Tumor angiogenesis and dynamic CT in colorectal carcinoma: Radiologic-pathologic correlation

AIM: To investigate the correlation between microvessel density and spiral CT perfusion imaging in colorectal carcinoma.METHODS: Thirty-seven patients, with histologically proven colorectal carcinoma, underwent water enema spiral CT scan. The largest axial surface of the primary tumor was searched on unenhanced spiral CT images. At this level, the enhanced dynamic scan series was acquired.Time-density curves (TDC) were created from the region of interest drawn over the tumor, target artery by Toshiba Xpress/SX spiral CT with perfusion functional software.Then the perfusion was calculated. Microvessel density(MVD) was evaluated using immunohistochemical staining of surgical specimens with anti-CD34, and then MVD was correlated with perfusion.RESULTS: MVD of colorectal carcinomas was 33.11-173.44,mean 87.28, and perfusion was 15.60-64.80 mL/min/100 g, mean 39.74 mL/min/100 g. MVD and perfusionwere not associated with invasive depth, metastasis and disease stage, and they all decreased with increasing Dukes' stage, but no significant correlation was found between them (r= 0.L8, P = 0.29).CONCLUSION: There is no significant correlation between MVD and perfusion. Neovascularizaton and perfusion are highly presented in early colorectal carcinoma.CT perfusion imaging may be more suited for assessing tumorigenesis in colorectal carcinoma than histological MVD technique.

Effect of androgen deprivation on penile ultrastructure

<abstract>Aim: To investigate the ultrastructural changes of penile corpus cavernosum and tunica albuginea in rats treated with castration or finasteride. Methods: Eighteen male Sprague-Dawley rats of nine weeks old were randomly divided into three groups with 6 rats each. Group A served as the control, Group B was castrated and Group C, treated with finasteride. Four weeks later, rats were anesthetized and blood samples obtained for the determination of serum testosterone (T) and dihydrotestosterone (DHT) levels; penile tissues were taken for scanning electron microscopy. Results: The T, free T and DHT levels in Group B and the DHT level in Group C were significantly lower than those in Group A (P<0.05). The tunica albuginea was significantly thinner in Group B than that in Group A (P<0.05), but there was no significant difference between Group C and Group A (P>0.05). Elastic fibers in the tunica albuginea of Group A were very rich and arranged regularly and undulatedly, but in Group B, most of the elastic fibers were replaced by collagenous fibers. In Group C, the tunica albuginea was mainly composed of thick and irregular-arranged collagenous fibers. In Group A, there were abundant smooth muscle fibers in the trabeculae of corpus cavernosum, but they were much less in Group C and scarce or even disappeared in Group B. In Groups B and C, the diminished/disappeared smooth muscle fibers were replaced by irregularly arranged collagenous fibers. Conclusion: In rats, androgen is essential for maintaining the normal structure of penile tunica albuginea and corpus carvenosum.

Changes of gut flora and endotoxin in rats with D-galactosamine-induced acute liver failure

AIM: To investigate the changes of gut microflora and endotoxin levels in rats with acute liver failure (ALF) induced by D-galactosamine (GaiN).METHODS: Flora and endotoxin levels in the jejunum, ileum and colon in normal rats (group A) and rats with GaIN-induced ALF were determined at 24 h (group B) or 48 h (group C) after GaIN injection, as well as the endotoxin level in portal venous blood (PVB) and right ventricle blood (RVB) were determined by chromogenic limulus amoebocyte assay.RESULTS: Intestinal(jejunum, ileum, colon) lactobacillus count was statistically reduced in group B compared with those in group A (3.4±0.3 vs 4.9±0.3, 6.1±0.4 vs 8.0±0.3,8.1±0.2 vs 9.3±0.2, P＜0.001, P＜0.001 and P＜0.001 respectively) and recovered partially in the group C compared with those in the group B, whereas the count of Enterobacteriaceae in the jejunum, ileum and colon in group B was increased markedly compared with those in the group A (5.1±20.3 vs 3.6±0.2, 6.9±0.5 vs 5.3±0.3,8.7±0.2 vs 7.6±0.1，P＜0.001, P＜0.05 and P＜0.05 respectively)and restored partially in the group C compared with those in the group B. The endotoxin level in ileum was increased in the group B compared with those in the group A (111.3±22.8 vs 51.5±8.9, P＜0.05). In addition, the endotoxin level in PVB was obviously increased in group B compared with that in the group A (76.8±9.1 vs 40.6±7.3,P＜0.01) and reduced to the baseline at 48 h (group C).CONCLUSION: Severely disturbed gut flora in rats with GaiN-induced acute liver failure plays an important role in the elevation of endotoxin level in PVB.

