Objective: The technique of isobaric tags for relative and absolute quantitation (iTRAQ) was used to analyze the expression levels of serum biomarkers of hepatocellular carcinoma (HCC) based on mass spectrometry. Methods: The literature about HCC biomarkers identified by screening of differential protein expression combined with matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) published between January 2003 and August 2010 were selected by searching PubMed database. Screening for serum differential proteins was performed in 20 HCC patients and 20 healthy volunteers by using iTRAQ labeling and MALDI-TOF-MS. The reported data of HCC biomarkers were compared with the screening results of serum differential protein expression in HCC. Results: Twenty-two pieces of eligible literature were obtained by information retrieval, involving 262 HCC biomarkers. Fifty-one proteins were identified having differential expression by using screening of serum differential protein expression in HCC. In the 51 proteins, the expression level of 23 proteins was high, while the expression level of 28 proteins was low. Eight differential proteins which were identified by screening, were in accordance with literature reports, and 6 differential proteins (alpha-1-antitrypsin, apolipoprotein E, ceruloplasmin, alpha fetoprotein, complement C3 and serum amyloid P-component) had the similar expression levels in accordance with literature reports. Conclusion: Six differential proteins identified in serum are the same as reported in the literatures, and these proteins may become potential HCC biomarkers. There is a need for further study to explore these differential proteins. Copyright© 2011 by Tumor.

To understand molecular modules related to polyunsaturated fatty acids (PUFA) synthesis and eventually produce PUFA at high efficiency, we developed a protein complex analysis technology in Synechocystis sp. PCC 6803, and applied it to identify possible partner proteins interacting with the key enzymes that catalyze PUFA biosynthesis. We first constructed a recombinant expression of protein of slr1609 encoding the fatty acid activation enzyme, by fusing 3xFLAG tag with the target protein. Then we verified its expression by Western blotting targeting 3xFLAG tag. To maximize purification of Slr1609 protein complex, we optimized the protein expression conditions of Slr1609 in Synechocystis in a 5 L fermenter by monitoring its gene expression using RT-qPCR. The purification of the Slr1609 protein complexes was demonstrated by a Native-PAGE analysis. Finally, LC-MS/MS proteomic analysis allowed identification of the possible partner proteins interacting with Slr1609.
Bacterial Proteins ; chemistry ; Chromatography, Liquid ; Fatty Acid Synthases ; chemistry ; Fatty Acids, Unsaturated ; biosynthesis ; Proteome ; chemistry ; Proteomics ; Synechocystis ; enzymology ; Tandem Mass Spectrometry

The purpose of the study is to construct R8 peptide (RRRRRRRR) and pH sensitive polyethylene glycols (PEG) co-modified liposomes (Cl-Lip) and utilize them in breast cancer treatment. The co-modified liposomes were prepared with soybean phospholipid, cholesterol, DSPE-PEG2K-R8 and PEG5K-Hz-PE (pH sensitive PEG). The size and zeta potential of Cl-Lip were also characterized. The in vitro experiment demonstrated that the Cl-Lip had high serum stability in 50% fetal bovine serum. The cellular uptake of Cl-Lip under different pre-incubated conditions was evaluated on 4T1 cells. And the endocytosis pathway, lysosome escape ability and tumor spheroid penetration ability were also evaluated. The results showed the particle size of the Cl-Lip was (110.4 ± 5.2) nm, PDI of the Cl-Lip was 0.207 ± 0.039 and zeta potential of the Cl-Lip was (-3.46 ± 0.05) mV. The cellular uptake of Cl-Lip on 4T1 cells was pH sensitive, as the cellular uptake of Cl-Lip pre-incubated in pH 6.0 was higher than that of pH 7.4 under each time point. The main endocytosis pathways of Cl-Lip under pH 6.0 were micropinocytosis and energy-dependent pathway. At the same time, the Cl-Lip with pre-incubation in pH 6.0 had high lysosome escape ability and high tumor spheroid penetration ability. All the above results demonstrated that the Cl-Lip we constructed had high pH sensitivity and is a promising drug delivery system.
Animals ; Cell Line, Tumor ; Cell-Penetrating Peptides ; chemical synthesis ; chemistry ; Cholesterol ; chemistry ; Drug Delivery Systems ; Liposomes ; Mice ; Oligopeptides ; chemical synthesis ; chemistry ; Particle Size ; Phospholipids ; chemistry ; Polyethylene Glycols

