Objective To investigate the fluctuated pattern of endogenous bactericidal/permeability-increasing protein(BPI)levels,and to elucidate its significance in endotoxin response after major abdominal surgery.Methods Eleven patients undergoing major abdominal surgery were involved in present study.The levels of plasma BPI,lipopolysaccharide-binding protein(LBP)and interleukin-6(IL-6)were measured preoperatively and on postoperative days 1,3,5,7,and 14 by ELISA.Plasma endotoxin concentration was determined with the chromogenic limulus amebocyte lysate(LAL),which was modified by perchloric acid(PCA)pretreatment of samples.The polymorphonuclear neutrophil count was also done at each time point.Results In comparison with normal controls,plasma BPI/LBP ratios [(1.058?0.074)?10-3,P=0.004] were significantly increased on postoperative day 1,maintained at same level on day 3,and declined on day 5.No plasma IL-6 was detected from the blood of all the 11 patients prior to operation,but plasma IL-6 level rose rapidly on day 1 after operation(27.440?12.144pg/ml,P=0.04),maintaining a high level(11.530?5.312pg/ml)on postoperative day 3,and disappeared on day 5.Conclusions Plasma BPI/LBP ratios were markedly elevated during 3 days after a major abdominal surgery,and endogenous elevated BPI may play an important role in prevention from the development of endotoxemia in surgical patients.

ObjectiveTo investigate the relationship between sepsis induced high mobility group 1 (HMG 1) gene expression and endotoxemia in rats. MethodsSepsis was induced by cecal ligation puncture (CLP). One hundred Wistar model rats were randomly divided into four groups: normal control group ( n =10), sham operation (SO) group ( n =10), CLP group (further subdivided into 2?h, 6?h, 12?h, 24?h, 48?h and 72?h post CLP, n =10 in each subgroup), and bactericidal/permeability increasing protein(rBPI 21 ) treatment group (further subdivided into 12?h, 24?h post CLP, n =10 in each subgroup). Tissue samples from the liver, lungs and kidneys were harvested for the measurement of endotoxin concentrations and HMG 1 mRNA expression by RT PCR. Results Compared to normal control and SO groups the HMG 1 mRNA levels significantly increased in tissues during 6～72?h post CLP ( P

Objective To establish a satisfactory lung metastasis model of human choriocarcinoma using severe combined immunedeficient(SCID)mice and explore the appropriate cell concentration for the model.Methods Forty SCID mice aged between 5-6 weeks were randomly difided into four groups.1×107 cells/ml ×0.1 ml.5×106 cells/ml×0.2 ml and 1×106 cells/ml×0.1 ml of human choriocarcinoma cells JEG-3 were respectively injected in SCID mice of experimental groups by lateral tail vein,the remain group was assigned to the control group.The status and weisht of mice were observed every three days.When these mice were being dying.the size and the number of the lesions of lung metastasis in every mouse were inspected with Micro CT.After Micro CT inspection,the SCID mice were executed dissected to note whether there were tumors on all organ surfaces witll naked eyes.then made pathological sections from the metastaticfoci of fresh lung tissues,and cultured primarily cells and purified cells and passaged cells isolated from the same metastastic foci.The pathological sections were observed under the microscope.The special antigen human chorionic gonadotropin-beta subunit(β-hCG)of the choriocareinoma cells was immunohistochemically detected in the pathological sections and the cells out of cultured primarily cells.The chromosomes of the cells out of cultured primarily cells were analysed.Results Of the group inoeutated 1×107 cells/ml×0.1 ml.all mice died when inoculating.In the group of 5×106 cells/ml×0.2 ml,when inoculating, 3 mice died; the remain 7 mice were being dying on ( 18. 0 ±2. 0) days after injection. 5 of them, there were 1 - 3 lesions of lung metastasis after Micro CT inspection in each mice, and the diameter of the tumors lesions reached 1.5 - 3.5 ram, which was choriocarcinoma confirmed by pathological sections.The special antigen β-hCG was detected by immunohistoehemical method in the pathological sections of pulmonary tissue with tumor and in the cells, which were purified and passaged from being cultured primarily cells isolated from metastastic foci of fresh lung tissues from the SCID mice. The chromosome numbers of these cells out of cultured primarily ceils were variety from 19 to 128, and medal numbers were variety from 70 to 79. Conclusions We successfully established the lung metastatic model of human choriocarcinoma in SCID mice by injecting JEG-3 cells into lateral tail vein, of which 5 × 106 cells/ml × 0. 2 ml is the suitable concentration and volume for the model.

