Objective To investigate the expression and significance of myeloid cell leukemia-1 (Mcl-1 )gene and protein in Hepatocellular Carcinoma(HCC).Methods The expression of Mcl-1 was detected respectively by Reverse Transcription-Polymerase Chain Reaction (RT-PCR),Western blot and ENVISION immunohistochemistry in 20 HCC specimens,19 liver cirrhosis(LC)specimens,and 12 control ones.Their relationship with clinical and pathological characteristics of HCC was investigated.Results The expression of Mcl-1 mRNA in the control group,LC group and HCC group was 0.52±0.17, 3.46±1.7,3.65±2.93,respectively.The level in HCC and LC group was statistically different compared with the control group,respectively (t=7.925,5.334,P<0.05).The relative expression of Mcl-1 protein in LC group (0.51±0.35)and HCC group (0.75±0.36)were significantly higher than that in the control group (0.21±0.19)(t=5.526,6.355,P<0.05).The positive expression rate of Mcl-1 in HCC group was 55.00% (11/20),significantly higher than that in the con-trol group 33.33% (4/12)(t=7.835,P<0.05).The positive expression of Mcl-1 was related to tumor necrosis and TNM staging (χ2=4.201,P<0.05).Conclusion Mcl-1 gene and protein are differentially expressed in HCC,LC and the control, which may be involved in the occurrence,development and malignant transformation of HCC.

Adolescent ; Angiography ; Child, Preschool ; Female ; Hemoptysis ; diagnostic imaging ; drug therapy ; Humans ; Male

In order to further study mouse embryonic stem cells(ES cells),lentiviral vector PLL-IRES-Nanog-Neo was constructed.Mouse ES cells overexpressed nanog by mediation of lentiviral were cultured on mouse fetal fibroblast feeders after 2 weeks under G418 media and examined according to gowth characteristics. Results were showed that 918 bp nanog fragments were expressed in mouse ES cells mediated by lentiviral vector PLL-IRES-Nanog-Neo,mouse nanog-ES cells were taken on mass-like image and positve with alkaline phosphatase staining and Oct4 and SSEA1 immunocytochemistry under no LIF condition in the media. It is concluded that mouse ES cells Elevated nanog gene expression by mediation of lentiviral were constucted and cultured.

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

OBJECTIVETo explore the cytotoxic responses of spleen T lymphocytes (CTL) in BALB/c mice induced by recombinant HSP110-HER2/neu ICD complex.

METHODSTumor-bearing mouse model was immunized by HSP110-HER2/neu ICD complex. The IFN-γ level secreted by activated spleen T lymphocytes was detected by enzyme linked immunospot assay (ELISPOT). The corresponding CTL activity was measured by granzyme release assay.

RESULTSThe BALB/c mouse model of human mammary tumor highly expressing HER2/neu was established. HSP110-HER2/neu ICD complex immunization led to a significantly higher level of INF-γ than that in HSP110-P(789-797) immunized and HER2/neu ICD immunized mice. HSP110-HER2/neu ICD complex immunized animals also show significant CTL activity. The results of immunohistochemical staining showed that the number of blue spots in the PBS group was 4.57 ± 1.33, HSP110 group 6.83 ± 2.08, HER2/neu ICD group 16.17 ± 2.86, HSP110-P(789-797) group 43.67 ± 4.78, and SP110-HER2/neu ICD group 76.51 ± 8.17. The number of IFN-γ-secreting spleen lymphocytes in the HSP110-HER2/neu ICD group was significantly higher than that in the HSP110-P(789-797) group, and that of HSP110-P(789-797) group was significantly higher than that of HER2/neu ICD group (P < 0.01). The target cell-killing rate of the PBS group was (8.15 ± 1.27)%, HSP110 group (9.51 ± 1.51)%, HER2/neu ICD group (14.03 ± 2.45)%, HSP110-P(789-797) group (25.99 ± 3.04)% and HSP110-HER2/neu ICD group (38.15 ± 3.95)% (all P < 0.01).

CONCLUSIONSHSP110-HER2/neu ICD complex can promote the proliferation and maturation of T lymphocytes into CTLs, and might be used as anti-tumor vaccine to induce potent cytotoxic T lymophocyte immunoresponse against specific tumor cells.

Animals ; Breast Neoplasms ; metabolism ; pathology ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Cell Proliferation ; Female ; HSP110 Heat-Shock Proteins ; immunology ; Humans ; Interferon-gamma ; metabolism ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Random Allocation ; Receptor, ErbB-2 ; immunology ; metabolism ; Recombinant Proteins ; immunology ; Spleen ; cytology ; immunology ; T-Lymphocytes ; cytology ; immunology ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, Synthetic ; immunology