Objective To assess the value of combination of susceptibility weighted imaging (SWI)and dynamic susceptibility-weighted contrast-enhanced (DSC) perfusion-weighted magnetic resonance imaging in astrocytoma grading.Methods SWI and DSC scans were performed in 82 patients with pathologically confirmed astrocytoma.The patient group consisted of grade Ⅱ (15),grade Ⅲ (10),and grade Ⅳ (57).The intratumoral susceptibility signal intensity (ITSS) and relative cerebral blood volume (rCBV) max were used to determine the grade of astrocytomas by Kruskal Wallis test,Welch test,Spearman correlation coefficients,Pearson correlation coefficients,and receiver operating characteristic curve (ROC)statistic methods.Results There were no ITSS in 14 cases of low-grade astrocytomas,the degree of ITSS were grade 1 to 3 in anaplastic astrocytomas,the degree of ITSS were grade 3 in all of the glioblastomas,the degree of ITSS were significant difference in all grades (H =71.96,P ＜ 0.01).rCBV max in grade Ⅱ,grade Ⅲ and grade Ⅳ astrocytomas were 1.26 ± 0.42,3.59 ± 2.09 and 8.34 ± 1.16 respectively,rCBV max were significant difference in all grades (F' =681.72,P ＜ 0.01).ITSS showed significant correlation with rCBV max (r =0.72,P ＜0.01) and tumor grades (r =0.89,P ＜0.01),and rCBV and tumor grades showed significant correlation (r =0.78,P ＜ 0.01).Area under the ROC curve application SWI,DSC,SWI and DSC in differentiation of the grade Ⅱ and grade Ⅲ astrocytomas were 0.99,0.93,1.00,differentiate grade Ⅲ from grade Ⅳ were 0.70,0.94,0.94,and differentiate high-grade from low-grade astrocytomas were 1.00,0.99,1.00.Conclusions ITSS is helpful to determine the grade of astrocytomas.The use of SWI in combination with DSC may improve the diagnostic accuracy of astrocytoma grading.

Objective To explore the effect of electroacupuncture at Hegu (LI 4) on the expressions of P2X2 and P2X3 receptors in experimental pulpalgia rats.Method Forty-two male SD rats were randomized into a normal group (group N), a control group (group C), a pulpalgia model group (group M), an antagonist group (group A), an electroacupuncture group (group E), and an antagonist+electroacupuncture group (group AE), 7 rats in each group. Group N didn’t receive any interventions; group C received injection of normal saline into pulp cavity of the same dose as the injection in group M, and the cavity was then blocked by dental fillings 5-6 min later; in group M, maxillary first and second molar teeth were drilled (drill bit of 1 mm in diameter) to expose pulp and lipopolysaccharide (LPS) solution 5μg/μL was injected into the holes (1~3μL for each hole), and the holes were then covered by dental fillings 5-6 min later; group A received the same modeling method as that in group M, but A-317491 was injected together with LPS (0.5 mg/kg); group E received electroacupuncture at bilateral Hegu (LI 4) with needles retained for 30 min, once a day, totally for 3 times; group AE received the same electroacupuncture intervention after receiving the same treatments as that in group A. The rats’ behaviors and weight were observed for 30 min after intervention each day. The rats were sacrificed on the 4th day, and the mRNA expressions of P2X2 and P2X3 receptors andβ-actin in trigeminal ganglion were detected by using RT-PCR. The mRNA expressions were then compared among the groups.Result The behavioral changes in group M, E, and AE were more significant than that in group C and N (P<0.01); the behavioral changes in group A, E, and AE were less significant than that in group M (P<0.01). The weight in group C was significantly lower than that in group N (P<0.01); the weights in group M, A, E and AE were significantly lower than that in group C and N (P<0.01); the weights in group E and AE were significantly higher than that in group M and A (P<0.01,P<0.05); the weight in group A was slightly higher than that in group M (P<0.05); the weights in group AE was significantly higher than that in group E (P<0.01). The mRNA expressions of P2X2 receptor in group M, A, and AE were significantly higher than that in group N and C (P<0.01); the mRNA expressions of P2X2 receptor in group A, E, and AE were lower than that in group M (P<0.05); the mRNA expression of P2X2 receptor in group A was lower than that in group E (P<0.05); the mRNA expression of P2X2 receptor in group E was higher than that in group AE (P<0.05). The mRNA expressions of P2X3 receptor in group M, A, and E were significantly higher than that in group C (P<0.05) and group N (P<0.01); the mRNA expressions of P2X3 receptor in group A, E, and AE were significantly lower than that in group M (P<0.01); the mRNA expression of P2X3 receptor in group AE was markedly lower than that in group E (P<0.01).Conclusion The expressions of P2X2 and P2X3 receptors in trigeminal ganglion were increased in LPS-induced pulpalgia rats. Electroacupuncture at Hegu (LI 4) and injection of A-317491 both can down-regulate the mRNA expressions of P2X2 and P2X3 receptors, which is plausibly the action mechanism of electroacupuncture at Hegu (LI 4) in analgesia.

