Objective To explore the mRNA and protein expressions of zinc-finger protein 217(ZNF217)in cisplatin(DDP)-sensitive and DDP-resistant human ovarian cancer cell lines.Methods Six strains of ovarian cancer cells,including 3 DDP-sensitive strains(A2780,SKOV-3 and COC1)and 3 DDP-resistant strains(A2780-DDP-R,SKOV-3-DDP-R and COC1-DDP-R)were selected.The relative luminescence unit(RLU)of cells was determined with ATP assay,the IC50 value and resistance index(RI)of DDP-resistant cell lines were calculated.Intracellular localization of ZNF217 protein in the 6 ovarian tumor cell lines was detected by immunofluorescent cytochemistry.The expressions of ZNF217 mRNA and protein were determined by RT-PCR and Western blotting,respectively.Results The IC50 values of DDP to A2780 and A2780-DDP-R were 18.1?2.3mg/L and 47.9?3.8mg/L(P

Objective To investigate the hospital distribution and drug resistance of pathogens of venous catheter-related infec-tion,so as to provide basis for appropriate usage of antibiotics in clinic.Methods The culture and drug susceptibility results of 369 venous catheter were analyzed retrospectively from 2009 January to 2012 December.Results 161 strains of pathogens were detec-ted among 369 venous catheter,the positive rate was 43.6%.There were 83 strains of Gram positive bacteria,accounting for 51.6%,and Staphylococcus epidermidis had the highest positive rate.There were 63 strains of Gram negative bacteria,accounting for 39.1%,and Klebsiella pneumoniae and Bauman Acinetobacter were main isolated bacteria.There were 15 strains of Fungi,ac-counting for 9.3%,and Candida parapsilosis had the highest positive rate.There was none Gram positive bacteria resistant to van-comycin and linezolid.Carbapenems remained high activity against Gram negative bacteria.Conclusion Staphylococcus epidermidis and Klebsiella pneumoniae are mainly detected in venous catheter in the hospital infection.

OBJECTIVE To study the rate and the risk factors of hospital infection after hemopoietic stem cell transplantation and improve treatment strategy.METHODS The clinical data of 21 cases of hemopoietic stem cell transplantation were analyzed respectively in our hospital.RESULTS Hospital infection was found in 15 cases,the infection rate was 71.4%.Fifteen kinds of bacteria and 4 kinds of fungi were observed.The risk factors were aggressive operation,the abuse of glucocorticoid and antibiotics.CONCLUSIONS The patients have a high rate of hospital infection after hemopoietic stem cell transplantation and a poor prognosis because of hospital infection.To decrease the rate of hospital infection after hemopoietic stem cell transplantation,the whole environmental protection should be carried out except decreased aggressive operation and the correct use of glucocorticoid and antibiotics.

Objective To establish pulmonary acute lung injury(ALI)model in rats.Methods ALI model was induced in rats by intratracheal Escherichia coli injection[3 ml/kg,O111B4,(4.0～6.0)×1012 CFU/L].Mechanical ventilation was applied 12,24,36,48 and 72 h after Escherichia coli injection,PaO_2/FiO_2 and dynamic compliance were recorded,and the normal control group was also subjected to mechanical ventilation.After the experiment,lungs were fixed with formalin to perform pathological examination.Results At 12,24,36,48 and 72 h,the PaO_2/FiO_2 were(30.71±7.95)kPa,(21.66±5.34)kPa,(21.09±4.75)kPa,(25.01±8.78)kPa and(33.82±8.02)kPa,respectively,which were significantly lower than that in the normal control group(63.82±3.03)kPa(P＜0.01).At 12,24,36,48 and 72 h,the Cdyn were(4.23±0.13)ml/(kg·kPa),(4.19±0.96)ml/(kg·kPa),(4.28±0.69)ml/(kg·kPa),(4.44±0.62)ml/(kg·kPa)and(4.58±0.35)ml/(kg·kPa)respectively,which were significantly lower than that in the normal control group(8.16±0.78)mL/(kg·kPa)(P＜0.01).At 12,24,36,48 and 72,the percentage of ALI was 71.4%,100.0%,100.0%,83.3%and 57.1%respectively,and the percentage of ARDS was 28.6%,85.7%,83.3%,66.7%,14.3%respectively.As for pathological examinations,predominance of alveolar collapse,fibrinous exudates,alveolar wall edema and neutrophil recruitment into the alveolar space was observed.Hyaline membrane formation was found.At 72 h,inflammation was relieved.Conclusion We successfully established pulmonary ALI/ARDS model in rats induced by intratracheal Escherichia coli injection,and acquired some useful information by observing the lung function and morphological changes at different time points.

