency of arginine at position HLA-DR beta 71 of third hypervariable region significantly increased among patients with AIH versus healthy control (469% versus 188%, chi(2)=714, P=0008) CONCLUSIONS Type I AIH in Chinese patients of Shanghai area is associated with HLA-DR4 and arginine at position DR beta 71 of third hypervariable region

Affects of different access routes on autologous satellite cell implantation stimulating myocardial regeneration.

Abstract: Objective To study the effect of different access routes on autologous satellite cell implantation to stimulate myocardial regeneration. Methods Satellite cells were procured from skeletal muscle (gluteus max) of adult mongrel canine, cultured, proliferated and labeled with 4', 6-diamidino-2-phenylindone (DAPI) in vitro. The cells were autologously implanted into the site of acute myocardial infarction by local injection or perfusion through the ligated distal left anterior descending coronary artery. Specimens were harvested 2, 4 and 8 weeks later for histological study. Results The labeling efficiency of satellite cells with DAPI was close to 100%. Fluorescent cells were found at the infarcted zone, papillary muscle and local injection site. Some of these cells had progressively differentiated into striated muscle fibers connected to intercalated discs. The infant cells appeared different from the mature myocardium under an electron microscope. Satellite cells implanted by perfusion through the coronary artery were arranged in order of consistency with host myocardial fibers. The satellite cells, implanted by local injection, were found growing in a disordered way. Conclusion Satellite cells, implanted by coronary artery perfusion, can progressively differentiate into striated muscle fibers, arranging in order and disseminating over the infarcted zone. This approach seems more favorable for the recovery of myocardial contractile function than that of local injection.

Expression of lethal gene mRNA on placenta villi in patients with spontaneous abortion

Abstract: Objective To study the role of lethal gene: including retinoic X receptor(RXR)α, N-myc and trancript enhancer factor(TEF)-1 in human spontaneous abortion. Methods The levels of RXRα, N-myc and TEF-1 mRNA expression on placenta villi from 38 spontaneous abortion women and 33 normal early pregnant women were examined by reverse transcription, polymerase chain reaction. Results (1) The levels of RXRα mRNA on placenta villi from the abortion group were significantly decreased as compared with those from normal group (0.4±0.3 versus 0.6±0.3, P0.05); There were no significant differences in the levels of N-myc mRNA expression between the abortion group and normal group (2.1±1.2 versus 2.2±0.9, P0.05). The levels of TEF-1 mRNA on placenta from abortion group were significantly lower than those from normal group (1.6±1.1, 2.3±1.2, P0.05); (2) The levels of RXR α mRNA, TEF-1 mRNA in recurrent abortion group were significantly lower than those from non-recurrent group (0.3±0.2 versus 0.6±0.4, P0.05, 1.0±1.1 versus 1.9±1.2, P0.05). Conclusion It was suggested that the lower expression of RXR α mRNA, TEF-1 mRNA may play an important role in the pathogenesis of spontaneous abortion especially in recurrent abortion.

The effects of testosterone undecanoate on relaxation of rat corpus cavernosum smooth muscle in vitro

Abstract: OBJECTIVES To investigate the role of Undecanoate (Andriol), as a kind of testosterone, in regulating the relaxation of isolated rat corpus cavernosum in vitro METHODS The castrated rats were given high and low dosage Andriol respectively, compared with intact and castrated rats After treatment of 4 weeks, the corpora cavernosa were cut, trimmed as to strips Norepinephrine(NE) was added to contract each of the tissue strips Next, sodium nitroprusside(SNP), electric functional stimulation(EFS) were used to relax the strips Percent relaxations were examined RESULTS High-dose Andriol(20 mg/kg) was significantly effective to castrated rats on percent relaxation induced by 10(-3) mol/L SNP and EFS Low-dose Andriol(10 mg/kg) was also effective to relax strips induced by 10(-3) mol/L SNP According to statistics, the differences were significant (P < 001) CONCLUSIONS The percent relaxations of castrated rats were increased after taking Andriol, and it could increase the relaxation of corpara cavernosa

Vascular endothelial growth factor gene expression in patients' ischemic skeletal muscle with diabetic foot.

