Adult ; Colitis ; diagnosis ; surgery ; Cysts ; diagnosis ; surgery ; Female ; Humans ; Rectum ; pathology ; surgery

Objective To evaluate the safety and efficacy of neoadjuvant chemotherapy combined with PA-MSHA injection for the treatment of locally advanced breast cancer. Methods An open, randomized, controlled clinical trial was conducted in this study. 42 locally advanced breast cancer patients were randomly assigned to two groups, namely the experimental group (20 cases) and control group (22 cases). All the patients received chemotherapy of TEC regimen, while, in addition, the patients in experiment group received PA-MSHA injection. After the treatment, the efficacy of treatment was evaluated. The safety and tolerance of patients were also measured during the treatment. Results The overall response rate (CR+PR) [75.0 %(15/20)]in the experiment group was significant higher than that [54.6 %(12/22)]in control group (P < 0.01). Adverse reactions were found for 9 cases in experiment group, four of whom received medical care while the others recovered automatically. Conclusion PA-MSHA injection can significantly enhance the efficaey of neoadjuvant chemotherapy on the patients with locally advanced breast cancer. The PA-MSHA injection which has been proved safety in treatment is an ideal supplementary therapy for breast cancer.

BACKGROUNDSevere acute respiratory syndrome (SARS) is an infectious disease caused by SARS-CoV. There are no effective antiviral drugs for SARS although the epidemic of SARS was controlled. The aim of this study was to develop an RNAi (RNA interference) approach that specifically targeted the N gene sequence of severe acute respiratory syndrome associated coronavirus (SARS-CoV) by synthesizing short hairpin RNA (shRNA) in vivo, and to assess the inhibitory effect of this shRNA on SARS-CoV N antigen expression.

METHODSThe eukaryotic expression plasmid pEGFP-C1-N, containing SARS-CoV N gene, was co-transfected into 293 cells with either the RNAi plasmid pshRNA-N or unrelated control plasmid pshRNA-HBV-C4. At 24, 48 and 72 hours post transfection, the green fluorescence was observed through a fluorescence microscope. The RNA levels of SARS-CoV N were determined by reverse transcription polymerase chain reaction (RT-PCR). The expression of Green Fluorescent Protein (GFP) and protein N were detected using Western blot.

RESULTSThe vector, pshRNA-N expressing shRNA which targeted the N gene of SARS-CoV, was successfully constructed. The introduction of RNAi plasmid efficiently and specifically inhibited the synthesis of protein N. RT-PCR showed that RNAs of N gene were clearly reduced when the pEGFP-C1-N was cotransfected with pshRNA-N, whereas the control vector did not exhibit inhibitory effect on N gene transcription.

CONCLUSIONSOur results demonstrate that RNAi mediated silencing of SARS-CoV gene could effectively inhibit expression of SARS-CoV antigen, hence RNAi based strategy should be further explored as a more efficacious antiviral therapy of SARS-CoV infection.

Antigens, Viral ; genetics ; Cells, Cultured ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Nucleocapsid Proteins ; antagonists & inhibitors ; genetics ; RNA Interference ; SARS Virus ; genetics ; immunology ; Severe Acute Respiratory Syndrome ; therapy

OBJECTIVETo investigate the role of Twist in regulating the proliferation, migration, and invasion of osteosarcoma cells with different levels of malignancy.

METHODSThe baseline expressions of Twist in 3 different osteosarcoma cell lines (143B, MG63 and TE85) were detected using real-time PCR and Western blotting. The cells were infected with the recombinant adenoviruses Ad-Twist or Ad-siTwist for Twist overexpression or knockdown, respectively, and the cell growth curves were drawn to assess the cell proliferation. The migration abilities and invasiveness of the cells were evaluated using wound healing assay and Transwell assay. Luc-labeled 143B cells infected with Ad-Twist or Ad-siTwist were intrathecally injected to establish nude mouse models bearing osteosarcoma xenografts, in which the tumor formation was monitored using living body imaging technique.

