Volume 89  ·   Issue 12  ·  2009

Colorectal cancer is one of the common malignant tumors. 60% ~70% of colorectal cancer patients will have liver metastasis, of which about 25% have synchronous liver metastasis and 35% ~45% have metachronous liver metastasis. The median survival time of untreated patients is 6~18 months [1].

Objective To investigate the sustained release rule of doxorubicin/polylactide-grafted dextran copolymer (DOX/DEX-PLA) nanoparticles and the effect thereof in killing hepatocarcinoma cells. Methods DOX/DEX-PLA nanoparticles were prepared by method of emulsification & evaporation of organic solvent. Its morphology was observed by transmission electron microscopy and the encapsulating efficiency of DOX was determined by ultraviolet spectrophotometry. DOX/DEX-PLA was put in a dialysis bag to observe the releasing characteristics of DOX from DOX/DEX-PLA nanoparticles in vitro. Human liver carcinoma cells of the fine HepG2 were cultured with DOX/DEX-PLA of different concentrations or the original drug of DOX as control group for 24, 36, or 48 h. MTT method was used to observe the cancer inhibition rate. BALB/c nude mice underwent subcutaneous injection of HepG2 cells at the right scapula and then randomly divided into 4 groups to undergo intravenous injection of DOX/DEX-PLA (excremental group), original drug of DOX (naked drug group), DEX-PLA (nanovector group), or normal saline (blank control group) once every 5 days for 3 times. Twenty-one days later the mice were killed with the tumors taken out to measure the weight to analyze the inhibitory effect against hepatocarcinoma cells. Results The DOX/DEX-PLA nanoparticles were of round or elliptical shape with the diameter of about 83 nm, and the DOX entrapment efficiency was about 67. 1%. The releasing test in vitro manifested a sustained release of over 50% of DOX encapsulated in DOX/DEX-PLA nanoparticles for about 7 days. Both the DOX/DEX-PLA nanoparticles and naked drug DOX inhibited the growth of HepG2 cells with a similar inhibitory rate (51.3% vs 50. 7%, P >0. 05), meanwhile the DEX-PLA nanovector failed to inhibit the HepG2 cells. In-vivo experiment showed an inhibitory rate of DOX/DEX-PLA nanoparticles on hepatocellular carcinoma xenografts of 68.56%, significantly higher than that of the naked drug DOX (48. 17%). Conclusion DOX/DEX-PLA nanoparticles can effectively inhibit the growth of hepatocarcinoma cells.

Objective To evaluate the efficacy and safety of transcatheter arterial chemoembolization combined with percutaneous ethanol injection in treating unresectable primary liver cancer. Methods MEDLINE (1966 - June 2008), EMBASE (1974 - June 2008), the Cochrane Library (Issue 2, 2008), CBMdisc (1978-June 2008), CNKI (1979-June 2008), and Wanfang Database were searched without language limitation. All randomized controlled trials comparing transcatheter arterial chemcembolization alone with transcatheter arterial chemoembolization combined with percutaneous ethanol injection for treating unresectable primary liver cancer were identified and screened by two reviewers, and the methodological quality of the included randomized controlled trials were evaluated by Jadad scale. The Cochrane Collaboration's RevMan4. 2 software was used for the analysis of the data extracted from the included randomized controlled trials. Results Fourteen randomized controlled trials involving 857 patients were included with 12 trials published in Chinese and 2 in English. Meta-analysis of the data extracted from the included randomized controlled trials showed that transcatheter arterial chemoembolization combined with percutaneous ethanol injection could significantly improve the overall survival rates compared with transcatheter arterial chemoembolization alone , with the corresponding relative risk (RR) values (95% CI) for the 1,2, and 3-year survival of 1.37 (1.21 - 1.56), 1.74 (1.49 -2. 04), and 2. 26 (1.70 -3. 02) respectively; and the RR alpha-feW-protein (AFP) negative conversion rate, AFP lowering rate, and tumor focus efficacy rate were 1.39 (1.24 - 1.56), 1.69(1.38 -2. 07), and 1.56 (1.38 - 1.77) respectively. Side effects or adverse events related with transcatheter arterial chemoembolization were reported in 11 randomized controlled trials, mainly liver function impairment, fever , gastrointestinal symptom, and transient pain, and no major treatment-related complication or death was reported. Conclusion Compared with transcatheter arterial chemoembolization alone, transcatheter arterial chemoembolization combined with percutaneous ethanol injection significantly is a better approach in treating unresectable primary liver cancer, increasing the overall survival rate, AFP negative and lowering rates, and tumor response rate. However, the methodological quality of most reported randomized controlled trials is low.

