Objective: To detect the expressions of CD 44V6 and β-catenin in osteosarcoma and to analyze the association of their expressions with migration and invasion of osteosarcoma cells. Methods: Expressions of CD 44V6 and β-catenin were examined by immunohistochemical SP method in 35 specimens of human osteosarcoma tissues, 15 specimens with osteochondroma, and 15 cases of normal bone tissues. Results: The positive expressions of CD 44V6 and β-catenin were significantly higher in osteosarcoma than osteochondroma and normal control tissues (P < 0.01). Their expressions significantly correlated with clinical staging, tumor metastasis, and 2-year death (P < 0.05); but had no correlation with the age and gender of patients, tumor size, histological classification, and with or without preoperative chemotherapy (P > 0.05). In osteosarcoma, expression of CD 44V6 was positively associated with the expression of β-catenin (P < 0.01). Conclusion: The positive expressions of CD 44V6 and β-catenin closely correlate with the occurrence, development, and metastasis of osteosarcoma. They may play synergistic role in the invasion and metastasis of osteosarcoma. Combined detection of CD 44V6 and β-catenin may be used as an indicater for diagnosis and prognosis of metastasis of osteosarcoma.

OBJECTIVETo search for an optimal protocol and freezing conditions for the cryopreservation of microamount round spermatids of the mouse.

METHODSWe compared the survival rates of frozen-thawed microamount round spermatids of the mouse achieved by vitrification or standard slow freezing with different concentrations of glycerol (5, 7, or 9%) and different lengths of equilibrium time (0, 15, 30, 45, or 60 min).

RESULTSUnder the conditions of 7% glycerol and 30 min equilibrium, both vitrification and standard slow freezing achieved high survival rates of spermatids, and the former obtained an even higher rate than the latter ([72.9 ± 15.4]% vs [58.2 ± 17.7]%, P < 0.05).

CONCLUSIONA high rate of frozen-thawed microamount round spermatids of the mouse can be achieved by vitrification under the conditions of 7% glycerol and 30 min equilibrium.

Animals ; Cell Survival ; Cryopreservation ; methods ; Cryoprotective Agents ; administration & dosage ; pharmacology ; Glycerol ; administration & dosage ; pharmacology ; Male ; Mice ; Spermatids ; Time Factors ; Vitrification ; drug effects

In this paper, based on the scientific research of TCM for many years and from the front-line point of view, the author explored the mainly common problems which had been existed in the TCM research, such as the establishment of the quality standard, the research of innovative drugs, the research ideas differentia between TCM and the natural drugs, the improvement of the research management model and other issues.

In this paper, the author proposed the development direction and study task for processing subject. Its development direction were study of scientific research method and establishment of technological platform for processing-patterns for the scientific research and production combination (enterprise as the mainstay). And its main task will be located at the inheritance of the traditional Chinese medicine pharmaceutical technology, the protection of processing culture as the non-material heritage, and the development of processing as a modern subject.

Objective To evaluate the effect of alIoHCT on recurrence and metastasis of HCC after hepatic radical resection. Methods Umbilical cord blood were collected after labor. The efficacy of separa-tion by 6% hydroxyethyl starch (6% HES) and NH4CL lysing solution were examined. Twenty-two SCID mice were randomized into the scheduled transplantation group (n=8), the single transplantation group (n =8) and the normal saline group (n=6). Human nucleated cells (NC)at a amount of 5×107 were transfused through the tail vein into the 2 groups of transplantation. All the mice in the 3 groups received cyclophospha-mide (CTX) as conditioning regimen before tansplantation and Methylprednisolone (MP) for 1 week continu-ously after trasplantation. Hematopoietic and immune recovery, graft versus host disease (GVHD), engraft-ment and survival rate were observed after transplantation. Six weeks after alloHST, the orthotopic tumor model in SCID mice was established by implanting histologically intact tissue under the embrane of liver. Ten days later, the mice received radical resection of lobe bearing tumor. The condition of recurrence and metas-tasis was observed 4 weeks after operation. Results A murine model of umbilical cord blood transplantation using CTX and MP as conditioning regimen could be successfully established in SCID mice. The percentage CD34+ cells of peripheral blood NC in scheduled transplantation group and single transplantation group was 1.66%±0.47% and 0.68% + 0.56%, respectively. There was significant statistical significance (P<0.01). The intrahepatic recurrence rate after operation was 100% in all the 3 groups. However, the recur-rent tumor volume was (367.18±31.86) mm3 , (648.26±155.22) mm3, (811.38±127.36) mm3, re-spectively in the 3 groups. There was marked difference among the 3 groups (P<0.01). The inhibitory rate of group A and B was 54.7% and 20.1%. The incidence of lung metastasis was 14.3 (1/7), 6.7% (4/6) and 100% (5/5), respectively in the 3 groups and there was remarkable difference (P<0.01). The inci-dence of celiac lymph node metastasis was 14.3 (1/7), 33.4% (2/6) and 40% (2/5), respectively in the 3groups and there was no significant difference (P=0.58). Conclusion AlloHSCT is a useful method for de-creasing metastasis and recurrence of liver cancer after radical resection in early stage.