Animalcule Identification and Drug Susceptivity System is designed with virtual instrument technology. The paper introduces main functions and the special feature of the software design. It discusses how to accomplish database operations and serial communication with LabVIEW and achieve remote distribution of data with DataSocket tchnology.

OBJECTIVETo study the effects of dihydroartemisinin on the apoptosis of and the vascular endothelial growth factor (VEGF) expression in prostate cancer cell line PC-3M in androgen-independent prostate cancer.

METHODSPC-3M cells were treated with different doses (0, 25, 50 and 100 micromol/L) of dihydroartemisinin for 48 hours, their growth activity analyzed by MTT colorimetric assay and flow cytometry, and changes in the activities of caspase-3 and -8 detected by colorimetric assay. The expression of VEGF mRNA was determined by semi-quantitative RT-PCR, and that of the VEGF protein by Western blotting.

RESULTSCompared with the 0 micromol/L control group, the 25, 50 and 100 micromol/L dihydroartemisinin groups showed significantly increased apoptosis of PC-3M cells ([2.92 +/- 0.45]% vs [8.85 +/- 0.74]%, [12.83 +/- 0.84]% and [18.65 +/- 1.24]%, P < 0.01), and dose-dependent increase in the activities of caspase-8 ([0.47 +/- 0.05 ] U/microg vs [1.22 +/- 0.15], [1.76 +/- 0.07] and [2.91 +/- 0.24] U/microg, P < 0.01) and caspase-3 ([0.44 +/- 0.07] U/microg vs [0.95 +/- 0.08], [1.48 +/- 0.14] and [2.92 +/- 0.45] U/microg, P < 0.01). The expressions of VEGF mRNA and protein were decreased in a concentration-dependent manner.

CONCLUSIONDihydroartemisinin can significantly suppress the growth of PC-3M cells, promote their apoptosis and reduce the expressions of VEGF mRNA and protein, which may serve to explain its inhibitory effect on tumor and angiogenesis.

Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo improve the resectable rate of massive hepatic tumors and operative tolerance of hepatectomy in the treatment of advanced liver cancers.

METHODSSixteen cases of massive hepatic tumors were reviewed. The selective exclusion of hepatic outflow and inflow in hepatectomy was discussed.

RESULTSAll the patients had normal course after the operative procedure and no hepatic coma or other severe hepatic disturbances were observed.

CONCLUSIONWhile the selective exclusion of hepatic outflow and inflow were applied, the resectable rate of massive hepatic tumors and operative tolerance of hepatectomy were improved.