Effect of compound rhodiola sachalinensis A Bor on CCI4-induced liver fibrosis in rats and its probable molecular mechanisms

AIM: To explore the anti-fibrotic effect of a traditional Chinese medicine, compound rhodiola sachalinensis A Bor on CCl4-induced liver fibrosis in rats and its probable molecular mechanisms. METHODS: Ninety healthy male SD rats were randomly divided into three groups: normal group (n=-10), treatment group of compound rhodiola sachalinensis A Bor (n=-40) and CCl4-induced model group (n=40). The liver fibrosis was induced by CCl4 subcutaneous injection. Treatment group was administered with compound rhodiola sachalinensis A Bor (0.5 g/kg) once a day at the same time. Then the activities of several serum fibrosis-associated enzymes: alanine aminotransferase (ALT), aspartate aminotransferase (AST), N-acetyl-beta-D-glucosaminidase (β-NAG) and the levels ofserum procollagen Ⅲ (PCⅢ), collagen Ⅳ (CⅣ), hyaluronic acid （HA） were assayed. The histooathol(mical chanaes were observed with HE, VG and Masson stain. The expression of TGF-β1 mRNA,αl (I) mRNA and Na+/Ca2+ exchanger (NCX ) mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) in situ.RESULTS: Compound rhodiola sachalinensis A Bor significantly reduced serum activities of ALT, AST, β-NAG and decreased the levels of PCⅢ, CⅣ, HA, improved the liver histopathological changes, inhibited the expression of TGF-β1 mRNA, α(1) mRNA and Na+/Ca2+ exchanger mRNA in rats. CONCLUSION: Compound rhodiola sachalinensis A Bor can intervene in CCI4-induced liver fibrosis in rats, in which potential mechanisms may be decreasing the production of TGF-β1, reducing the production of collagen, preventing the activation of hepatic stellate cell (HSC) and inhibiting theexpression of TGF-β1 mRNA, αl(I) mRNA and Na+/Ca2+ exchanger mRNA.

Variations of mitochondrial D-loop region plus downstream gene 1 2S rRNA-tRNA^(phe) and gastric carcinomas

AIM: To explore the instabilities, polymorphisms and other variations of mitochondrial D-loop region and downstream gene 12S rRNA-tRNAPhe in gastric cancers, and to study their relationship with gastric cancer.METHODS: Three adjacent regions (D-loop, tRNAphe and 12S rRNA) were detected for instabilities, polymorphisms and other variations via PCR amplification followed by direct DNA sequencing in 22 matched gastric cancerous tissues and para-cancerous normal tissues.RESULTS: PolyC or (CA)n instabilities were detected in 13/22(59.1％) gastric cancers and 9/22(40.9 ％) in the control (P＞0.05). There existed 2/12(16.7％) and 6/10(60％)alterations of 12S rR NA-tRNAphe in well differentiated gastric cancers and poorly differentiated ones, respectively(P0.05).Some new variations were found, among which np 318 and np 321 C-T transitions in D-loop region were two of the five bases for H-strand replication primer. Np 523 AC-deletion and np 527 C-T transition occurred at mtTF1 binding site (mtTFBS), which were associated with the transcription of downstream mitochondrial genome. Seven samples showed the np 16 182 polyC instabilities, five of which simultaneously showed np 16 189 T-C transitions.CONCLUSION: There is no statistic significance of instabilities and polymorphisms in mitochondrial D-loop region between gastric cancerous and para-cancerous normal tissues, which suggests that the instability might relate to heredity or be dependent on aging. There is asignificant correlation between differentiation degree of gastric cancer and variant frequencies of 12S rRNA-tRNAphe. The poorly differentiated gastric cancers are more prone to 12S rRNAtRNAphe variations, or gastric cancers with 12S rRNA-tRNAphe variations are more likely to be poorly differentiated, np 16189 T-C transition may be one of the important reasons for polyC instability in gastric cancer.

