The known members of inhibitor of growth (ING) gene family are considered as candidate tumor suppressor genes. ING4, a novel member of ING family, is recently reported to regulate brain tumour angiogenesis through transcriptional repression of NF-?B-responsive genes, induce G2/M arrest by the increased p21 expression in a p53-dependent manner, suppress the loss of contact inhibition and represses activation of the hypoxia inducible factor, which plays an important role in the progression of tumorigenesis. However, seldom studies about ING4 inducing tumor cells apoptosis were reported.The C6 cells (mouse glioma cells) were infected respectively with the blank adenovirus carrying GFP (Ad) and the recombinated Ad-hING4-His, then RT-PCR assay was used to detect the transcriptions of hING4, as well Western-blotting assay was ued to detect the expressions of hING4. The effects of hING4 expression upon C6 cells were observed, and the growth curve was drawed and tumor control rates were calculated. The C6 cells, which were affected by blank Ad and Ad-hING4-His, were respectively observed by LSCM (laser scan confocal microscope) and transmission electron microscope (TEM), detected by flow cytometry; and the genomic DNA of both groups were extracted and electrophoresised in agarose gel to examinate the DNA fragments. The results showed hING4 can significantly inhibit the growth of C6 cells by promoting the cell’s apoptosis, which probably is the first one to prove this property of ING4.The experimental and theoretical foundation for gene therapy for gliomas with ING4 in the future was established.

Objective To evaluate the clinical effectiveness in screening asymptomatic diabetic peripheral neuropathy(ADPN) by the Michigan neuropathy screening instrument (MNSI) and the Toronto clinical scoring system(TCSS).Methods MNSI,TCSS and neural electrophysiological test (NET) were conducted in 232 neurologically asymptomatic type 2 diabetes patients.By using the results of NET as the golden criteria for diagnosis of ADPN,we evaluated the effectiveness of the two different scoring system by the receiver operator characteristic curve.The sensitivity,specificity,positive and negative predictive values,accuracy,Youden indexes and kappa values on different diagnostic cut-off points of MNSI and TCSS were analyzed.The correlation between the two different scoring system and the risk factors of diabetic peripheral neuropathy (DPN) were also analyzed.Results The area under the ROC curve of MNSI and TCSS were 0.792,0.704,respectively.The sensitivity,specificity,accuracy,Youden indexes and kappa values of MNSI over 2 and TCSS over 2 were 66.2％ vs 73.3％,90.4％ vs 63.7％,78.3％ vs 68.5％,0.566 vs 0.370,and 0.588 vs 0.345,respectively.MNSI was better than TCSS in the effectiveness of diagnosing ADPN and consistence with the result of NET.Moreover,MNSI was associated with the most related risk factors of DPN including age,glycosylated hemoglobin (HbA1c),HbA1c × disease duration,islet function and HDL-C.Conclusions MNSI could be used as a relatively simple and reliable method for clinical and epidemiological screening and assessment of ADPN.

Objective:To examine the effects of chronic intermittent hypoxia(CIH)on the NF-κB,IL-6 and PGE2 level in rats with periodontitis.Methods:32 male SD rats(6 weeks old)were randomly divided into 4 groups(n =8),group A(normoxic control),B (normoxic periodontitis),C(CIH)and D(periodontitis +CIH).Periodontitis model was established in the upper second molars by liga-tion technique and high-glucose diet in the rats of group B and D.The rats in the group C and D were subjected to CIH in a cycle of al-ternative nitrogen and oxygen in a closed chamber.The chamber was filled with nadir and zenith ambient oxygen every 1 20 seconds per cycle for 8 hours per day.The rats were sacrificed and the gingival tissues were examined for the detection of IL-6 and PGE2 expression by ELISA,and NF-κB expression by immunohistochemistry.Results:Histology revealed apical migration of junctional epithetlium and crestal alveolar bone resorption in group B and D,and in the above phenomena of group D was the severest.The content of NF-κB,IL-6 and PGE2 in group B,C,D was higher than that in group A(P <0.05),and in group D was the highest(P <0.05).Conclusion:Chro-nic intermittent hypoxia can aggravate the inflammation of periodontitis.

The biological!activities i.e. antineoplastic activities and antiviral activity of the two novel kinds of interferons: hIFN-?1 and hIFN-? were studied and compared. First the fusion expression vector: pcDNA3.1A-hIFN-?1-His and pcDNA3.1A-hIFN-?-His by PCR was constructed,then the two kinds of plasmids were transfected into WI-38 (human embryonic lung cells) with liposome. And cytopathic effect (CPE) suppression test was used to study and compare the antiviral activities of rhIFN-?1-His and rhIFN-?-His, meanwhile MTT assay was used to detect their antineoplastic activities.It was found that, antiproliferative activity and MxA protein induction shown by rhIFN-?1-His is more powerful than of rhIFN-?-His. The antiviral molecular mechanisms of both hIFN-?1 and hIFN-? are related to MxA.The foundation for further study on the bioactivities and mechanism of action of hIFN-?1 and hIFN-? was established.

