Objective　To investigate the relationship between the tumor angiogenesis and the clinicopathological variables of hepatocellular carcinoma (HCC) and determine whether tumor angiogenesis can be used as an indicator for predicting invasion, metastasis and prognosis of HCC. Methods　Using the immunohistochemical S-P method, CD34 expression was detected in liver tissue from 20 normal individuals, 20 patients with liver cirrhosis and 85 patients with HCC and juxtacancerous tissues and the microvascular density (MVD) determined in those from 85 patients with HCC. Results　CD34 was mainly located in the cell cytoplasm of endothelial cells. The CD34 staining was confined to vessels in the portal triad in the above-mentioned various tissues. In HCC tissues, however, sinusoid-like vessels reacted intensively with anti-CD34 and the MVD was 156.5±62.4 (per 0.74 mm2). MVD was correlated to tumor size, number, differentiation, emboli in the portal vein and capsule (P<0.05 or 0.01) but not to HBsAg and AFP (P>0.05). It was also correlated with prognosis of HCC. The mean tumor-free survival of MVD ≥156 was 14 months while that of MVD <156 was 53 months (P<0.01). Conclusion　The expression of CD34 in sinusoid-like vessels in HCC tissues represents the neovascularization. Angiogenesis in HCC is associated with the progression of HCC. MVD can be used as an indicator to judge the invasion, metastasis and prognosis of HCC.

Objective To investigate the alteration of lymphocyte P-gp expression level and function in liver transplant recipient and its effect on immunosuppressive therapy.Methods Using flow cytometry,the level of lymphocyte P-gp and analyzed the P-gp function in 53 cases of liver transplant recipieints was observed.Results Among 53 cases,the preoperative mean level of lymphocyte P-gp was(10.34?4.0)%,P-gp level began to increase in the 1st month postoperatively(17.8?8.0)%,peaked in the 3rd month(27.5?13.3)%,and then was stable at this level thereafter.There were no significantly difference between different sex of age group.But the P-gp level in CD+4T cells was significantly higher than that in CD+8T cells and B cells,RH123test showed that high P-gp expression of lymphocyte enhanced its transport activity of medical agents,and decreased the intracellular accumulation of drug more significantly than that of low P-gp expression(P

Objective To investigate the effects of probucol and losartan on cell proliferation after balloon angioplasty in hypercholesterolaemic rabbits.Methods Forty male New Zealand rabbits were randomly divided into high cholesterol diet group,probucol group,losartan group and combined drugs group.The iliac arteries of the rabbits were balloon-injured and the insulin-like growth factor-I receptor(IGF-IR),vascular endothelial growth factor(VEGF),and proliferating cell nuclear antigen(PCNA) were detected by using immunohistochemical method.Results Compared with high cholesterol diet group,the lumen areas were enlarged and the intimal areas were decreased in the probucol group,losartan group and combined drugs group(all P

Objective To choose one protocol that can quickly ,safely and effectively provide amount enough of bone marrow derived mesenchymal stem cells(MSCs) for use of clinical or experimental test through comparison of their growth characteristics and growth factors levels in culture solution .Methods Cells extracted from bone marrow of C57BL/C mice respectively underwent two different isolation protocols :whole bone marrow adherent culture(WBMAC) or gradient density separation(GDS);characteris‐tic surface antigens of MSCs were identified by flow cytometry on cells isolated in different ways ;the distinct growth curve of pri‐mary stem cells cultured in vitro described their different proliferation rate;levels of vascular endothelial growth factor(VEGF) and stromal cell‐derived factor‐1α(SDF‐1α) in culture medium were detected by ELISA .Results Primary MSCs obtained by WBMAC proliferated at higher speed and exhibited shorter growth cycle than those separated by GDS ;on MSCs from both groups ,surface antigens CD29 were detected positively ,and antigens including CD31 ,CD34 and CD45 were assayed negatively ;concentration of VEGF and SDF‐1αin both two nutrient solution primarily keep at low levels ,comparatively ,level of VEGF and SDF‐1αin the di‐shes which contain MSCs by WBMAC was higher than the one in the dishes which contain MSCs by GDS .Conclusion MSCs ex‐tracted by WBMAC shows unimpaired cell function ,can build automatically more suitable microenviroment for their growth;this classic method was qualified for clinical and experimental use in a safe ,rapid ,effective way .