Tamoxifen can reverse multidrug resistance of colorectal carcinoma in vivo

AIM: To investigate the effect of tamoxifen (TAM) on multidrug resistance (MDR) of colorectal carcinoma in vivo and its relationship with estrogen receptor (ER).METHODS: Multidrug resistance was determined by means of semi-quantitative retro-transcription polymerase chain reaction (RT-PCR) to test mdr1 gene mRNA and ER expression was studied by immunohistochemistry. Tumor tissues from three cases of human colon carcinoma, which had mdr1( + )/ER(+ ), mdr1( + )/ER(-), mdr1(-) expressions,were planted subcutaneously in the neck of nude mice to establish three xenograft models. These models were subdivided into four subgroups randomly: Doxorubicin(DOX)-treated group, TAN-treated group, DOX and TAM group and control group. The dimensions of these xenografts were measured after each course of treatment and the xenografts were removed at the end of the experiments for measurements of weight and the variation of mdr1 mRNA level with RT-PCR. In each course, TAM[15 mg/(kg/d)] was administrated orally per day in the first seven days and DOX (3.6 mg/kg) was injected peritoneally on the first day. Data was evaluated by q and ttests.RESULTS: In the animal models with mdr1(-) tumor, the weights and volumes of the planted tumor in DOX group[(39.1±2.29) mg, (31.44±1.61) mm3] and TAM and DOX group [(38.72±2.56) mg, (31.31±1.74) mm3], which were lesser than that of control group [(45.48±3.92) mg,(36.42±2.77) mm3, P = 0.037, P = 0.016 respectively]significantly. In the animal models with mdr1(+)/ER(+)tumor, the weights and volumes of planted tumor were not affected by DOX or TAM treatment; however, in TAM and DOX group [(425.5±28.58) mg, (340.35±22.28) mm3],they were significantly less than that of control group[(634.23±119.41) mg, (507.45±93.34) mm3, P = 0.022,P = 0.045 respectively], which are similar to that in the models with mdr1(+)/ER(-) tumor. No significant changes were found in the expressive level of mdr1 mRNA following these treatments.CONCLUSION: The expression of mdr1 gene corresponds to the sensitivity of colon cancer to anti-tumor drugs in vivo.TAM can reverse the MDR of colorectal carcinoma in nude mice, which is independent of the expression of ER;however, no change was observed in the expressive level of mdr1 mRNA.

Frequencies of the expression of main protein antigens from Helicobacter pylori isolates and production of specific serum antibodies in infected patients

AIM: To investigate the frequencies of the expression of main protein antigens of Helicobacter pylori(H pylori)isolates, such as UreB, VacA, CagA1, HpaA, NapA, FlaA and FlaB and the production of specific antibodies in sera from H pylori-infected patients, and to understand the correlations among the different clinical types of chronic gastritis and peptic ulcer and the infection and virulence of H pylori.METHODS: H pylori strains in biopsy specimens from 157patients with chronic gastritis and peptic ulcer were isolated and serum samples from the patients were also collected.The target recombinant proteins rUreB, rVacA, rCagA1,rHpaA, rNapA, rFlaA and rFlaB expressed by the prokaryotic expression systems constructed in our previous studies were collected through Ni-NTA affinity chromatography.Rabbit antisera against rUreB, rVacA, rCagA1, rHpaA,rNapA, rFlaA and rFlaB were prepared by using routine subcutaneous immunization. By using ultrasonic lysates of the isolates as coated antigens, and the self-prepared rabbit antisera as the first antibodies and commercial HRP-labeling sheep anti-rabbit IgG as the second antibody,expression frequencies of the seven antigens in the isolates were detected by ELISA. Another ELISA was established to detect antibodies against the seven antigens in sera of the patients by using the corresponding recombinant proteins as coated antigens, and the sera as the first antibody and HRP-labeling sheep anti-human IgG as the second antibody respectively. Correlations among the different clinical types of chronic gastritis and peptic ulcer and the infection and virulence of H pylori were statistically analysed.RESULTS: In the 125 isolates of H pylori, the positive rates of UreB, VacA, CagA1, HpaA, NapA, FlaA and Flab were 100%, 65.6%, 92.8%, 100%, 93.6%, 100% and 99.2%respectively. In the 125 serum samples from the H pyloriinfected patients, the positive rates of antibodies against recombinant UreB, VacA, CagA1, HpaA, NapA, FlaA and Flab were 100%, 42.4%, 89.6%, 81.6%, 93.6%, 98.4%and 92.8% respectively. H pylori strains were isolated from 79.6% (125/157) of the biopsy specimens, but no close correlations among the H pylori infection frequencies and different types of chronic gastritis and peptic ulcer could be found (P＞0.05, x2 = 0.01-0.87). The VacA positive rate (82.40%) in the strains isolated from the specimens of patients with peptic ulcer and the anti-VacA positive rate (54.3%) in the sera from the patients were significantly higher than those (51.5%, 32.3%) from the patients with chronic gastritis (P＜0.01, x2 = 13.19; P＜0.05, x2 = 6.13).When analysis was performed in the different types of chronic gastritis, the VacA in the strains isolated from the specimems of patients with active gastritis showed a higher expression frequency (90.0%) than those from superficial (47.9%) and atrophic gastritis (30.0%) (P＜0.05, x2 = 5.93;P＜0.01,x2 = 7.50). While analysis was carried out in the strains isolated from the specimens with superficial (93.8%) and active gastritis (100%), NapA showed a higher expression frequency compared to that from atrophic gastritis (60.0%) (P＜0.01, x2 = 8.88; P＜0.05,x2= 5.00).CONCLUSION: The types of chronic gastritis and peptic ulcer and their severity are not associated with H pylori infection frequency but closely related to the infection frequency of different virulent H pylori strains. The optimal antigens for developing vaccine and diagnostic kit are UreB,FlaA, HpaA, FlaB, NapA and CagA1, but not VacA.