Objective To investigate the effects of nanogold in inhibition of angiogenesis and growth of liver cancer cells. Methods Nanogold was co-incubated with VEGF165 and VDGF121 respectively. Atomic forces microscopy (AFM) was used to observe the changes of the form of the particles. Human umbilical vascular endothelial cells (HUVEC) were serum-starved for 24 hours, then co-cultured with VEGF165 + nanogold or VEGF121 + nanogold for 24 h. ATM was used to observe the ultrastructure of the cells. Another HUVEC were serum-starved for 24 hours and then cultured with VEGF165 (10 μg/L) 100 μl + nanogold 125,250,and 500 nmol/L 100 μl respectively for5 min. Then Western blotting was used to detect the phosphorylation protein of phospholipase C (PLC)γ1 on VEGFR-2. Hepatocellular cancer ceils of the line H22 were injected subcutaneously into the fight armpits of 20 Balb/c nude mice. When the size of transplanted tumor reached about 8 mm, the mice were divided into 2 equal groups: experimental group undergoing injection of nanogold into the tumor once a day for 8 days, and control group injected with normal saline. On day 14 the mice were sacrificed with the liver tumors taken out to measure the size and weight. The microvascular density (MVD) of tumor was determined by immunohistochemical staining.Results ATM showed that acted with VEGF165, the size of nanogold became over 30 nm. Treated with VEGF165 the HUVEC became larger with obvious pseudopodiums. However, such changes were obviously milder in those HUVEC treated with nanogold + VEGF165. The PLC-γ1 phosphorylation level VEGF receoptor-2 was decreased along with the increase of the concentration of nanogold. The MVD of liver cancer tissue in the experimental group was 14. 27 ± 1.08, significandy lower than that of the control group [(23.52 ± 1.36), P <0. 01]. The mean weight and volume of tumor of the experimental group were (1.39 ± 0. 08) g and (1.37 ± 0. 34) cm3 respectively, both significantly lower than those of the control group [(2. 47 ±0. 15) g and (2. 49±0. 78) cm3 respectively, both P <0. 05] with a tumor growth inhibition rate of 43.72%. Conclusion Nanogold significantly inhibits the angiogenesis and growth of liver cancer cells with the possible mechanism that nanogold inhibits the VEGF165-induced signaling.

Objective To investigate the inhibitory effect of nanoparticle-mediated endostatin gene therapy on hepatocellular carcinoma xenografts combined with local hyperthermia utilizing heat-inducible promoter. Methods Heat-inducible HSP70B promoter and fusion gene of Endo/EGFP were cloned into pcDNA3.1 (+) plasmid, thus obtaining recombinant plasmid of pcDNA3. 1 (+)/HSP70-Endo/EGFP using restriction endonucleases Bgl Ⅱ/Hind Ⅲ and EcoR Ⅰ/Sal Ⅰ. The nanoparticles polylactide-grafted dextran copolymer (DEX-g-PLA) encapsulating the recombinant plasmid DNA were prepared by the method of emulsification and evaporation of organic solvent, and the surface shape of nanoparticles was observed transmission electron microscope. Human hepatocellular cells of the lines HepG2 and ECV304 were cultured and transfected with the recombinant plasmid utilizing the nanoparticles. Following thermal induction at 37 ℃, 39 ℃, 41 ℃, 43 ℃, and 45 ℃ for 30 min, the expression of enhanced green fluorescent protein (EGFP) was detected by fluorescence microscope and flow cytometry. The concentration of endostatin protein in the supernatant was tested by ELISA, and the growth inhibition on the HepG2 and ECV304 cells was tested by MTT method. Balb/c nude mice were inoculated with HeG2 cells and then randomly divided into 2 groups to undergo intra-tumor injection of nanoparticles (heated or not heated), Lipofectamine 2000. Mice were used as controls without intra-tumor injection. Four weeks the mice were killed to observe the tumor inhibition rate. Results The nanoparticles encapsulating recombinant plasmid were of round elliptical shape 90 ～ 120 nm in diameter. The efficiency of gene transfection mediated by nanoparticles was about 30.65%. The expression of Endo/EGFP gene in the HepG2 cells was up-regulated along with the increase of temperature, peaked at 43 ℃(with the EGFP expression level 3.3 times as that at 37 ℃). The concentration of endostatin protein in the aupernatant of the 43 ℃ group was (177 ±28)μg/L, significantly higher than that of the 37 ℃ group [(41 ± 10) μg/L]. MTT results indicated that endostatin inhibited the growth of ECV304 cells with a inhibition rate of 96. 3% at the time point of 72 h in the 43 ℃ group, however, it did not show influence on HepG2 cells no matter what was the temperature The tumor inhibition rate in the mice of endostatin with thermal induction group was 58.5%, significantly higher than that of the 37 ℃ group (34.9%, P < 0. 05) .Conclusion Low temperature thermal induction enhances the expression and secretion of endostatin in hepatocellular cells transfected by nanoparticles, and inhibits the growth of hepatocellular carcinoma xenografts.