Objective To study the adrenal function in rats with pulmonary acute lung injury (ALI) induced by Escherichia coli (0111B4) . Methods ALI rat model was induced by intratracheal injection of E.coli (3 ml/kg,0111B4,(4.4-5.6) x 10~(12) CFU/L).ALI rats were then randomly divided into three groups,and each group had 10 rats.Mechanical ventilation was applied at three time points,6 hours,24 hours,and 36 hours after injection At each time point 8 rats were used as control with saline administered intratracheally.The plasma ACTH and corticosterone levels were measured after stimulated by 100 μg porcine ACTH.Results Compared with control group,the model group had a higher level of plasma ACTH at each time point (P < 0.01).The plasma ACTH level reached the peak at 24 hours.The model group had a higher level of plasma corticosterone at 6 hours (P < 0.01) and 24 hours (P <0.05),but had a lower level of plasma corticosterone at 36 hours (P < 0.05).The plasma corticosterone level reached the peak at 6 hours in model group,which was higher than 24 hours (P < 0.05).After stimulated by ACTH,the increased levels of corticosterone were lower in model group than those in control group (6 hours,P < 0.05; 24 hours and 36 hours,P < 0.01).Conclusions Adrenal dysfunction may occur at early stage of ALI in rats.With the disease developed,adrenal response to ACTH decreased.Low dose corticotrophin-stimulated test could evaluate adrenal function in rats with pulmonary ALI induced by Escherichia coli (O111B4).

Objective To study the value of flow cytometry in identifying metastatic CK positive and negative nonhematopoietic neoplasms in bone marrow. Methods Twenty-six cell lines representing ten epithelial neoplasms, one lymphoma cell line and one human T cell lymphoblast-like cell line were purchased from American Tissue Culture Collection. From July 2009 to June 2010, five nonhematopoietic neoplasms,fifteen hematopoietic neoplasms and fifteen control patients with complete remession after hematopoietic stem cell transplantation were collected in Beijing Daopei Hospital. Cryopreserved cell lines were thawed and cultured until they entered log phase. After permeabilization, cell lines were analyzed by staining with cytoplasmic CK-FITC antibody using four-color flow cytometer. The percent CK positivity was measured by comparing with negative control. Bone marrow samples were stained with membrane and cytoplasmic antibodies according to our routine methods. Based on lineage markers and blast markers as well as CK expression, the relevant hematopoietic diseases were diagnosed or excluded according to 2008 World Health Organization diagnosis standards. Results All epithelial neoplasm cell lines expressed CK, with average positive percentage 81.1%. All the lymphoid tumor cell lines didn't express CK. Two epithelial neoplasms were CK positive, 100. 0% in thyroid carcinoma and 98. 2% in lung carcinoma, respectively. Hematopoietic tumor and control samples didn't express CK. They expressed relevant hematopoietic markers, such as CD45 as well as lineage markers, or CD138 and cytoplasmic immunoglobulin light chain. Three nonepithelial nonhematopoietic neoplasms didn't express CK. CK positive or negative nonhematopoietic neoplasms didn't express hematopoietic markers such as CD45, HLA-ABC and HLA-DR DP DQ, as well as lineage specific markers. Besides, CK positive might be helpful to suggest epithelial origin. Conclusion Flow cytometry with hematopoietic markers and CK can effectively exclude hematopoietic tumor and identify metastatic CK positive and negative nonhematopoietic neoplasms in bone marrow.