Objective:To observe the clinical efficacy of point application with Da Huang (Rhizoma Rhei Cruda) powder at Shenque (CV 8) plus moxa-salt hot compress on the umbilicus for preventing gastrointestinal adverse reactions after chemotherapy for non-Hodgkin lymphoma (NHL).Methods:A total of 60 cases with NHL under chemotherapy were divided into two groups by hospitalization order,with 30 cases in each group.The control group was treated with routine nursing and the observation group was additionally given point application with Da Huang (Rhizoma Rhei Cruda) powder plus moxa-salt hot compress on the umbilicus,to compare the effect in preventing gastrointestinal adverse reactions after chemotherapy between the two groups.Results:The occurrence rates of constipation,nausea,vomiting and poor appetite on the second day and fifth day after chemotherapy were obviously lower in the observation group than those in the control group,with statistically significant differences between the two groups (all P＜0.05).Conclusion:The point application with Da Huang (Rhizoma Rhei Cruda) powder at Shenque (CV 8) plus maxa-salt hot compress on the umbilicus can produce more significant efficacy in preventing the gastrointestinal adverse reactions after chemotherapy for NHL than routine nursing.Moreover,it is simple and easy to be used and popularized.

Objective To investigate the suppression effects of Tripotolide (TL) on the pancreatic cancer xenograft models and angiogenesis. Methods The growth suppression effect of TL on SW1990 was determined using cell count kit (CCK-8), apoptotic cells induced by TL were examined by morphology and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. The inhibitory effects of TL on the growth of tumor xenografts and tumor microvascular density (MVD) were investigated. ResultsTL inhibited the growth and proliferation of SW1990 cells in a concentration-dependent and time-dependent manner. The inhibition ratios of cells treated at 160 mg/ml TL for 24 h was 50. 6%, the apoptotic rate increased from 9.6% in the control group to 45.1% (P <0.01 ). The inhibition rate of cancer xenograft growth was 89.9% when TL was intratumorally injected at the dose of 0.5 mg/kg. The expression of VEGF in tumor tissue decreased while MVD also decreased from 36.25±8.64 to 9.87±3.34 (P <0.01 ). ConclusionsTL induced prominent growth inhibition and apoptosis in human pancreatic cancer cell lines. TL.can attenuate the growth of pancreatic caner xenografts through its effect on antiangiogenesis.

Objective To explore the hurt of susceptibility organs and critical orgens followed by rotavirus (RV) infection of whole body in newborn mouse.Methods RV strain was derived from the stool samples of patients with RV diarrhea and was proved to be long type by methods of ELISA and polyacrylamide gel electrophoresis (PAGE). RV was inoculated by the pathways of taking orally and injected to abdominal cavities,respectively. The pathological changes of the newborn mouse model infected with human natural RV by light microscope and electron microscope. The gene probe was marked by digoxin.The direct prove of RV infection in these organs was got by the detection of in situ PCR. Results Pathological changes were found in the small intestinal villus,lamina propria of the stomach and the heart cells of the mice taken RV orally.The mice with intraabdominal RV injection showed pathological changes of the cells in the small intestinal villus,liver and kidneys observed by electron microscope.Shortened small intestinal villus,nuclear membrane disorganization,massive vacuolization,mitochondrial swelling and rough endoplasmic reticulum dilation were observed in the cells of small intestinal.In the liver of the mice,marked mitochondrial swelling and agglutination,cell nucleus pyknosis or collapse,presence of numerous lipid droplets and vacuoles were found in the li-ver cells,with lymphocyte and plasmacyte infiltration.Obvious dilatation and shedding of the microvillus were found in cholangioles.The mitochondria of the proximal convoluted renal tubule showed mild swelling,but the cells in the heart and lung did not display obvious changes.Conclusion RV can damage lots of extra intestinal organs of the newborn mice if RV diffuses to the whole body of the mice.

In order to explore the clinical hypolipidemic features of Daidai flavone extract, the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats were studied and compared. The study established the quantitative determination method of naringin and neohesperidin in plasma by UPLC-MS. Study compared the pharmacokinetics differences of naringin and noehesperidin in normal and hyperlipemia rats on the basis of establishment of hyperlipemia model. Results indicated that the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats showed significant differences. The C(max) of naringin and neohesperidin in hyperlipemia rats plasma after oral administration of Daidai flavone extract increased obviously, while t1/2, MRT and AUC0-24 h decreased, compared to normal rats. But t(max) showed no differences to that of normal rats. The results further proved Daidai flavone extract would have better hypolipidemic effect in the hyperlipemia pathological status. And the characteristic active ingredients naringin and noehesperidin were the material base of Daidai flavone extract to express the hypolipidemic effect.
Animals ; Citrus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Flavones ; chemistry ; Hyperlipidemias ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective To observe the clinical efficacy of Zhuang’s Moxibustion plus acupuncture in treating spastic paralysis due to craniocerebral injury.Method Ninety-two patients with spastic paralysis due to craniocerebral injury were randomized into a treatment group of 60 cases and a control group of 32 cases. The control group was intervened by conventional internal medicine and rehabilitation, while the treatment group was intervened by Zhuang’s moxibustion plus acupuncture in addition to the intervention given to the control group. The modified Ashworth Scale (MAS) was adopted to evaluate the clinical efficacy.Result The total effective rate was 75.0% in the treatment group versus 65.6% in the control group, and the between-group difference was statistically significant (P<0.01).Conclusion Zhuang’s moxibustion plus acupuncture is an effective approach in treating spastic paralysis due to craniocerebral injury.