Objective To explore the role of bacteria in etiology of chronic pelvic pain syndrome (CPPS) , i.e. , chronic prostatitis and the correlation between presence of bacterial signal of 16S ribosomal RNA (16S rRNA) gene and the response to antibiotics. Methods Samples of prostatic and subcutaneous tissues were obtained by biopsy via perineal approach from 112 CPPS patients, aged 20 - 48. Polymerase chain reaction was conducted to detect the 16S rRNA gene of bacteria. The patients were treated with gatifloxacin 0. 4 g once a day for 4 weeks and then 4 weeks later the effects of treatment were assessed by the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI). Results PCR was completed in 94 of the 112 patients, and 18 were excluded because their subcutaneous biopsies were positive for 16S rRNA, showing the possible contamination of their prostatic tissues. The total positive rate of bacterial 16S rRNA gene was 63. 8% (60/94). The positive rate of bacterial 16S rRNA gene in the patients with Ⅲa CPPS and Ⅲb CPPS were 68. 3% and 60. 3% respectively. The total gatifloxacin effective rate of positive bacterial signal group after the was 55.0%, significantly higher than that of the negative bacterial signal group(14. 7%, P <0. 001). The gatifloxacin effective rate of the 16S rRNA positive Ⅲa CPPS patients was 75% , significantly higher than that of the 6S rRNA negative Ⅲa CPPS patients (23. 1%, P < 0. 001) , and the gatifloxacin effective rate of the 16S rRNA positive Ⅲb CPPS patients was 37. 5%, significantly higher than that of the 6S rRNA negative Ⅲb CPPS patients (9. 5%, P <0. 05). Conclusion Bacterial infection is related to CPPS in part of the patients. Bacterial signal detection helps predict the effect of antimicrobial therapy.

：Objective To study therelationship between tumor infiltration in mesorectum and prognosis of middle and lowerrectal cancer. Methods 49 patients with middle and lower rectal cancer underwent totalmesorectal excision. Specimens were obtained during operation and underwent large slicepathologic technique to observe the degree of tumor infiltration in mesorectum andcircumferential resection margin. Follow-up was conducted for 61 (9 - 66) months toobserve the local recurrence rate, metastasis rote, and five-year survival rate. ResultsFollow-up showed a local recurrence rate of 12. 2% (6/49), distant metastasis rate of 26.5% (13/49), and five-year survival rate of 67. 3% (33/49). The rate of degree Ⅰ of tumor infiltration in mesorectum was 40. 8% (20/49), the degreeⅡ rate was 26. 5% (13/ 49), and the degree Ⅲ rate was 32. 7% (16/49) with thecorresponding local recurrence rates of 0, 7. 7% (1/13) , and 31.3% (5/16) respectively(x2 = 7. 357, P = 0. 015), metastatic rates of 10% (2/20), 23. 1% (3/13), and 50% (8/16)respectively (x2 = 7. 405, P = 0. 025), and the 5-year survival rates of 90% (18/20), 69.2% (9/13), and 37. 5% (6/16)respectively . Kaplan-Meier survival analysis showed that thesurvival time was correlated with the degree of tumor infiltration in mesorectum (P = 0.012). The rate of circumferential resection margin involvement was 24. 5% (12/49). In the12 patients with positive circumferential resection margin, the local recurrence rate wag33. 3% (4/12), whereas 5.4% (2/37) in those with negative circumferential resection margin(x2 = 6.577, P = 0.010). Distant metastasis rate was 50% (6/12) in the patients withpositive circumferential resection margin, whereas 18. 9% (7/37) in those with negativeone (x2 =4. 491 ,P =0. 034). The 5-year survival rate of the patients with positivecircumferential resection margin was 33. 3% (4/12), significantly lower than that of thepatients with negative circumferential resection margin [78. 4% (29/37)] . Kaplan- Meiersurvival analysis showed survival time was correlated with the circumferential resectionmargin status (P = 0. 009). Conclusion The degree of tumor infiltration in mesorectum andcircumferential resection margin status are important predictors of local and distantrecurrence as well as survival of patients with middle and lower rectal carcinoma. `