Objective To explore the anti-inflammatory effect of antiflammin-2 (AF2) and recombinant peptide sequence 2(R2) on acute lung injury of mouse. To observe the expression of clara cell 16000 protein (CC16) and surfactant protein A (SP-A) in the lung of mouse inoculated with lipopolysaccharide (LPS) and the impact of AF2,R2,and glucocorticoids(hydrocortisone,HC) may have on the expression of the CC16 and SP-A in the lung of mice with acute lung injury. Methods Balb/c mice were inoculated with LPS (5 mg/kg) by intraperitoneal injection to set up ALI mice model. Mice weighed from 15 g to 16 g were grouped into control group, model group and treated groups respectively treated with AF2, R2 or HC. Mice in the control group were injected with physiological saline solution, while mice in the other four groups were inoculated with LPS to induce acute lung injury. Then animals in the treated groups were treated with AF2, R2 or HC each on a dose of 2 mg/kg also through intraperitoneal injection,while those of the control group and the model group, were given equivalent physiological saline solution as a placebo. The respiratory rate of all of these animals were recorded 6 hours after the injection. And at the time point of 12 hour,all the mice were sacrificed for a preparation of the whole lung tissue for the sake of a pathological investigation ,or for extractions of RNA for a semiquantitative analysis of the expression of CC16 and SP-A within the lungs. Results (1) An obvious attenuation of the respiratory rates of the three treated groups were observed when comparing with that of the mice in the model group without any anti-inflammatory treatment. (2) Remarkable extenuation of the extent of intra-alveolar and intersticial hemorrhage and infiltration of inflammatory cells were observed within the treated groups comparing with that of the model group. (3) An attenuate expressions of CC16 or SP-A were observed in the model group,while obvious uptrend of CC16 expression was observed in AF2 treated groups and increase of SP-A expressions were found in R2 and HC treated groups. Conclusion The anti-inflammatory effect of the peptide, AF-2 or R2, has been conformed on ALI mice model induced by LPS.

Background and purpose: Hepatocellular carcinoma (HCC) is a hypervascular tumor associated with a poor prognosis and lack of effective treatments. Consequently, identifying novel therapeutic strategies are urgently needed. We have previously shown that the kringle 1 domain of human hepatocyte growth factor (HGFK1) is a more effective anti-angiogenesis molecule than angiostatin. In this study, we observed the effects and mechanisms of HGFK1 gene on the HCC. Methods: A recombinant adeno-associated vires carrying the HGFK1 gene (rAAV-HGFK1) was constructed.HCC of rat was induced by McA-RH7777. rAAV-HGFK1 was used to treat the rat, median survival time and metastasis rate were observed. Results: Ten days after tumor cell inoculation, surgery were performed to confirm the tumor formation, PBS, rAAV-EGFP or rAAV-HGFK1 was injected directly into the tumor nodule followed by portal vein injection. Results from our study demonstrated that rAAV-HGFK1 treatment significantly prolonged the median survival time of the HCC bearing rats from 30 days (PBS and rAAV-EGFP groups) to 49 days (rAAV-HGFK1 group). More importantly rAAV-HGFK1 inhibited tumor growth and completely prevented liver, lung and peritoneal metastasis. In the controlled PBS and AAV-EGFP group, liver and peritoneal metastasis rate were both 100%, and lung metastasis rate was 100% and 83%, respectively. While there was no metastasis found in treatment group, with only 33% of ascites happened. This was most possibly due to the primary tumor in liver but not due to the metastasis. Moreover, at a higher magnification (1000×), it was clear that the HGFK1 protein was expressed mainly in the cytoplasma of liver cells. In parallel, IHC staining of CD31 also demonstrated a significantly lower level of microvessel density (MVD) (6.21±1.6) in the liver tumor of the AAV-HGFK1 treatment group, as compared to the two control PBS and AAV-EGFP groups (25.1±2.1 and 26.8±2.5, respectively, P<0.01). HE staining showed that AAV-HGFK1 treatment induced large areas of necrosis in the tumor tissues, while minimal areas of necrosis were observed in the tumor tissue in the control groups. In addition, no toxicity appeared when high dosage (4.8× 1012 vg/rat) of rAAV-HGFK1 was administered in rats. Conclusion: Results from this study demonstrated that HGFK1 inhibited the growth and metastasis of HCC and prolonged the survival time of animals with HCC through anti-angiogenesis effects. No obvious toxicity was observed. It might be the novel promising treatment for HCC and other cancers.