Liver fibrosis in chronic viral hepatitis:An ultrasonographic study

AIM: To select valuable ultrasonographic predictors for the evaluation of hepatic inflammation and fibrosis degree in chronic hepatitis, and to study the value of ultrasonography in the evaluation of liver fibrosis and compensated liver cirrhosis in comparison with serology and histology.METHODS: Forty-four ultrasonographic variables were analyzed and screened using color Doppler ultrasound system in 225 patients with chronic viral hepatitis and compensated liver cirrhosis. The valuable ultrasonographic predictors were selected on the basis of a comparison with histopathological findings. The value of ultrasonography and serology in the evaluation of liver fibrosis degree and the diagnosis of compensated liver cirrhosis was also studied and compared. Meanwhile, the influencing factors on ultrasonographic diagnosis of compensated liver cirrhosis were also analyzed.RESULTS: By statistical analysis, the maximum velocity of portal vein and the degree of gall-bladder wall smoothness were selected as the valuable predictors for the inflammation grade (G), while liver surface, hepatic parenchymal echo pattern, and the wall thickness of gall-bladder were selected as the valuable predictors for the fibrosis stage (S). Three S-related independent ultrasonographyic predictors and three routine serum fibrosis markers (HA, HPCIII and CIV) were used to discriminate variables for the comparison of ultrasonography with serology. The diagnostic accuracy of ultrasonography in moderate fibrosis was higher than that of serology (P<0.01), while there were no significant differences in the general diagnostic accuracy of fibrosis as well as between mild and severe fibrosis (P<0.05). There were no significant differences between ultrasonography and serology in the diagnosis of compensated liver cirrhosis.However, the diagnostic accuracy of ultrasonography was higher in inactive liver cirrhosis and lower in active cirrhosis than that of serology (both P<0.05). False positive and false negative results where found when the diagnosis of compensated liver cirrhosis was made by ultrasonography.CONCLUSION: There are different ultrasonographic predictors for the evaluation of hepatic inflammation grade and fibrosis stage of chronic hepatitis. Both ultrasonography and serology have their own advantages and disadvantages in the evaluation of liver fibrosis and compensated liver cirrhosis. Combined application of the two methods is hopeful to improve the diagnostic accuracy.

Inhibition of human La protein by RNA interference downregulates hepatitis B virus mRNA in 2.2.15 cells

AIM: To investigate the role of human La protein in HBV mRNA expression.METHODS: Three human La protein (hLa) specific siRNA expression cassettes (SECs) containing U6+1 promoter were prepared via one-step overlapping extension PCR. After transfection with SECs into HepG2 cells, inhibition effects on hLa expression were analyzed by semi-quantitative RT-PCR and Western blotting. Then, effective SECs were screened out and transfected into 2.2.15 cells, a stable HBV-producing cell line. HBV surface antigen(HBsAg) and e antigen (HBeAg) secretions into culture media were detected by microparticle enzyme immunoassay (MEIA) and HBs and HBe mRNA levels were analyzed by semi-quantitative RT-PCR.RESULTS: SEC products containing U6+1 snRNA promoter,and 3 sites of hLa mRNA specific siRNA were obtained successfully by one-step overlapping extension PCR and could be directly transfected into HepG2 cells, resulting in inhibition of La protein expression in both mRNA and protein levels, among which U6+l-hLa833 was the most efficient,which reduced 18.6-fold mRNA and 89% protein level respectively. In 2.2.15 cells, U6+l-hLa833 was also efficient on inhibition of hLa expression. Furthermore, semi-quantitative RT-PCR showed that HI3s and HBe mRNA levels were significantly decreased by 8-and 66-fold in U6+l-hLa833 transfected cells compared to control. Accordingly, HBsAg and HBeAg secretions were decreased partly posttransfection with SECs.CONCLUSION: PCR-based SECs can be used to mediate RNAi in mammalian cells and provide a novel approach to study the function of La protein. The inhibition of La protein expression can result in a significant decrease ofHBV mRNA, which implies that the hLa protein is also involved HBV RNA metabolism as one of the HBV RNA-stabilizing factors in human cells.