Objective To investigate the influence and the clinical significance of interleukin-2 (IL-2) on T lymphocyte subgroup after chemotherapy in acute myeloid leukemia patients. Methods Fiftyfour acute myeloid leukemia patients were divided into treatment group (chemotherapy combined with IL-2,28 cases) and control group (pure chemotherapy,26 cases) by radom digits table. In treatment group, IL-2400 000 U was dripped after chemotherapy for 14 days. Then the T lymphocyte subgroup change before and after treatment was examined. Results After 14 days' treatment, the levels of CD3 (0.6026±0.2275 ),CD4(0.4972±0.1224),CD56(0.3016±0.1053 ) in treatment group were significantly higher than those before treatment(0.3926±0.2010,0.2264±0.1190,0.1729±0.1226) and in control group (0.4352±0.1930,0.2738±0.1362,0.1937±0.1268)(P< 0.05). Conclusion IL-2 treatment can obviously raise the levels of T lymphocyte subgroup and can raise the immunologic function.

Objective To investigate the effect of mild hypothermia on the expression of hypoxia-inducible factor-1α (HIF-1α) and glucose transporter-1 (GLUT-1) in a rat model of chronic cerebral ischemia-reperfusion.Methods Thirty-six female SD rats weighing 170-210 g were randomly divided into 3 groups (n = 12 each):sham operation group (group S), normal body temperature group (group NT ) and mild hypothermia group (group MH). Arterio-venous fistula was established by end-to-end anastomosis between the right common carotid artery and right external jugular vein for 6 weeks followed by reperfusion. In group MH, mild hypothermia was induced at the initiation of reperfusion and the rectal temperature was reduced to 31.5-32.5 ℃. In group S and NT, the rectal temperature was maintained at 37-38 ℃. Six rats in each group were sacrificed at 3 and 48 h of reperfusion. The brains were immediately removed for determination of the expression of HIF-1α, GLUT-1, HIF-1α mRNA and GLUT-1 mRNA and microscopic examination. Results Compared with group S, the expression of HIF-1α and HIF-1α mRNA at 3 and 48 h of reperfusion and GLUT-1 mRNA at 3 h of reperfusion was up-regulated, while the expression of GLUT-1 and GLUT-1 mRNA at 48 h of reperfusion was down-regulated in group NT (P ＜ 0.05).Compared with group NT, the expression of HIF-1α and HIF-1α mRNA at 48 h of reperfusion and HIF-1α mRNA and GLUT-1 mRNA at 3 h of reperfusion was down-regulated, while the expression of GLUT-1 and GLUT-1 mRNA at 48 h of reperfusion was up-regulated in group MH (P ＜ 0.05).Microscopic examination showed that the injury to the ultrastructure of blood-brain barrier was significantly attenuated in group MH compared with group NT. Conclusion Mild hypothermia can attenuate chronic ischemia-reperfusion injury by down-regulating the expression of HIF-1α and up-regulating the expression of GLUT-1.

BACKGROUND:The preliminary findings confirm that bone marrow mesenchymal stem celltransplantation is safe and effective in the treatment of acute myocardial infarction, but its exact mechanism is unclear. There are few studies addressing the survival status of transplanted stem cells and its acting timing.
OBJECTIVE:To study the survival of rat bone marrow mesenchymal stem cells transplanted into the infracted myocardium. METHODS:Bone marrow mesenchymal stem cells were cultured using density gradient centrifugation. Eighty rat models of myocardial infarction were prepared. Bone marrow mesenchymal stem cells were injected via a microsyringe at four sites around the infarcted region at 14 days after modeling. Then, 70 rats with living cells were selected for detecting the survival of bone marrow mesenchymal stem cells at days 3, 5, 7, 10, 20, 28 after transplantation. RESULTS AND CONCLUSION:Under ×400 visual field, the number of Brdu-positive bone marrow mesenchymal stem cells was (36±12) at 3 days posttranplantation, (33±13) at 5 days, (28±9) at 7 days, (15±5) at 10 days, (5±3) at 14 days, 0 at 20 days, and 0 at 28 days, showing a overal downward trend after transplantation. The number of bone marrow mesenchymal stem cells was negatively correlated with transplant days (P<0.01, r=-0.47). The number of cells decreased most significantly within 1 week after transplantation, and then decreased to 0 at 20 days. These findings indicate that transplanted bone marrow mesenchymal stem cells in the myocardium cannot survive for a long term and also cannot be transformed into myocardial tissue.