Objective: To study the relationship of LPPCN with neovascularization and to analyze its clini-copathologic significance, in an effort to find a new target for anti-vascular therapies. Methods: Sixty-eight ma-lignant melanoma specimens were analyzed to observe the distribution of LPPCN and to examine the expres-sion of CD105 and TGFβ1 using immunohistochemistry. The distribution of vasculogenic mimicry (VM) was observed by immunohistochemical and histochemical double staining of CD31 and PAS. Results: (1) The tu-lines and networks. Of the 68 cases of melanoma, 55.89% (38/68) were recognized as having LPPCN. (2) In malignant melanoma specimens, the rate of vasculogenic mimicry density (VMD) and microvessel density (MVD) labeled by CD105 in LPPCN-positive group were higher than those in LPPCN-negative group, with sig-nificant differences (P<0.05). VMD and MVD were positively-correlated with the density of LPPCN. The posi-tive expression of TGFβ1 in LPPCN-positive group was higher than that in LPPCN-negative group and its ex-pression in the regions of LPPCN was obviously higher than that in circumambient tumor cells, with a signifi-cant difference (P<0.05). The expression of TGFβ1 was positively correlated with MVD labeled by CD105. (3) There was no relationship between LPPCN and gender, age, site, tumor embolus, lymph node metastasis or distant metastasis (P>0.05), but LPPCN was correlated with tumor size, mitosis figure count and Breslow depth (P<0.05). Kaplan-Meier survival analysis showed the survival rate of patients with LPPCN was lower than that of patients without LPPCN, with a statistical significance (P<0.05). The presence of LPPCN indicat-ed poor prognosis. Conclusion: LPPCN exists in malignant melanoma and is associated with VM and angio- genesis. Some tumor cells undergoing LPPCN have a spacial foundation for VM and angiogenesis. LPPCN can be an index for the evaluation of the prognosis of melanoma patients.

This article was aimed to study the proteomic spectra expression of traditional Chinese medicine (TCM) cold and heat constitution rats with two-dimensional gel electrophoresis (2-DE), in order to search for the cold and heat-associated proteins for the investigation of the biological basis of TCM cold and heat body constitution formation. The total protein in rat’s liver cell was extracted. The 2-DE and MALDI-TOF/TOF mass spectrometry (MS) were used in the screening and identification of differentially expressed proteins of cold and heat constitution rats. The results showed that a total of 10 different points in the protein expression were obtained with statistical significance after screening and MS, which were carbamoyl-phosphate synthase, protein disulfide isomerase associated 3, catalase, hydroperoxide isomerase, cytosol aminopeptidase, glutamate dehydrogenase 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, heat shock protein 60 (Hsp60) precursor, homocysteine, 78 kDa glucose-regulated protein precursor. It was concluded that some differences were existed in the proteomic spectra expression of TCM cold and heat constitution rats. The abnormality of enzyme protein metabolism may be one of the material bases for the formation of cold and heat constitution.