Ultrastructural changes of penile tunica albuginea in diabetic rats

Aim: To clarify the ultrastructural changes of penile tunica albuginea (TA) in streptozotocin (STZ)-induced diabetic rats. Methods: Intraperitoneal injection of STZ was used to induce diabetes mellitus (DM) in 12 Sprague Dawley rats. Ten rats (age and weight-matched) were used as control. Blood samples from the tail snips of the rats were used for the determination of serum glucose levels with SureStep Plus Blood Meter. At week 4 and 10 after the injection, half of the rats in each group were sacrificed and penile samples were obtained from the middle third of the penile shaft for the examination of TA under scanning electron microscopy. Results: In the diabetic group, the serum glucose levels were higher (P<0.01 at both time points) and the TA were thinner (P<0.05) than those of the controls. In the control group, the fibers of TA were rich and arranged regularly and undulated, while in the diabetic group, the fibers were diminished, lost the undulations and were arranged irregularly. Conclusion: In rats, DM appeared to impair the penile TA ultrastructures and this impairment could contribute to diabetic erectile dysfunction in part by impairing the veno-occlusive function.

Effects of atorvastatin on vascular remodeling in spontaneously hypertensive rats

Objective: To investigate the structural changes of aorta, and evaluate the effects of atorvastatinon the remodeling of thoracic aorta in spontaneously hypertensive rats(SHR) . Methods : Twelve eight-week-old SHR were randomized into atorvastatin treated group( ATV group, n = 6) and distilled water group( DW group, n = 6) ; Wistar-Kyoto rats(WKY) were used as normal controls. Atorvastatin was administered to ATV group for 10 weeks by gavage in mixture with distilled water( 1ml) ; the latter two groups were given the same amount of distilled water by gavage for 10 weeks. Systolic blood pressure of caudal artery was examined before and after treatment, and serum concentrations of total cholesterol, triglycerides and HDL-C were measured.Wall thickness, media thickness, medial cross-sectional area and lumen diameter of thoracic aorta were assessed with computed video processing. Results: Systolic blood pressure in ATV group was markedly lower than that in DW group( P < 0.01). Compared with DW group and WKY group, serum concentrations of total cholesterol, triglycerides and HDL-C in ATV group were significantly lower( P < 0.01, P < 0.05). Wall thickness, media thickness, and medial cross-sectional area to lumen ratio in DW group were significantly higher than those in WKY group and ATV group( P < 0.01, P < 0.05), but no such difference was found between WKY group and ATV group( P > 0.05 ). Conclusion : Vascular structural changes of aorta are due to the alteration of the vessel wall in early stage of SHR. Atorvastatin can markedly improve vascular remodeling.

Cytomegalovirus and chronic allograft rejection in liver transplantation

Cytomegalovirus (CMV) remains one of the most frequent viral infections and the most common cause of death after liver transplantation (LT). Chronic allograft liver rejection remains the major obstacle to long-term allograft survival and CMV infection is one of the suggested risk factors for chronic allograft rejection. The precise relationship between cytomegalovirus and chronic rejection remains uncertain.This review addresses the morbidity of cytomegalovirus infection and the risk factors associated with it, the relationship between cytomegalovirus and chronic allograft liver rejection and the potential mechanisms of it.

Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro

AIM: To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA)expression vector.METHODS: Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vector and shRNA expression vectors were constructed and cotransfected transiently into HepG2 cells. mRNAs extracted from HepG2 cells were detected by real-time PCR. Fluorescence of HBs-GFP protein was detected by fluorescence-activated cell sorting (FACS). The effective shRNA expression vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in HepG2.2.15 cells were analyzed by radioimmunoassay (RIA) method.RESULTS: FACS revealed that shRNA targeting at HBsAg reduced the GFP signal by 56% compared to the control.Real-time PCR showed that HBs-GFP mRNA extracted from HepG2 cells cotransfected with pAVU6+27 and HBs-GFP expression plasmids decreased by 90% compared to the empty vector control. The expressions of HBsAg and HBeAg were also inhibited by 43% and 64%, respectively.CONCLUSION: RNAi using shRNA expression vector can inhibit the expression of HBsAg, providing a fresh approach to screening the efficient small interfering RNAs (siRNAs).