Objective To investigate the percentage and significance of CD4+ CD25+ Foxp3+ regulatory T cells in the patients with heamorrhagic fever with renal syndrome (HFRS). Methods Peripheral blood samples were collected from 27 HFRS patients, and 19 healthy controls to examine the proportion of the CD4+ CD25+Foxp3 + cells. RT-PCR was used to detect the Foxp3 mRNA expression in the mononuclear cells in peripheral blood. Biopsy was conducted on the HFRS patients during oliguria stage. Peripheral blood samples and morning urine samples were collected in the stages and in the very day of renal biopsy to examine the urine protein, urine creatinine (Cr), blood Cr, and creatinine clearance rate (Ccr). Results The percentages of CD4+ CD25+ Foxp3+> in the HFRS patients in the hypetension, oliguria, polyuria, and convalescent stages were 5.0% ± 1.2%, 3. 1%± 0. 8%, 4. 2% ± 0. 9%, and 5.0% 4± 1.1% respectively, all significantly lower than that of the normal control group (6. 5% ± 2. 1%, all P < 0. 01). The proportion of CD4+ CD25+ Foxp3 +> T cells in the CD4 +> T cells in the oliguria stage was significantly lower than those in other stages and that of the normal controls (P <0. 01). The percentage of CD4 + CD25 + Foxp3 + in the patients with HFRS was significantly correlated with the renal injury index (r = 0. 51, P <0. 01), but not significantly correlated with urine protein and renal function changes (r =0. 11 and 0. 18, both P > 0. 05). The expression level of Foxp3 mRNA in peripheral mononuclear cells of the HEFS patients in the oliguria stage was (503 ± 64), significantly lower than that of the normal controls (3102 ±409, P <0. 01) and those of the HFRS patients in the hypotension, polyuria, and convalescent stages (1805 ±291, 1869 ±320, and 2697±420,all P <0.01). Conclusion The proportion of CD4+ CD25+ Foxp3 + regulatory T cells decreases in patients with HFRS, which may contribute to the pathological process in such disease.

Objective To investigate the relevant factors of seroresistance of syphilis by clinical analysis and study of immunological function. Methods The clinical data of 172 patients with syphilis were analyzed to observe the correlation of seroresistance with the factors including original titer of rapid plasma regain (RPR), disease course, and medication. Flow cytometry was used to analyze the proportions of T-lymphocyte subset and NK cells in the peripheral blood of 40 untreated syphilitic patients at various stages, 20 syphilitic patients whose serology turned negative after treatment, 20 serum resistance patients, and 18 healthy controls. Results The seroresistance rote was 8. 7% in the primary syphilis patients (2/23), significantly lower than that of the latent syphilis patients (42. 2%, 38/90, P =0. 001). The seroresistance rate of the patients treated with benzathine penicillin was 26. 4% (42/159), significantly lower that of the patients treated with microlides (76. 9%, 10/13, P = 0. 000). The proportion of CD8+ T cells of the untreated syphilitic patients was (30% ± 8%), significantly higher than that of the normal controls [(24% ±4%) ,P < 0. 01]. The proportion of NK cells of the untreated syphilitic patients was (13% ± 5%), significantly lower than that of the normal controls (16% ± 6%, P < 0. 05). The proportion of CD3+T cells of the patients whose serology turned negative after treatment was (71% ±7%), significantly higher than that of the normal controls (65%±7% ,P <0.05). The proportion of CD8+ T cells of the patients whose serology turned negative after treatment was(29%±7%), significantly higher than that of the normal controls(24% ±4% ,P <0. 01). There were not significant differences in the proportions of different cells among the patients with seroresistance, serological negative conversion, and normal controls. Conclusion Syphilis patients with a high baseline RPR titer, a latent course and macrolide therapy are prone to be seroresistant Whether the occurrence of serum resistance is related to the inhibition of cell-mediated immunity requires further exploration.