Abstract: OBJECTIVE To investigate the relationship between the endogenous vascular endothelial growth factor (VEGF) gene expression in diabetics' calf ischemic skeletal muscle and the pathogeny of diabetic foot. METHODS Twenty-four patients (33 limbs) were divided into 3 groups: diabetes mellitus (DM) without lower extremity ischemia (n = 5) (10 limbs); arteriosclerosis obliterans (ASO) without diabetes mellitus (n = 10) (13 limbs); diabetic lower limb arteriosclerosis obliterans (DLASO) (n = 9) (10 limbs). Control group consisted of normal volunteers (NOR) (n = 5) (10 limbs). The calf skeletal muscle tissue was obtained through muscle biopsy. RT-PCR was applied to determine the expression of hVEGF165mRNA. RESULTS There was no expression in the calf skeletal muscle tissue of normal volunteers and DM. The calf skeletal muscle tissue in DLASO had the expression of hVEGF165mRNA (0.021 +/- 0.013) micro g, but obviously lower than ASO (0.133 +/- 0.024) micro g, (t = 13.32, P < 0.01). CONCLUSIONS The endogenous VEGF gene expression in ischemic lower extremity of DLASO is obviously lower than that of ASO. It is the important endogenic cause of the genesis and development of diabetic foot ulcer.

Diagnosis of gastric cancer by using autofluorescence spectroscopy

Abstract: AIMS: To evaluate the clinical use of laser-induced fluorescence (LIF) spectra for diagnosing gastric cancer during gastroscopy. METHODS: A helium−cadmium laser system (excitation wavelength = 442 nm) was used to detect autofluorescence in 38 patients with endoscopically and histologically diagnosed gastric cancer and chronic gastritis. RESULTS: The LIF spectra produced by the helium− cadmium laser from gastric cancer tissue had one major peak (510 nm) and three minor peaks (590, 670, 710 nm). The intensity and shape of the LIF spectra in gastric cancer and normal gastric tissue were significantly different. Using the integral fluorescence intensities (FI) at 662−677 and 701−716 nm and the FI ratio of 510 nm/710 nm as the diagnostic parameters, it was possible to diagnose gastric cancer with a sensitivity of 83%, a specificity of 91%, a positive predictive value of 88% and a negative predictive value of 88%. The diagnostic accuracy was 88%. CONCLUSIONS: Using autofluorescence spectro­scopy to detect gastric cancer allows real-time detection of gastric cancerous lesions and it serves as a guide for taking biopsy specimens. The technique may become a useful tool in the diagnosis of early gastric cancer.

The A1166C polymorphism of the Angiotensin II type I receptor gene and the response to antihypertensive agents in the elderly

Abstract: Objective To investigate the distribution of A1166C polymorphism of the type Ⅰ angiotensin Ⅱ receptor (AT 1-R) gene in elderly hypertensive population and impact of polymorphism on the responese of BP after treatment with losartan and ACEI. Methods One hundred eighty six hypertensive subjects were randomly recruited from geriatric department in Shanghai. A→C polymorphic site at 1166 nucleotide position in the 3'-untranslated region of AT 1-R is determined using restriction length polymorphism analysis of DNA generated by polymerase chain reaction(PCR-RFLP). Sixty hypertensive subjects with average age of 71±6 yrs are divided into 2 groups according to genotypes and randomized to either an losartan or ACEI. Fifty-seven patients (AA=41,AC=16) were follow-up for 24 wks. Results (1)The frequency of C allele is 0.062. (2)Losartan decreased 24 hr,daytime,nighttime SBP by 7.4%,7.6%,and 6.8% and DBP by 6.5%,6.4%,7.4%,respectively in AA homoxygotes. ACEI decreased 24 h,daytime SBP by 6.2%,7.1% in AA homoxygotes. In AC hetrozygotes, after losartan treatment,the 24 hr SBP, daytime SBP, DBP were reduced by 5.2%, 5.6%, 5.6% in AC hetrozygotes. After ACEI treatment, the 24 hr SBP daytime SBP were reduced by 5.0%,5.8% in AC hetrozygotes. 2×2 fractorial analysis showed no interaction between genotypes and the effect of treatment. Conclusion A low allele frequency of A1166C was found in elderly Shanghai hypertensive. Although losartan and ACEI reduced SBP and DBP,no difference in response rate has been found in homozygotes and hetrozygotes. It seems the effect of losartan and ACEI on hypertension was independent from the genotypes of the patients with hypertension.