Genomic determination of CR1 CD35 density polymorphism on erythrocytes of patients with gallbladder carcinoma

AIM: To study the changes of quantitative expression, adhering activity and genomic density polymorphism of complement types in erythrocytes (CR1) of patients with gallbladder carcinoma and the related clinical significance.METHODS: Polymerase chain reaction (PCR), HindⅢ restrdion enzyme digestion, quantitative assay of CR1 and adhering activity assay of CR1 in erythrocytes were used.RESULIFS: The number and adhering actvity of CR1 in patients with gallbladder carcinoma (0.738±0.23, 45.9±5.7) were significantly lower than those in chronic cholecystitis and cholecystolithiasis (1.078±0.21, 55.1±5.9) and healthy controls (1.252±0.31, 64.2±7.4) (P<0.01). The number and adhering activity of CR1 in patients with chronic cholecystitis and cholecystolithiasis (1.078±0.21, 55.1±5.9) were significantly lower than those in healthy controls (1.252±0.31, 64.2±7.4) (P<0.05). There was a positive correlation between quantitative expression and adhering activity of CR1 (r = 0.79, P<0.01).Compared with those on preoperative day (0.738±0.23,45.4±4.9), the number and adhering activity of CR1 in patients with gallbladder carcinoma decreased greatly on the third postoperative day (0.310±0.25, 31.8±5.1) (P<0.01), and on the first postoperative week (0.480±0.25, 38.9±5.2) (P<0.01), but they were increased slightly than those on the preoperative day (P>0.05). The number and adhering activity of CR1 recovered in the second postoperative week (0.740±0.24, 46.8±5.9) (P<0.01) and increased greatly in the third postoperative week (0.858±0.35, 52.7±5.8) (P<0.01) in comparison with those on the preoperative day and in the first postoperative week. The number and adhering activity of CR1 of gallbladder carcinoma patients with infiltrating, adjacent lymphogenous and distant organ metastases were significantly lower than those of gallbladder carcinoma patients without them (P<0.01). No difference was observed between the patients with gallbladder carcinoma and healthy individuals in the spot mutation rate of CR1 density gene (%2 = 0.521, P>0.05).The distribution of expression was 67.8% in high expression genomic type, 24.8% in moderate expression genomic type,and 7.4% in low expression genomic type. The number and adhering activity of CR1 high expression genomic type gallbladder carcinomas (0.749±0.22, 42.1±6.2) were significantly lower than those of healthy individuals(1.240±0.29, 63.9±7.2), and were also significantly lower than those of healthy individuals (0.921±0.23, 54.8±7.1), but no difference was observed between the number and adhering activity of CR1 lower expression genomic type gallbladder carcinomas (0.582±0.18, 44.3*_5.5) and those of healthy individuals (0.610±0.20, 45.8±5.7) (P>0.05).CONCLUSION: Defective expression of CR1 in gallbladder carcinoma is mostly acquired through central peripheral mechanisms. The changes in CR1 quantitative expression and adhering activity are consanguineously related to the development and metastasis in gallbladder carcinoma.

Inducible nitric oxide synthase, nitrotyrosine and apoptosis in gastric adenocarcinomas and their correlation with a poor survival

AIM: To detect the presence of inducible nitric oxide synthase (iNOS), nitrotyrosine (NT) and apoptosis in gastric adenocarcinomas and their possible correlations with the clinicopathological characteristics and prognosis of gastric adenocarcinoma.METHODS: Sixty-six specimens of gastric adenocarcinoma and corresponding adjacent normal gastric tissues were studied. Immunohistochemistry was employed to localize iNOS and NT protein and an immunohistochemical scoring system was used. The occurrence of apoptotic cell death (apoptotic index [AI]) was analyzed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labeling (TUNEL) method. RESULTS: Results showed that iNOS expression wasdetected at an intermediate or high level in 41 of 66 (62%) specimens of gastric adenocarcinoma. NT expression was 58%. Neither of them was found in the normal gastric tissues; there were significant positive correlations among iNOSexpression, NT expression and AI. Many clinicopathologic characteristics of gastric adenocarcinoma, such as tumor size, depth of invasion, lymph node metastasis and TNM staging, were related to iNOS and NT expressions (P＜0.05). In 66 surviving patients, the 5-year survival rate of 41 patients who had tumors with intermediate or high iNOS expressions and high Ais (4.09%; 19.96%) was significantly lower than that of 25 patients who had tumors with negative or low iNOS expressions and low Ais (0.79%; 47.14%) (P= 0.001). COX's multivariate analysis revealed that the iNOS expression was identified as one of the significant independent prognostic factors predictive of a poor survival (relative risk [RR] = 2.69).CONCLUSION: NO produced by iNOS may play a stronger role in promoting gastric adenocarcinoma growth than in suppressing its growth. iNOS and NT expressions by gastric adenocarcinoma may correlate with a poor survival.