Objective To establish the high-performance liquid chromatography(HPLC)method for measuring concentrations of methotrexate(MTX)in rat lung and some other tissues through internal iliac artery infusion.Methods Fifly female Sprague-Dawley rats were included in this study.The rats were randomly assigned to two groups.Methotrexate was injected to group one through internal iliac artery,and was injected to group two through femoral vein.Blood and tissues were collected in each group at 15,30,60.90 and 120 minutes for detection of the drug concentrations with HPLC.Results The area under the concentration time curve(AUC)in rat lung,ovary and uterus in the artery group were separately(3.77±0.28),(4.40±0.40),(9.97±0.89)μg·h-1·g-1,which were significantly different from those of the vein group[(2.31±0.25),(3.91±0.19),(7.65±1.54)μg·h-1·g-1;P<0.05].The AUC in the rat plasma,heart,kidney,liver and spleen in the artery group were separately(6.13±0.53),(1.90 ±0.11),(5.32±0.89),(14.16±1.96),(0.76±0.20)μg·h-1·g-1.There were no significant differences from the vein group[(5.79±0.71),(1.64±0.29),(5.15±1.69),(14.29 ±3.47),(0.76±0.13)μg·h-1·g-1;P>0.05].Conclusions Through internal iliac artery infusion,there are higher drug concentrations in lung.uterus and ovarian compared to venous injection.The internal-arterial chemotherapy may be used to treat pulmonary metastasis of gynecological tumor.

As a vital component in an overall laboratory animal care and use program , development of a disaster plan plays a critical role for every research institution .Currently, most of domestic institutions would draw up an“emergency operation plan , EOP”, but ignoring a practicable “business continuity plan , BCP” in establishing a disaster plan.In this article, we will discusse about the definition of disaster , how to set up an EOP, and how to establish a thorough BCP , in order to show an integrated and professional disaster plan in laboratory animal care and use program .

Forty patients who underwent sub-total gastrectomy were divided into four groups, one of them for comparison, the other three groups received 10%, 30% and 100% branched-chain amino acid (BCAA) respectively. The results showed that in BCAA-10 and BCAA-3H group, negative balance was partially corrected, but in BCAA-10 group nitrogen remained an obvious negative balance on the 3rd postoperative day. There were more than ten of plasma AA lower than preoperative day in these two groups. In BCAA-30 group, after surgery, negative nitrogen balance was significantly corrected, and changes of plasma AA had no statistic significance compared with that before operation.Creatinine of urine had no significant difference between each group.In BCAA-30 group, transferrin (Tf) was increased compared with that before operation. It is suggested that the protein synthesis is corrected but the catabolism is not restrained.BCAA may play a regulatory role in muscle protein metabolism, BCAA can serve as fuel substrate to promote synthesis and reduce breakdown of muscle protein to correct negative nitrogen balance.Application of large quantities of BCAA should be based on isotonic AA solution.

Objective To assess clinical features and treatment of angiosarcoma.Methods A retrospective study was carried out among seven patients hospitalized for angiosarcoma in the Department of Plastic Surgery,Zhongshan Hospital of Fudan University from January 2004 to March 2012.Clinical behaviors,therapeutic strategies and outcomes of angiosarcoma were analyzed in these patients.Results Of the seven patients,four were male,and three were female,with a median age of 67 years (range,37-77 years).Clinical presentations were various,including irregular ulcerous nodular masses in four patients,localized ulcer in one patient,and irregular erythematous nodules in two patients.All the patients were treated surgically with or without postoperative adjuvant therapy.Pathological examination revealed various degrees of differentiation and atypia of tumor cells,as well as obvious proliferation of endothelial cells,and all of these patients were diagnosed as angiosarcoma.Immunohistochemical analysis revealed a positive staining for CD31 and CD34.After surgical treatment,angiosarcoma recurred in four patients with the median time to the first recurrence being 3.5 months (range,1-6 months),cerebral metastasis occurred in one patient 5 months later,and two patients died.Conclusions Angiosarcoma is a rare and heterogeneous sarcoma with diverse clinical presentations.Local wide resection is the cornerstone of angiosarcoma treatment,and adjuvant therapy may improve its prognosis.

Objective To evaluate the effect of oxidative injury induced by peroxide oxidase on Klotho expression in mouse renal tubular epithelial cells (TCMK-1) and to explore the possible pathway.Methods TCMK-1 cells were exposed to H2O2 of different concentrations.Reactive oxygen species (ROS) was examined byflow cytometrry.Cell viability was assessed by CCK-8.Cell apoptosis was evaluated by flow cytometry and Hoechst 33258 staining.The expression of Klotho,apoptosis-associated proteins and anti-oxidant enzymes were determined by Western blotting.Results Compared with control group,after H2O2 stimulating TCMK-1 cell,ROS was dramatically elevated (all P ＜ 0.05) and the expression of anti-oxidant enzymes,SOD2 and CAT went down (all P ＜ 0.05);the expression of Klotho was inhibited (all P ＜ 0.05);cell viability of TCMK-1 cells was decreased (all P ＜ 0.05) in a dose-dependent manner (0.3 to 0.9 mmol/L);cell apoptosis was significantly increased in TCMK-1 cells following the concentration of H2O2 (all P ＜ 0.05);Bax/Bcl-2 and the phosphororation of JNK and p38 were obviously elevated in TCMK-1 by H2O2 induction (all P ＜ 0.05).Conclusion Oxidative injuries induced by H2O2 significantly suppresses the expression of Klotho in TCMK-1 cells.And cell apoptosis was increased,p38 and JNK pathway was activated.