Objective To investigate the protection of simvastatin on monocrotaline (MCT)-induce pulmonary hypertension (PH) and the mechanism thereof. Methods Tirty-two male Sprague-Dawley rats were randomly divided into 4 equal groups: PH group undergoing subcutaneous injection of MCT and then gastric infusion of normal saline (NS) once a day for 21 days, simvastatin control group undergoing subcutaneous injection of NS and then gastric infusion of simvastatin 2μg/g once a day for 21 days, simvastatin intervention group undergoing subcutaneous injection of MTS and then gastric infusion of simvastatin 2 μg/g once a day for 21 days, and control group undergoing subcutaneous injection and gastric infusion of NS. Three weeks later the mean pulmonary arterial pressure (mPAP) was detected by right heart catheter. Then the rats were killed with their lungs taken out. Arterial wall area/vessel area (W/V), and arterial wall thickness/vessel external diameter (T/D) were calculated. Perivascular inflammation was scored with the subjective scale of 0 (no) to 4 (severe). Pulmonary interleukin (IL)-6, tumor necrosis factor α (TNF-α), and monocyte chemotactic protein 1 (MCP-1) were tested by ELISA. Results The mPAP of the simvastatin intervention group was (23 ± 7) mm Hg, significantly lower than that of the PH group [(34 ± 9) mm Hg , P < 0. 05], but not significantly different from that of the normal control group [(20±4) mm Hg, P>0.05]. The W/V and T/D of the simvastatin intervention group were 0.442 ± 0. 061 and 0. 325 ±0. 045 respectively, significantly lower than those of the PH group (0. 560 ±0. 086 and 0. 368 ± 0. 055 respectively, P < 0. 01 and P < 0. 05) . The perivascular inflammation score of the simvastatin intervention group was (2. 19±0. 81), significantly lower than that of the PH group (3.40 ± 0. 65, P < 0. 05), and the IL-6, TNF-α, and MCP-1 levels of the simvastatin intervention group [(264 ± 127), (179 ±91), and (697 ±211) pg/ml respectively] were all significantly lower than those of the PH group [(765±179), (447±86), (4428±757) pg/ml respectively, all P <0.01].Conclusion The protective effects of simvastatin against MCT-induced PH may be associated with the inhibition of the perivascular inflammation and lung IL-6, TNF-α, and MCP-1 levels.

Objective To investigate the effects of reversal of hyperglycemic toxicity on the synthesis and secretion of glucagon by the a cells and the possible mechanisms thereof. Methods Mouse glucagonoma cells of the line TCl-6 were cultured in medium containing 5.5 mmol/L glucose for 10 days (low glucose group, LG), in medium containing 25 mmol/L glucose for 10 days (high glucose group, HG), in medium with 25 mmol/L glucose for 5 days and then in medium with 5.5 nnnol/L glucose for 5 days (high-low glucose group, HL group), or in medium with 5.5 mmol/L glucose for 5 days and then in medium with 25 mmol/L glucose for 5 days (LH group), Radioimmunoassay was used to detect the glucagon concentration in the supernatant. RT-PCR was used to detect the mRNA expression of glucagon in the TCl-6 cells. Western blotting was used to detect the protein expression of the Snare proteins: syntaxin1A and synaptosome-associated protein (SNAP)-25. Results (1) The glucagon level of the HG/LG group was lower than that of the HG group by 29% (P < 0. 05). (2) The glucagon mRNA expression level in the TCl-6 cells of the HG/HL group was significantly lower than that of the HG group by (52. 6 ± 2. 8) % (P < 0.05). (3) The syntaxin1A and SNAP-25 expression levels in the TCl-6 cells of the HG group were significantly higher than those of the LG group by 36% and 69% respectively (both P < 0. 05), and the syntaxin1A and SNAP-25 expression levels in the TCl-6 cells of the HG/LG group were significantly lower than those of the HG group by 49% and 56% respectively (both P < 0. 05). Conclusion After removing the high glucose toxicity the abnormal glucagon secretion by the α-cells can be ameliorated obviously, and the expression levels of syntaxinl A and SNAP-25 proteins that promote the secretion of glucagon are reduced accordingly.