Expression of platelet-derived growth factor-BB in liver tissues of patients with chronic hepatitis B

AIM:To study the relationship between expression of plateletderived growth factor-BB (PDGF-BB) and fibrogenesis in chronic hepatitis B.METHODS:Hepatic tissues from 43 patients with chronic hepatitis B were embedded in paraffin.The sections were stained with HE and picric acid-sirius red to determine inflammatory activity and fibrosis stages. PDGF-BB expression was detected by immunohistochemistry and assessed semiquantatively. Levels of serum hyaluronic acid (HA),pro-collagen Ⅲ(PCⅢ), collagen Ⅳ (Ⅳ-C) and laminin (LN) were examined by radioimmunoassay (RIA).RESULTS:The expression level of PDGF-BB was found to be positively correlated with inflammatory activity, fibrosis stage and grade of histological findings (τ=0.58,0.55,0.55,P<0.01).The positive correlation was also observed between tissue level of PDGF-BB expression and contents of HA, PCⅢ, Ⅳ-C and LN in the circulation (r=0.52,0.32,0.40,0.33, P<0.05).CONCLUSION: PDGF-BB may play some role in the development and progression of liver fibrosis.

Cyclooxygenase-2 promotes angiogenesis by increasing vascular endothelial growth factor and predicts prognosis in gallbladder carcinoma

AIM: To investigate the relationships between theexpression of cydooxygenase-2 (COX-2), vascular endothelial growth factor (VEGF) and the degree of vascularization, clinicopathologic feature, survival time of patients with gallbladder carcinomas.METHODS: Sixty-four gallbladder carcinoma specimens were evaluated for COX-2, VEGF expression by immunohistochemical methods. Microvessel counts (MVC) were determined using CD34. The relationships between COX-2,VEGF expression, CD34-stained MVC, clinicopathologic features and survival time were analyzed. The correlations between COX-2 and VEGF expression, CD34-stained MVC were also investigated.RESULTS: COX-2, VFGF immunoreactivity were observed in 71.9% (46/64) and 54.7% (35/64) specimens,respectively. The average MVC in 64 cases of gallbladder carcinoma was 57±14 per high power vision field. The status of MVC was closely correlated with Nevin staging, tumor differentiation and lymph node metastasis (P＜0.01,0.002, and 0.003, 0.000, respectively). Increased VEGF expression was significantly correlated with tumor differentiation (poorly and moderately＞well differentiated, P＜0.05, P = 0.016). Clinical stages had no relation with the expression of VEGF (P＞0.05, P = 0.612). There was a positive correlation between COX-2 expression and clinical stages. The positive rate of COX-2 was higher in cases of Nevin stages S4-S5 (81.8%) than in those of Nevin stages S1-S3 (50.0%) with a statistical significance (P＜0.01, P = 0.009). The expression of COX-2 did not vary with differentiation (P＞0.05, P = 0.067). Statistically significant differences were also observed according to lymph node metastasis, COX-2 expression and VEGF expression(P＜0.01, 0.000, and 0.001, respectively). There was no relation between VEGF, COX-2 expression, MVC and the age and sex of patients. MVC and VEGF positive rate in the COX-2 positive gallbladder carcinoma tissue was higher than that in the COX-2 negative tissue (P＜0.05, 0.000, and 0.032, respectively). Patients with VEGF, COX-2 positive tumors had a significantly shorter survival time than those with negative tumors (P＜0.05, 0.004, 0.01, respectively).CONCLUSION: Augmented tumor neovascularization induced by VEGF may be one of the several effects of COX-2 responsible for poor prognosis of human gallbladder carcinoma. COX-2 inhibitor, either in combination therapy with other agents, or for chemoprevention, may be effective via suppression of angiogenesis in this fatal disease.

Study of clinical features of amyloid angiopathy hemorrhage and hypertensive intracerebral hemorrhage

Objective: The purpose of this study was to differentiate between cerebral amyloid angiopathy (CAA) and hypertension (HTN) based on hemorrhage pattern interpretation. Methods: From June 1994 to Oct., 2000, 83 patients admitted to our service with acute intracerebral hemorrhage (ICH) were investigated retrospectively; 41 patients with histologically proven diagnosis of cerebral amyloid angiography and 42 patients with clear history of hypertension were investigated. Results: Patients with a CAA-related ICH were significantly older than patients with a HTN-related ICH (74.0 years vs 66.5 years, P<0.05). There was a significantly higher number of hematomas≥30ml in CAA (85.3%) when compared with HTN (59.5%). No basal ganglional hemorrhage was seen in CAA, but in 40.5% in HTN. In CAA-related ICH, subarachnoid hemorrhage (SAH) was seen in 26 patients (63.4%) compared to only 11 patients (26.2%) in HTN-related ICH. Intraventricular hemorrhage was seen in 24.4% in CAA, and in 26.2% in HTN. Typical features of CAA-related ICH included lobar distribution affecting mainly the lobar superficial areas, lobulated appearance, rupture into the subarachnoid space, and secondary IVH from the lobar hemorrhage. More specifically, multiplicity of hemorrhage, bilaterality, and repeated episodes also strongly suggest the diagnosis of CAA. Multiple hemorrhages, defined as 2 or more separate hematomas in multiple lobes, accounted for 17.1% in CAA-related ICH. Conclusion: There are certain features in CAA on CT and MRI and in clinical settings. To some extent, these features may contribute to distinguishing CAA from HTN related ICH.