Adult ; Carcinoma, Hepatocellular ; physiopathology ; surgery ; Female ; Hemangioma, Cavernous ; physiopathology ; surgery ; Hepatectomy ; methods ; Hepatic Artery ; surgery ; Hepatic Veins ; surgery ; Humans ; Liver Circulation ; Liver Neoplasms ; physiopathology ; surgery ; Male ; Middle Aged

Objective To investigate the expression and activation of extracellular signal-regulated kinase 1/2 (ERK 1/2), its upstream molecule, epidermal growth factor receptor (EGFR), and downstream transcription factor, Ets-like protein 1 (ELK-1), in lesions of psoriasis vulgaris, and to evaluate the relationship between ERK pathway and psoriasis vulgaris. Methods Tissue samples were obtained from the lesions of 40 patients with psoriasis vulgaris and normal skin of 20 normal human controls. Immunohistochemistry and Western blot were performed to detect the expressions of phosphorylated ERK1/2, EGFR and ELK-1 in the tissue samples.Results As immunohistochemistry showed, the integrated optical density (IOD) of p-ERK1/2, p-EGFR and p-ELK-1 was 269.85 ± 57.96, 136.88 ± 30.33 and 237.61 ± 56.29 respectively in the psoriatic lesions, significantly higher than that in the normal controls ( 140.24 ± 24.42, 110.66 ± 28.99 and 119.04 ± 21.99, respectively, all P ＜ 0.05). A positive correlation was observed between the expression of p-EGFR and p-ERK1/2(r = 0.57, P ＜ 0.05) and between that of p-ERK1/2 and p-ELK-1 (r=0.72,P＜0.05) in psoriatic lesions.Conclusion The enhanced signal transduction through phosphorylated EGFR→ERK1/2→ELK-1 pathway may play a certain role in the pathophysiological process of psoriasis vulgaris.

OBJECTIVESTo investigate the clinical effect of sequential therapeutic intervention Yupei Qisun [compensating for weakness by invigorating Kidney (Shen) and Spleen (Pi) in advance] in Chinese medicine (CM) and hysteroscopic endometrial mechanical stimulation on the treatment of infertile patients with repeated implantation failure (RIF); and to study the differences in patients' endometrial thickness and type on the day of embryo transfer, serum hormone levels on embryo transfer day and clinical pregnancy outcomes.

METHODSIn the clinical study, 168 frozen-thawed embryo transfer (FET) cycles for couples with RIF conforming to the research protocol were randomly divided into three groups: a CM group with 56 cycles (CM combined with FET), a hysteroscopy group with 55 cycles (hysteroscopic endometrial mechanical stimulation), and a control group with 57 cycles (conventional FET). Differences in endometrial thickness on the embryo transfer day, levels of serum estradiol (E2) and progesterone (P) on the embryo transfer day, the E2/P ratio on the embryo transfer day, biochemical and clinical pregnancy rates, implantation rate, abnormal pregnancy rate and other indices were compared among the three groups.

RESULTSEndometrial thickness, E2 and P levels, and the E2/P ratio on embryo transfer day and other factors had no significant differences among groups. The biochemical pregnancy, clinical pregnancy, and implantation rates of the CM and hysteroscopy groups were significantly higher than the control group (P<0.05), and there were no significant differences between these two groups. The abnormal pregnancy rate had no significant difference among the three groups.

CONCLUSIONSSequential therapy of Yupei Qisun could significantly improve the clinical outcomes of rif-fet cycles, being equivalent to hysteroscopic endometrial mechanical stimulation, and provided a reliable method to treat such infertile couples.

Abortion, Habitual ; therapy ; Adult ; Embryo Implantation ; Embryo Loss ; therapy ; Embryo Transfer ; Endometrium ; pathology ; physiopathology ; Female ; Humans ; Hysteroscopy ; Infertility, Female ; pathology ; therapy ; Medicine, Chinese Traditional ; methods ; Physical Stimulation ; methods ; Pregnancy ; Retreatment ; statistics & numerical data