Objective To investigate the effects of ischemic postconditioning (IPest) in protecting hypertrophic myocardium subjected to ischemic-reperfusion (I/R) and to study the role of extracellular regulated protein kinase (ERK1/2) in mediating such protection. Methods Transverse aortic constriction (TAG) operation was performed on 12-week-old C57/BL mice to establish left ventricular hypertrophy models. Sixty-four isolated TAG mouse hearts were mounted onto the Langendorff perfusinn system and randomly divided into 4 equal group: (1) I/R group undergoing stable perfusion for 30 rain, ischemia for 30 min, and re-perfusion for 120 min (an I/R cycle) to cause hypertrophic myocardium I/R injury, (2) IPost group undergoing ischemia for 10s and reperfusion for 10s, totally 3 cycles (60s) before reperfusion for 120 rain, (3) I/R + PD98059 (an ERK1/2 inhibitor) group undergoing perfusion of Krebs-Henseleit(KH) buffer with PD98059 10-5>mol/L for 15 min and perfusion of KH buffer without PD98059 at the beginning of re-perfusion, and (4) IPost + PD98059 group undergoing 3 cycles of IPost and perfusion of KH buffer with PD98059 10-5mol/L for 15 min at the beginning of re-perfusion. Hemodynamic examination was conducted 120 min after re-perfusion to measure the left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), maximal uprising velocity of left ventricle pressure (dp/dtmax), and minimal uprising velocity of left ventricle pressure (dp/dtmin). After the I/R procedure the mvocardium of the left ventricle was isolated to detect the infarction size (IS). Weetem blotting was used to detect the protein expresson of extracellular signal-regulated kinase (ERK) 1/2, phospharylated ERK1/2, Bcl-2, Bax, and mitochondrial and cytosolic eytochrome (Cyt). C. TUNEL was used to detect the apoptotic rate. Results The LVSP and dp/dtmaxlevels of the IPost group were(85 ±4)mm Hg and (3811±230) mm Hg/s, both significantly higher than those of the I/R group [(68 ±5)mm Hg and (2830±230) mm Hg/s respectively, both P < 0.05] .Compared with the I/R group, the protein levels of phosphorylated ERK1/2, Bcl-2, mitochondrial CytC of the IPost group were all significantly higher , the protein levels of Bax and cytosolic CytC, and apoptosis index were significantly lower (all p < 0. 05), and the IS was smaller (P < 0. 05). I/R + PD98059 showed no effects on the above-mentioned parameters. However, the results of the IPost + PD98059 group showed that in the first 15 min of reperfusion addition of PLY)8059 reversed all changes observed in the IPost group and eliminated the IPost protection by increasing IS to a level similar to that in the I/R group. Conclusion IPost has protective effect in hypertrophic myocardium with L/R injury in vitro. ERK1/2 signaling pathway is involved in the protection of IPost by regulating the myocyte apoptosis.