Effect of endotoxin on portal hemodynamic in rats

AIM: To study the effects of endotoxin on portalhemodynamic of normal and noncirrhotic portal hypertensiverats.METHODS: Normal rats were intraperitonealy injected with 0.1,0.25, 0.5, 1.0, 2.0, 4.0 mg@kg-1 of lipopolysaccharide( LPS ) respectively, portal vein ligation ( PVL ) andintrahepatic portal occlusion (IPO) rats as well as sham-operated rats were treated with an intraperitoneal injection of1.0 mg@kg-1 of LPS, the portal vein pressure(PVP), portalvenous flow(PVF), inferior vena cava pressure(IVCP) andportal vein resistance (PVR) were detected 4 hours afterinjection.RESULTS: PVF of the 5 groups of rats acceptingintraperitoneal injection of LPS were increased from 14.0 to18.0, 22.2, 266.2, 34.8, 39.66, 38.8 mL@min-1 4 hours afterinjection of LPS ( P ＜ 0. 01 ). PVP of the 4 groups of ratsaccepting more than 0.1 mg/kg@ b. w of LPS was increasedfrom 1.04 to1.25, 1.50, 1.80, 1.95, 2.05 kPa(P＜0.01). Theincrements of PVF and PVP were in a dose-dependentmanner of LPS. PVR of the 5 groups of rats was decreasedfrom 51 to 42,44,48,45,44,47 kPa@min@ L-1 ( P＜ 0.05) and nodosedependent manner wes observed. PVF of PVL, IPOand sham-operated rats increased from 22.66 to 32.8, 22.0 to28.0, 14.0 to 34.8 mL@min1 ( P＜ 0.01), and PVP increasedfrom 1.86 to 2.24, 1.74 to 1.95, 1.04 to 1.80 kPa(P＜0.01),PVR decreased from 71 to61, 67 to61, 52 to 44 kPa@min@ L-1after intraperitoneal injection of 1 mg@ kg-1 of LPS. Theincrements of PVF and PVP of PVL and IPO rats weresignificantly less than the sham-operated rats ( P ＜ 0. 01 ),There wes no significant difference between the amounts ofPVR decreased in the two groups of PHT model rats andsham-operated rats( P＞ 0.05) after intraperitoneal injection 1mg@kg1 of LPS.CONCLUSION: Endotoxin could prompt portal hypertensionof the normal and noncirrhotic portal hypertensive rats byincreasing portal blood flow mainly.

Suppressing progress of pancreatitis through selective inhibition of NF-κB activation by using NAC

Objective:To explore the characteristics of NF-κB activation in the progress of pancreatitis，the relationship with expression of TNF-α in the inflammatory reaction, and prevent the exacerbation of pancreatitis by using NAC.Method:Forty-eight rats were divided into three groups: therapy (group C), pancreatitis (group B) and control (group A).NAC served as the inhibitor of NF-κB activation.In the time intervals of 1.5, 3.0, 6.0, 12.0 hour, NF-κB activation was detected with flow cytometry (FCM) and the expression of TNF-α mRNA and protein with in situ hybridization (ISH) and enzyme-linked immuno-sorbent assay (ELISA) respectively. Meanwhile, the level of lipase and amylase in the serum was assayed and the pathological change was evaluated. Result:NF-κB activation in the pancreatitis group was higher than that in the control group (P<0.01),peaked at 3 hours,and was depressed by the inhibitor of NF-κB, NAC.The expression of TNF-α as well as the level of lipase and amylase in the serum also rose synchronously with activation of NF-κB.In contrast to group A, it was significantly different (P<0.01) in group B.After using NAC in group C,all of these values were decreased and the inflammatory reaction in the pancreas abated evidently. The pathology changes of the pancreas were shown to be alleviated in group C. Conclusion: First,NF-κB activity is intensively initiated in the course of pancreatitis and shown to have closely relationship with the release of cytokines. Second, use of NAC markedly depressed NF-κB activation. TNF-α expression is down regulated by cytokines. It is suggested that NAC probably acts as a useful agent for treatment of pancreatitis by indirectly inhibiting activation of NF-κB.