A method was developed for determining residual dexamethasone and betamethasone in swine liver by HPLC-MS/MS with isotope dilution. The samples were digested by β-glucuronidase/aryl sulfatase, and extracted) with acetonitrile. Further cleanup was performed on C_(18)) cartridge and liquid-liquid extraction with sodium) carbonate solution. Then the supernatant was dried under nitrogen and residues were dissolved in mobile) phase. The samples were analyzed by HPLC-MS/MS on a Hypercarb C_(18)) column with a mixture of acetonitrile-water-formic acid as the mobile phase. The samples were quantified with the internal standard calibration curve method using isotope dilution. The limit of detection for dexamethasone and betamethasone in swine liver was 0.12 μg/kg and 0.14 μg/kg, respectively. The limits of quantification were 0.42 μg/kg and0.47 μg/kg), respectively. The average recoveries were 97.3%-111.0%, the intra-assay relative standard deviations were 1.85%-5.65% and the inter-assay relative standard deviations were 2.8%-8.0% at spiked levels of 0.75-2.00 μg/kg. There was a good linear correlation (the correlation coefficient is above 0.9997) between the ratio of peak areas of quantitative ion-pair to internal standard and concentration of analyte) in the range of 10-500 μg/L.

Objective To investigate if mitochondrial KATP channels are involved in the cardioprotective effects of astragalus membranaceus in immature rabbit hearts. Methods Twenty-four New Zealand white rabbits of both sexes aged 14-21 days weighing 300-350 g were anesthetized and heparinized. The hearts were rapidly removed after thoracotomy and mounted on a Langendorff apparatus via ascending aorta and perfused with oxygenated (95% O2-5% CO2) Krebs-Hensleit buffer (KHB) solution at 60 cm H2O (perfusion pressure) and 38 ℃ . A fluid-filled latex balloon was inserted into left ventricle via left atrium for measurement of left ventricular developed pressure ( LVDP) , Global myocardial ischemia was induced by suspension of perfusion for 30 min followed by 45 min reperfusion. The animals were randomly divided into 3 groups with 8 animals in each group : group A control;group B astragalus and group C astragalus +- 5-HD (a selective inhibitor of the mitochondrial KATP channel). In group B the hearts were perfused with astragalus 40 g? L-1 for 15 min before ischemia. In group C the hearts were perfused with 5-HD 100?mol?L-1 for 5 min followed by 10 min astragalus perfusion before ischemia. In control group the hearts were perfused with only KHB before ischemia. Coronary flow (CF), HR, LVDP and ? dp/dtmax, were measured at 5, 10, 15, 30 and 45 min of reperfusion and recorded. Coronary effluent was collected at 10 min of reperfusion for determination of CK-MB, LDH and CK levels . At the end of 45 min reperfusion the isolated heart was removed for determination of myocardial ATP and iNOS levels and electron microscopic examination.Results In astragalus group (B) LVDP ? dp/dtmax and CF recovered significantly better and myocardial ATP content was significantly higher as compared with group A and C (P

Objective: To observe the changes of circulating endothelial cells (CEC) and endothelin(ET) and the effect of aprotinin on them during open heart surgery. Method:Twenty patients undergoing valve replacement were divided into two groups,control group(n=10) in which and protinin group (n=10), 4000000 unit aprotinin were given during cardiopulmonary bypass (CPB), Blood samples were collected before operation,pre-CPB,15,30 and 60 minute following CPB and 1st, 3rd, 5th day after operation. Result: As compared to pre-CPB in control group,CEC and ET levels increased during CPB and returned the normal after operation. In aprotinin group,the tendency of change was same,but CEC and ET levels were higher in control group than in aprotinin group during CPB. Conclusion:There are endothelial ceils damaging and ET significant increasing during CPB,while aprotinin can ameliorate this situation.

Objective To evaluate the efficacy and safety of simulect (basiliximab) after orthotopic liver transplantation (OLT). Methods Forty adult recipients with benign end-stage liver disease between November 2003 to November 2004 were assigned randomly in a 1∶1 ratio to receive either two doses of simulect or matching placebo. The patients in the two groups received baseline triple immunosuppression with the calcineurin inhibitor (CsA or FK506), MMF and steroids. A total of 40 mg simulect was given in two doses of 20 mg each on the day 0 before inferior vena was opened and the day 4 after transplantation respectively. Acute rejection, infection and serum ALT, AST, TBIL, DBIL and ALP in both groups were observed in the first 30 days after OLT. Results In Simulect group had less frequent incidence of acute refection during the first 30 days after OLT. In Simulect group and matching placebo, incidence of acute refection was 10 % (2/20) abd 45 % (9/20) respectively (P= 0.034), and that of infection was 40 % (8/20) and 45 % (9/20) respectively (P= 0.749). Bilirub and aminopherase in both groups were declined gradually and ALP increased. There were significant difference in ALT and TBIL between two groups. Conclusion The application of simulect in combination with CsA/FK506, MMF and prednisone is safe and well tolerated, and can effectively reduce the incidence of acute refection, and does not lead to increased opportunistic infections.

Leiomyomas are benign soft tissue swellings of smooth muscle origin, most commonly found in the uterus. Extra uterine leiomyomas presenting as an abdominal mass is often a diagnostic challenge as such occurrence is rare. We present a rare case of primary abdominal wall leiomyoma, and highlight the importance of laparoscopic approach in the diagnosis and treatment of such tumour.
Leiomyoma