Effect of Helicobacter pylori infection on cyclooxygenase-2 expression in gastric antral mucosa

Abstract: OBJECTIVE: Helicobacter pylori infection is a major etiological cause of chronic gastritis. Inducible cyclooxygenase (COX-2) is an important regulator of mucosal inflammation. Recent studies indicate that expression of COX-2 may contribute to gastro­intestinal carcinogenesis. The aim of this study was to investigate the effects of H. pylori infection and eradication therapy on COX-2 expression in gastric antral mucosa. METHODS: Antral biopsies were taken from 46 H. pylori-infected patients, who also had chronic gastritis, both before and after anti-H. pylori treatment. The COX-2 protein was stained by using immunohistochemical methods and COX-2 expression was quantified as the percentage of epithelial cells expressing COX-2. Gastritis and H. pylori infection status were graded according to the Sydney system. RESULTS: Cyclooxygenase-2 expression was detected in the cytoplasm of gastric antral epithelial cells both before and after the eradication of H. pylori. Cyclooxygenase-2 expression in mucosa with H. pylori infection was compared with the corresponding mucosa after successful H. pylori eradication (20.1 ± 13.1%vs 13.8 ± 5.9%; P < 0.05). At the same time, COX-2 expression in H. pylori-infected mucosa was com­pared with the normal controls (18.0 ± 14.1%vs 12.3 ± 4.6%, P < 0.05). Expression of COX-2 was correlated with the degree of chronic inflammation (r= 0.78, P < 0.05). CONCLUSIONS: Our results showed that H. pylori infection leads to gastric mucosal overexpression of COX-2 protein, suggesting that the enzyme is involved in H. pylori-related gastric pathology in humans.

[Treatment for chronic dacryocystitis by dacryocystorhinotomy with Ho: YAG laser under endoscope].

Abstract: Objective To analyze the experience and curative effect in treating chronic dacrocystitis by dacryocystorhinotomy with Ho:YAG laser under endoscope. Method 20 patients of chronic dacryocystitis were treated by dacryocystorhinotomy with Ho:YAG laser under endoscope, followed-up for 1 approximately 2 years to observe the curative effects. Result One was unplugged 8 months after the operation, six after 6 months, four after 4 months and nine after 3 months. No recidivations was found one year after the operation and the effective rate was 100%. There was no complications in all of the cases. Conclusion The technique of dacryocystorhinotomy with Ho: YAG laser under endoscope overcomes the disadvantages of traditional method of operation.

Study of the effects of quercetin on PML gene and protein expression and localization in leukemia cells

Abstract: OBJECTIVE To investigate PML gene and protein expression and localization in leukemia cell lines. METHODS Cell morphology was assayed by Wright and fluorescence stain, PML mRNA expression by RT-PCR, and PML protein localization by immunofluorescence. RESULTS (1) Differentiation was observed by morphology in NB4 and HL-60 cells after treatment with all-trans retinoic acid (ATRA) while K562 cells did not show. Apoptosis was found in each cell line after treatment with quercetin. (2) After treatment with ATRA, the fusion protein disappeared and PML protein resumed in NB4 cells, while in HL-60 and K562 cells there was no difference from control cells. After treatment with quercetin, the fusion protein disappeared in NB4 cells, then degraded, and so did in HL-60 cells and K562 cells. (3) The expression of PML mRNA had no change in all the three cell lines after treatment with ATRA or quercetin. CONCLUSION PML plays a role of differentiation and apoptosis induction in leukemia cells at the translational level. PML in POD plays a role of apoptosis induction and growth control of leukemia cells.