Objective To discuss the influence of eldepryl on the expressions of glial fibrillary acidic protein (GFAP) and cd11b in substantia nigra and striatum in the rats with Parkinson’s disease (PD),and to clarify the regulatory role of eldepryl in the gliacytes.Methods 72 SD rats were randomly divided into control group,PD model group and eldepryl group,and each group was divided randomly into 4 d and 8 d subgroups (n=12)after the success of model preparation.The PD rat models were established by injecting rotenone in subcutaneous.The number of GFAP and cd11b positive cells and the expressions of GFAP and cd11b were detected by immunohistochemistry and Western blotting method.Results The GFAP and cd11b positive cells were all in a resting state in control group, the GFAP-positive cell body was slender and irregular and had elongated protrusions;the cd1 1 b-positive cell body was small and branch-like,and it had more slender protrusions.The GFAP and cd11b positive cells were all in a active state in model group, the GFAP-positive cell body was hypertrophy, the proj ections increased thickening;the cd1 1 b-positive cell body was more bigger, the proj ections were shorter and thicker, and the number was increased.Compared with model group, the GFAP-positive cell body and protrusions were more slender, the CD11b-positive cell body was more smaller,the projections were more slender,and the number was decreased in eldepryl group.There were a small amount of expression of GFAP and cd11b positive cells in substantia nigra and striatum in the rats in control group,and there was no significant difference between 8 d group and 4 d group(P>0.05). The number of GFAP and cd11b positive cells and the protein expression levels were significantly increased in model group compared with control group(P<0.01);there was more expression in 8 d group compared with 4 d group,but there was no significant difference(P>0.05).The number of GFAP and cd11b positive cells and the protein expression levels in eldepryl group were significantly reduced compared with model group(P<0.01);there were less expression in 8 d group compared with 4 d group, and there was significant difference (P<0.05 ). Conclusion There are activation and proliferation of the gliacytes in substantia nigra and striatum in the rats with PD,and eldepryl can inhibit the activation and proliferation of gliacytes.

Objective To explore the clinical regularity of drugs on epidemic encephalitis B with text mining technique. Methods The data set on epidemic encephalitis B was downloaded from CBM database. The regularities of Chinese herbal medicine, Chinese patent medicine (CPM), western medicine and the combination of CPM and western medicine on epidemic encephalitis B were mined by data slicing algorithm. The results are showed visually with Cytoscape 2.8 software. Results The main function of herbal pieces was focused on clearing heat and removing toxicity. The herbal pieces such as Rehmanniae Radix Isatidis Raxis, Isatids Folium, Paeoniae Radis rubra and Scutellariae Radix were most often used. Angong Niuhuang Wan and Qingkailing Injection were usually used as CPM. For western medicine, antiviral drugs such as interferon and ribavirin were often used, and they were often used together with brain cells nutrition medicine, hormone, immunopotentiator and so on, and they were also frequently used together with CPM such as Angong Niuhuang Wan and Qingkailing Injection. Conclusion Text mining approach provides an important method in exploring the medication regularity for diseas, and provide refenrence for clinical use.

OBJECTIVETo observe the killing effect of Herceptin and adriamycin sequentially applied on breast cancer cell line in vitro.

METHODSBT-474 human breast cancer cells in exponential growth phase were treated with Herceptin alone, adriamycin alone and their sequential administration (Herceptin before adriamycin and vice versa), respectively. Under optical microscope, the morphological changes of the cells were observed before and after drug administration. The expression rate and mean fluorescence intensity (MFI) of HER-2/neu and cell death rate were detected by flow cytometry.

RESULTSMicroscopically, the cells treated with different protocols all exhibited such changes as darkening and increase of cellular debris with irregular cell morphology. Flow cytometry revealed no significant difference in the expression rate of HER-2/neu in each group before and after treatment, but the MFI of HER-2/neu and death rate of the treated cells were significant different from those of the control group (P<0.05). The cell death rate of Herceptin-pretreated cells was significantly higher than that of adriamycin-pretreated ones (P<0.05).

CONCLUSIONHerceptin pretreatment enhances the killing effect of adriamycin on breast cancer cell line BT-474, which provides experimental evidence for designing clinical sequential biochemotherapy of breast cancer.

Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Death ; drug effects ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Synergism ; Female ; Flow Cytometry ; Humans ; Receptor, ErbB-2 ; biosynthesis ; Trastuzumab

OBJECTIVETo observe the effect of recombinant adenovirus Ad-ING4 on K562 cells.

METHODSHuman ING4 recombinant transfer vector pAdTrack-CMV-ING4 was constructed by enzyme digest and ligation of human ING4 gene which was obtained through site specific point mutation of mouse ING4. The vector was co-transduced into BJ5183 E. coli with pAdEasy-1. The new recombinant adenovirus vector pAdEasy-1-pAdTrack-CMV-hING4 was transfected into QBI-293A cells. To obtain the ING4 recombined adenovirus (Ad-ING4). Ad-ING4 was used to infect K562 cells. The effect on K562 cells of ING4 was tested by LSCM FCM and immunohistochemistry.

RESULTSHuman ING4 recombinant adenovirus vector was constructed successfully, and high titre ING4 recombinant adenovirus (Ad-ING4) was obtained. ING4 can down-regulate the expression of bcl-2 and up-regulate expression of bax. The apoptosis of K562 cells induced by ING4 was proved by LSCM FCM and immunohistochemistry. The apoptosis rate was 19.7% (after 72h), which displayed significant difference compared with that of control groups (P < 0.01).

CONCLUSIONAd-ING4 can inhibit the growth of K562 cells and induce the cells apoptosis. The human ING4 recombinant adenoviral vector constructed might provide an approach to the target therapy of tumors.

Adenoviridae ; genetics ; Animals ; Apoptosis ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; Cell Cycle Proteins ; genetics ; Cell Proliferation ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; K562 Cells ; Mice ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Plasmids ; genetics ; Transfection ; Transformation, Bacterial ; Tumor Suppressor Proteins ; genetics

The human interleukin-17F(hIL-17F) gene was amplified by RT-PCR from PHA-activated human peripheral blood mononuclear cells (PBMCs). It was then subcloned into the retrovirus vector pSIV-1. The pSIV-1/hIL-17F together with its two-helper virus vectors pHIT456 and pHIT60 cotransfected into the package cell 293T by lipofectin to produce mature recombinant retrovirus, which was then used to infect SMMC-7721 hepatocarcinoma cells (HCCs), and the cells were selected in the presence of G418. The integration, transcription, expression of hIL-17F gene in SMMC-7721 cells was identified by PCR, RT-PCR and Western blot respectively. MTT and FCM showed that hIL-17F couldn't alter the proliferation and cell cycle of SMMC-7721 cells, but ELISA showed that it could down-regulate IL-6, IL-8 and VEGF expression. The effect of rhIL-17F supernatant on growth suppressing of ECV304 cells was observed by MTT. The experiment of human hepatocarcinoma xenograft tumor in nude mice showed that the formation and growth rates of hIL-17F-transgenic SMMC-7721 showed an obvious decline, and VEGF and CD34 expression and angiogenesis of the transgenic neoplasms was also evidently defined. hIL-17F can markedly inhibit the growth of human hepatocarcinoma xenograft tumor in nude mice by antiangiogenesis. This study provided an experimental evidence for further conducting tumor gene therapy by targeting vascularity and exploiting antiangiogenic novel medicine related to hIL-17F.
Animals ; Cell Line, Tumor ; Cell Proliferation ; Genetic Therapy ; Humans ; Interleukin-17 ; genetics ; Liver Neoplasms, Experimental ; pathology ; therapy ; Mice ; Mice, Nude ; Retroviridae ; genetics ; Vascular Endothelial Growth Factor A ; analysis ; Xenograft Model Antitumor Assays

The biological activities i. e. antineoplastic activities and antiviral activity of the two novel kinds of interferons: hIFN-λ1 and hIFN-ε were studied and compared. First the fusion expression vector: pcDNA3.1A-hIFN-λ1-His and pcDNA3.1A-hIFN-ε-His by PCR was constructed, then the two kinds of plasmids were transfected into WI-38 (human embryonic lung cells ) with liposome. And cytopathic effect (CPE) suppression test was used to study and compare the antiviral activities of rhIFNλ1-His and rhIFN-ε-His, meanwhile MTT assay was used to detect their antineoplastic activities. It was found that, antiproliferative activity and MxA protein induction shown by rhIFN-λ1-His is more powerful than of rhIFN-ε -His. The antiviral molecular mechanisms of both hIFN-λ1 and hIFN-ε are related to MxA. The foundation for further study on the bioactivities and mechanism of action of hIFN-λ1 and hIFN-ε was established.