BACKGROUND: Bushen Huoxue Tongluo Capsule, a well-known traditional prescription, exhibits remarkable treatment outcomes in knee osteoarthritis, but the underlying mechanism is not fully understood. OBJECTIVE: To explore the regulatory effect of Bushen Huoxue Tongluo Capsule on miR-27a of synovial cells in Sprague-Dawley rats with knee osteoarthritis. METHODS: The model of knee osteoarthritis was established in 120 Sprague-Dawley rats using Hulth method, and then these rat models were randomized into four groups, followed by given75 mg/kg normal saline (blank control group), 34 (low-dose group), 75 (medium-dose group) and 140 (high-dose group) mg/kg Bushen Huoxue Tongluo Capsule aqueous solvent via gavage according to body mass, respectively, twice daily, for consecutive 2 courses (15 days as a course) with 2 days in between. The synovium and cartilages were removed after each course, and the expression of miR-27a in the knee joint synovium and chondrocytes was determined by real-time quantitative PCR. RESULTS AND CONCLUSION: Gross observation found that the degeneration of synovium and cartilage in the Bushen Huoxue Tongluo Capsule groups was milder than that in the blank control group. Real-time PCR revealed that there was an increasing trend in the expression of miRNA-27a in the synovium and cartilage in each group, especially in the high-dose group, and there was a significant difference between groups (P < 0.05). Our findings show that Bushen Huoxue Tongluo Capsule can suppress the reduced expression of miRNA-27a in the synovium and chondrocytes in rats with knee osteoarthritis, further alleviating knee degeneration and protecting the knee from damage.

BACKGROUND: Zero-P anterior cervical fixation system is an important means for treatment of cervical myelopathy in recent years. Zero-P fixation system can achieve effective decompression of spinal cord and nerve, and restore cervical curvature, with good stability.OBJECTIVE: To investigate the biocompatibility of Zero-P for multilevel cervical myelopathy.METHODS: Totally 62 patients with multilevel cervical disease who were treated by anterior cervical discectomy and fusion with Zero-P were selected, including double segments in 47 cases, three segments in 13 cases and four segments in 2 cases. Clinical efficacy was evaluated by pain visual analogue scale score, Japanese Orthopedic Association score, neck disability index score at postoperative 1, 6, 12, 24 months. Simultaneously, incidence of postoperative complications, cervical intervertebral space height and Cobb angle changes were observed.RESULTS AND CONCLUSION: (1) 54 patients were followed 2 years after treatment by clinic or telephone. Mean operative time was (102.00±32.41) minutes; average blood loss was (62.45±18.36) mL. (2) Two patients affected mild throat discomfort at one day after operation, and the symptoms went to lift six days after. The remaining patients did not experience any other complications. (3) Visual analogue scale score and neck disability index score at 1, 6, 12 and 24 months after operation were all less than preoperatively (P < 0.05); Japanese Orthopedic Association scores were higher than preoperatively (P < 0.05); the Cobb angle was improved significantly and greater than before treatment (P < 0.05).Postoperative cervical intervertebral space height was significantly higher than preoperatively (P < 0.05). Japanese Orthopedic Association results showed that among 54 patients, excellent was found in 34 cases, good in 14 cases,average in 6 cases and poor in 0 cases; the excellent and good rate was 89%. (4) Zero-P has a good clinical efficacy for multilevel cervical myelopathy, relieves symptoms significantly and restores cervical curvature and height, and has a good biocompatibility.

Objective To investigate the effects of uric acid (UA) and UA under oxidative stress on cultured human umbillical vein endothelial cells (HUVEC).Methocds HUVECs were incubated with different concentration UA (0,4,8,16 mg/dL),H2O2 (500 mmol/l) and UA+H2O2 (500 mmol/l) for 24,48 and 72 hours.Then we observed the morphology of HUVECs and evaluated the proliferation of HUVECs by MTT assay.NO and ET-1 in supernatant medium was detected by ELISA.Results For the viability of HUVECs,there was no statistically significant difference between 4 mg/dL UA group and control group after incubation for 24,48 and 72 hours (P＞0.05) and between UA groups(8 mg/dL and 12 mg/dL) and control group after incubation for 24,48 and 72 hours (P＞0.05).After incubation with 12 mg/dL of UA for 48 hours or 8 mg/dL of UA for 72 hours,the viability of HUVECs decreased significantly (P ＜0.05) The viability of HUVECs in H2O2 group decreased significantly (P＜0.05).The viability of HUVECs in UA+H2O2 groups after incubation for 24 h was significantly better than H2O2 group.There was no signifiant difference in the cell viability between (8 mg/dL or 12 mg/dL) UA+H2O2 group and H2O2 group.Compared with the control group,the NO levels were decreased and the ET-1 levels were increased in the supernatants of HUVECs in 12 mg/dL UA group for 72 hours (P＜0.05).Compared with H2O2 group,the NO levels were increased and the ET-1 levels were decreased in the supematants of HUVECs in (8 mg/dL or 12 mg/dL) UA +H2O2 groups for 24 hours (P＜0.05),while for (12 mg/dL) UA +H2O2 group for 72 hours,the results were just the contary.Conclusion The effects of UA on HUVECs are related with both concentration and action rime.Acutely increased UA may protect HUVECs form injury,while long action of UA may injure HUVECs,especially under oxidative stress.