Relevance of VacA and mucosal pathological changes in Chinese patients with upper gastrointestinal diseases before and after Helicobacter pylori eradication

Abstract: OBJECTIVE: To investigate the influence of VacA activity on gastric mucosa prior to and after Helicobacter pylori eradication in Chinese patients with peptic ulcers and chronic gastritis. METHODS: Seventy-four dyspeptic patients with H. pylori infection were enrolled. The status of H. pylori infection was evaluated by culture and histo­pathology before and 4−6 weeks after H. pylori eradication therapy. Histological specimens were examined and graded semiquantitatively according to the updated Sydney classification. RESULTS: Helicobacter pylori with VacA was found in 59 of 74 patients (80%), and its prevalence in patients with peptic ulcers and chronic gastritis was similar. Helicobacter pylori eradication rates in patients with VacA+ and VacA− strains were similar. Before eradication, the degrees of acute or chronic inflammation, epithelial damage, atrophy, intestinal metaplasia (IM) and the number of lymphoid follicles were similar in patients with VacA+ and VacA−H. pylori. Four to 6 weeks after the eradication of H. pylori infection, the degrees of acute and chronic inflammation, and epithelial damage in the antrum decreased significantly, particularly in patients with VacA+H. pylori (P < 0.0001). The number of lymphoid follicles in the antrum also decreased more in patients with VacA+H. pylori than in those with VacA−H. pylori (P= 0.051). However, there was no difference in the extent of atrophy and IM between these two groups. CONCLUSIONS: There is no specific correlation between VacA+/VacA−H. pylori strains and mucosal clinicopathological features in Chinese patients with upper gastrointestinal diseases before and after eradication therapy. Successful eradication of H. pylori infection does not improve atrophic and IM lesions of the gastric mucosa.

Effect of aging on male sexual function in 93 patients using international index of erectile function

Abstract: OBJECTIVES To investigate the influence of aging on male sexual function. METHODS The study selected 93 ED patients, aged from 23 to 64, who responded to the International Index of Erectile Function (IIEF) questionnaire. The questionnaire includes 15 items related to male sexual activity, which are organized into 5 domains, namely, erectile function (EF), orgasmic function (OF), sexual desire (SD), intercourse satisfaction (IS) and overall satisfaction (OS). For statistical analysis, ANOVA with DUNCAN test was conducted, and statistical significance was set at P < 0.05. Some other risk factors of ED such as hypertension, diabetes etc. had been excluded. RESULTS According to the age, the subjects were divided into 5 groups. With age increasing, the proportion of moderate and severe in each group increased from 16.17% to 57.14%, whereas EF score decreased from (19.50 +/- 4.64) to (15.27 +/- 5.64), OF score decreased from (6.93 +/- 2.86) to (5.62 +/- 2.94), SD score decreased from (6.33 +/- 1.63) to (4.50 +/- 2.94), IS score decreased from (10.17 +/- 1.94) to (6.93 +/- 2.90), OS score decreased from (5.00 +/- 0.89) to (3.15 +/- 1.84). The tendency took on linearity (P < 0.01). Aging was negatively correlated to above mentioned scores (r = 0.98, P < 0.01). CONCLUSION Aging could be thought as a risk factor of ED, which is negatively correlated with male's EF, OF, IS, OS and SD scores, furthermore. IIEF questionnaire is a useful tool assessing epidemiology of ED.