Objective To investigate the anti-tumor effect of eukaryotic expressing plasmid containing human angiostatin Kringle (1 - 3) [hAG (K1 - 3)] combined with soluble tumor necrotic factor-related apoptosis inducing ligand (sTRAIL) genes on human gastric cancer xenografts in nude mice, Methods Recombinant plasmids of pBud-hAG and pBud-hAG-TRAIL were constructed by subcloning technique. Twenty nude BALB/c mice were inoculated with human gastric cancer ceils of the line BGC-823 subcutaneously into the back. One week later after the appearance of implanted tumors the mice were randomly divided into 4 groups with pBud-hAG, pBud-hAG-TRAIL, pBud blank plasmid, and normal saline (NS) injected into the tumors respectively once the other day for 7 times. The size of tumor was observed. 7 days later the mice were killed with their tumors taken out. RT-PCR was used to detect the expression of hAG and sTRAIL The microvessel density (MVD) of tumor was observed and recorded by detecting the protein of CD34 with immunohistiochemitry on the microscopy. Results The MVD of tumor in the pBud-hAG-TRAIL and pBud-hAG groups were (4. 8±0. 9)/HP and (4. 6 ±1.2)/HP respectively, significantly lower than those of the pBud and NS groups [(17.4 ±2.4)/HP and (18.2±2.7)/HP respectively, all P < 0. 05] , but there was no significant difference between the pBud-hAG-TBAIL and pBud-hAG groups. mRNA expression and protein expression of sTRAIL and hAG were positive in the pBud-hAG-TRAIL and pBud-hAG groups. The tumor volumes of tumors of the pBud-hAG-TRAIL and pBud-hAG groups were (1. 325 ± 0. 012)cm3and (1. 862 ± 0. 017)cm3 respectively, both significantly lower than those of the pBud and NS groups [(3. 637 ±0. 032)cm3 and (3.521 ±0.028)cm3 respectively, all P<0.05]. Conclusion Angiostatin inhibits tumor angiogenesis through inhibiting the growth of vascular endothelial cells, and TRAIL induces tumor cell apoptosis, hAG (K1 - 3) combined with TRAIL can inhibit tumor growth more efficiently.

Objective To isolate and purify human islet according to the method established by Ricordi and to evaluate the function and safety of these isolated human islets. Methods Six pancreases were obtained from human corpses. The islets were isolated by liberase digestion and purificated by Ficoll density gradient centrifugation. The numbers, purity and vitality of the islets were analyzed. The various endocrine cell composition and distribution of the islets were checked by immunofluorescence staining. The glucose-induced insulin secretion was detected by chemiluminescence method. The isolated islets were transplanted under the left renal capsules of 10 streptozocin-induced diabetic nude mice. Twenty days later the left kidneys with transplanted islets of 2 mice with normal blood sugar were reseeted, and then blood sugar level was observed. An isolated human islet was suspended in RPMI-1066 culture medium for72 h, then culture of pathogenic micro-organisms, endotoxin and procoagulant activity were detected so as to evaluate the security of the islet products. Results The mean number of the isolated islets was (229 000 ± 31 000) islet equivalents (IEQs)/pancreas or (4970 ± 1620) IEQs/g pancreatic tissue, the mean purity was (59. 0 ± 8.9) %, and the mean vitality was (89 ± 3)% for the purified islets. Immunofluorescence staining showed that there were 4 types of endocrine cells normally distributed in the islets. The mean insulin stimulation index was 8. 1 ±4. 0(3. 8 - 10. 2). The glycemia found in the diabetic nude mice decreased to normal levels from the third day after islet transplantation and maintained normal for over 30 days. The parameters of security in these islet products were under the standard scope. Conclusion Human islets obtained according to Ricordi's method reach the standard for clinical islet transplantation in number, purity, vitality, function, and security.