Objective To investigate the relationship between the variation of endothelial progenitor cells (EPC) number and cardiovascular diseases (CVD) in maintenance hemodialysis (MHD) patients ,and discuss the function of EPC in the progression of CVD in MHD. Methods One hundred and fifteen MHD patients over 18 years whose dialysis vintage was over six months from Department of Nephrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine were enrolled. They were divided into CVD group and non ? CVD group by medical history, electrokardiographie (EKG), cardiac ultrasound, peripheral vascular imaging and cardiovascular imaging. Peripheral blood (5 ml) was collected for detecting EPC number by flow cytometry as CD34/CD133/vascular endothelial growth factor receptor 2 (VEGFR2) cells. The EPC number between CVD group and non?CVD group was compared. The relationship between the decrease of EPC number and CVD risks in MHD patients was analyzed by logistic regression analysis. In a three?year follow?up, the death and new CVD events of the two groups were compared in order to discuss the relationship between EPC number and adverse events. Results Among 115 MHD patients, the average age was 61.57 ± 12.76, male/female was 71/44, the average dialysis vintage was (86.24 ± 56.31) months, the average Kt/V was 1.69 ± 0.29 and average ultrafiltration volume was (2.48 ± 0.90) L. Forty?four patients in 115 (38.3%) were with concurrent CVD. The EPC number in CVD group was significantly lower than that in non CVD group (P=0.015). The CVD group had higher serum phosphate (P=0.013), higher glycosylated hemoglobin (P<0.001), but serum calcium, intact parathyroid hormone (iPTH) and other indicators had no significant difference between two groups. Multiple Logistic regression analysis showed that older age (OR=1.061), history of diabetes (OR=9.796), dialysis vintage (OR=1.015), serum phosphate (OR=3.766), decrease of EPC number (OR=0.909) were the independent impact factors of CVD events in MHD patients. There were 22 patients of the 115 MHD patients had encountered a new CVD event in a three?year follow?up between December 2012 and December 2015, 9 patients from the CVD group and 13 patients from the Non?CVD group, and there was no significant difference between two groups (P=0.776). Nine patients from the CVD group and 7 patients from the Non?CVD group died in the follow?up, and there was no significant difference (P=0.111). Seventy?one MHD patients from the non?CVD group were divided into two groups by the median of EPC number. There were 3 patients in the higher EPC number group encountered CVD events and 10 patients in the lower EPC number group encountered CVD events, which had significant difference (P=0.024). Conclusion The decrease of circulating EPC number may be related with CVD events in MHD patients. Even adjusted by age, sex, diabetes, dialysis vintage and serum phosphate, decreased EPC number is still the independent risk factor of CVD events in MHD patients. The decrease of EPC number in MHD patients may be used to predict the occurrence of cardiovascular events.

Objective To investigate the role of ATP￣binding cassette ( ABC ) family on the resistance of nasopharyngeal carcinoma (NPC) stem cells (CSCs) to cisplatin. Methods We compared the differences between the drug extravasation capability of CNE￣2 and CNE￣2S by using Rhodamine￣123 efflux assay. We determined the mRNA and protein expression levels of ABC transport family members, including ABCA3,ABCB1,ABCB5,ABCC1,ABCC2 and ABCG2,after 48 h being treated with 1 μmol.L-1 cisplatin by RT￣PC and Western blotting.Rhoamine￣123 efflux and apoptosis by cisplatin in two kinds of cells was examined by ABCA3 gene silencing with specific small￣interfering RNA. Results The IC50 of cisplatin on CNE￣2S was 4.1 fold to that on CNE￣2(P<0.05).For the relative drug effluent activity and Na+K+ ATPase activity,CNE￣2S was 4.8 fold to CNE￣2(P<0.05),suggested that CNE￣2S expressed more ABCA3,ABCB1,ABCC1 and ABCG2 in comparison to CNE￣2(P<0.05).After 48 h treatment with 1 μmol.L-1 cisplatin,ABCA3 specifically highly expressed in CNE￣2S (P<0.05), and knocking down of ABCA3 resulted in reduction of rhodamine￣123 efflux and increase of apoptosis. Conclusion The cisplatin resistance of NPC CSCs is associated with enhanced expression of ABCA3,ABCC1 and ABCG2, suppression of ABCA3 could reverse the resistance of NPC CSCs to cisplatin.