Objective To investigate the curative effects of inhaling signal transducer and activator of transcription 1 (STAT1) antisense oligonucleotide (ASON) on alveolitis and pulmonary fibrosis and the best administration time. Methods Twenty-five adult female Wistar rats were randomly divided into 5 equal groups: BLM group, undergoing intra-tracheal perfusion of BLM so as to establish animal models of alveolitis and pulmonary fibrosis and then inhaling aerosolized normal saline (NS); NS group undergoing intra-tracheal perfusion of NS and then inhaling aerosolized NS; ASON 0 d group, undergoing intra-tracheal perfusion of BLM and then inhaling aerosolized STAT1 ASON 3 ml immediately; ASON 7 d group, undergoing intra-tracheal perfusion of BLM and then inhaling STAT1 ASON 3 ml 7 days later; and ASON 14 d group undergoing intra-tracheal perfusion of BLM and then inhaling aerosolized STAT1 ASON 3 ml 14 days later. Aerosolized inhalation was repeated once every other day for 4 times. Twenty-eight days after intra-tracheal perfusion the rats were sacrificed with their lungs taken out to undergo pathological examination. NS was infused into the right lungs to get bronchoalveolar lavage fluid (BALF). ELISA was used to examine the concentrations of transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) in the BALF. Results The pathology result of the lung tissues showed that compared with the BLM and ASON 14 d groups, the alveolitis and pulmonary fibrosis of the ASON 0 d group were obviously milder. The scores of alveolitis and pulmonary fibrosis of the ASON 0 d group were (1.80 ± 0. 84) and (2. 60 ± 0. 55) respectively, both significantly lower than those of the BLM group [(2.40 ± 0. 55) and (4. 40 ± 0. 55) respectively] and those of the ASON 7 d group [(2. 20 ±0. 45) and (3.00 ±0. 71) respectively] (all P < 0. 05). The scores of pulmonary fibrosis of the ASON 7 d group was significantly lower than those of the BLM and ASON 14 d groups (both P < 0. 05). The concentrations of TGF-β and TNF-α in BALF of the ASON 0 d group were (48. 11 ±3.46) pg/ml and (1.93 ± 0. 14) ng/ml respectively, both significantly lower than those of the BLM group [(57. 67 ± 2. 46) pg/ml and (2. 45 ± 0. 25) ng/ml respectively, both P < 0. 05]. The concentration of TGF-β in BALF of the ASON 0 d group was significantly lower than those of the ASON 7 d and ASON 14 d groups [(51.42 ±3. 57) pg/ml and (55.8 3 ± 1.79) pg/ml respectively, both P <0. 05]. The concentration of TGF-β in BALF of the ASON7 d group was significantly lower than those of the BLM and ASON 14 d groups (both P < 0. 05). Conclusions STATI ASON administered in the early stage helps depress the pulmonary fibrosis procedure, and the earlier the drug is administrated the better effect would be obtained. Aerosolized STATI ASON can be used as a therapeutic method for pulmonary fibrosis.

Objective To prepare rabbit polyclonal antibodies against intraceUular peptides of human platelet glycoprotein GPIbα. Methods Two peptides corresponding to human platelet GPIbα C-terminus were synthesized and purified by high-performance liquid chromatography (HPLC). The peptides were cross-linked with keyhole limpet hemocyanin (KLH). Two New Zealand white rabbits were immunized with conjugated peptides for 3 times. The polyclonal antibodies were purified by Ammonium Sulfate Precipitation and identified by dot blotting and ELISA. GPIbot intracellular poptides phospharylation was tested with these polyclonal antibodies by ELISA. Results The titers of the two polyclonal antibodies against the GPIbα C-terminus poptides were 1 : 32 000 and 1 : 64 000 respectively and both of these antibodies reacted with purified GPlbot. Conclusions Two rabbit polyclonal antibodies against C-terminal peptides of human platelet GPIbα have been prepared successfully, providing a way for the preparation of these kinds of antibody. Both phosphorylation and dephosphorylation states exist in the intracellular poptide of human platelets.

A 43-year-old female was admitted to the hospital on 24 September 2003 due to epigastric pain accompanied by fever for 1 month and aggravated for 2 days. One month ago, she developed epigastric pain without obvious trigger, paroxysmal, radiating to the back, accompanied by chills, fever, body temperature up to 40℃, accompanied by vomiting, the vomit was stomach content, and the symptoms did not improve after intravenous infusion of ofloxacin and metronidazole, and her body weight lost 10 kg.

Patient Xu × ×, male, 62 years old. Two years ago, he felt that his abdomen was gradually enlarged without obvious trigger, but recently, due to the gradual aggravation of abdominal bulge, CT examination showed: huge cystic-solid mixed density foci in the abdominal cavity, and chaotic vascular shadows were found on the upper edge of the tumor on enhanced scan, the size was 29 cm ×22 cm, the boundary with the right lobe of the liver was unclear, and the surrounding organs were obviously compressed and displaced. He went to our hospital for treatment, and was admitted to the hospital with a huge abdominal mass in the outpatient department. He denied his history of hepatitis and had no tobacco and alcohol habit.

The birth of coronary angiography ushered in a new era of percutaneous coronary intervention in the treatment of coronary heart disease, and the appearance of drug-coated stents further consolidated the position of coronary intervention. However, coronary angiography is not perfect, its biggest defect is that it cannot show the blood vessel wall, and the most basic pathological change of atherosclerosis is precisely the change of blood vessel wall. Intravascular ultrasound can see the blood vessel wall but must insert a catheter into the coronary artery, which greatly hinders its wide application.