RESULTSThe baseline expressions of Twist in the 3 osteosarcoma cells were significantly higher than that in C3H10 cells (P<0.05). Twist expression was the highest in 143B cells followed by MG63 cells, and was the lowest in TE85 cells, indicating its positive correlation with the level of malignancy of the osteosarcoma cells. Ad-Twist or Ad-siTwist infection efficiently enhanced or lowered Twist expressions at both mRNA and protein levels in osteosarcoma cells (P<0.05). Twist overexpression resulted in enhanced proliferation, migration and invasion abilities of osteosarcoma cells, and Twist knockdown obviously inhibited the cell proliferation, migration and invasion. In nude mice, 143B cells with Twist overexpression showed accelerated tumor formation compared with the control cells, while Twist knockdown significantly inhibited the tumor formation ability of the cells.

CONCLUSIONTwist overexpression can promote the proliferation, migration, invasion and tumorigenicity of osteosarcoma cells.

Objective To analyze the effects of S100A6 on Wnt/β-catenin signaling pathway and its molecular mechanism. Methods The expression of GST-hS10OA6 was induced with IPTG in Escherichia coli BL21, and the fusion protein was purified with glutathione-sepharose 4B beads. β-catenin level of human colon cancer cell line MG63 and human osteosarcoma cell line HCTl16 cells infected with AdS10OA6 was measured by Western blot. Luciferase activity assay was applied to analyze the effect of S100A6 on the β-catenin/TCF4 activity. The interactions between S100A6 and β-catenin/GSK-3β/Dvl/Axin were detected by GST-pulldown/Western blot. Results The β-catenin level in AdS100A6-infected MG63 and HCT116 cells was significantly increased in comparison with that in the AdGFP control group (P<0.01). The luciferase activity in human embryonic renal cell line 293 cells transfected with pTOP-Luc and followed by GST-hS100A6 treatment was increased by 20. 2-fold in comparison with that in the GST control group (P<0.01). The interaction between GST-hS100A6 and Axin was not found. Conclusion S100A6 up- regulates the Wnt/β-catenin signaling pathway, and this may be attributed to the interaction between S100A6 and β-catenin/GSK-3β/Dvl.

Objective To analyze the effects of S100A6 on Wnt/β-catenin signaling pathway and its molecular mechanism. Methods The expression of GST-hS10OA6 was induced with IPTG in Escherichia coli BL21, and the fusion protein was purified with glutathione-sepharose 4B beads. β-catenin level of human colon cancer cell line MG63 and human osteosarcoma cell line HCTl16 cells infected with AdS10OA6 was measured by Western blot. Luciferase activity assay was applied to analyze the effect of S100A6 on the β-catenin/TCF4 activity. The interactions between S100A6 and β-catenin/GSK-3β/Dvl/Axin were detected by GST-pulldown/Western blot. Results The β-catenin level in AdS100A6-infected MG63 and HCT116 cells was significantly increased in comparison with that in the AdGFP control group (P<0.01). The luciferase activity in human embryonic renal cell line 293 cells transfected with pTOP-Luc and followed by GST-hS100A6 treatment was increased by 20. 2-fold in comparison with that in the GST control group (P<0.01). The interaction between GST-hS100A6 and Axin was not found. Conclusion S100A6 up- regulates the Wnt/β-catenin signaling pathway, and this may be attributed to the interaction between S100A6 and β-catenin/GSK-3β/Dvl.

OBJECTIVESTo observe the inhibition of HBV replication and antigen expression by RNA interference aimed at different parts of the HBV genome.

METHODSFollowing the rules of shRNA expression vector design and construction, we constructed seven kinds of sequence specific vectors and two kinds of mutant shRNA expression ones. We then cotransfected those shRNA and HBV expression vectors into HepG2 cells using lipofectamine2000. The level of HBV replication was investigated using Southern blot and the antigen expression using ELISA.

RESULTSThe replication of HBV DNA was inhibited by many shRNAs, especially the ones against P1, S2, C2, S1 and X. The inhibition rate against P1 was as high as 95%. Results obtained with ELISA showed that the shRNAs targeting C2, C1 and S2 had high rates of inhibition to HBsAg.

CONCLUSIONThe replication and antigen expression of HBV could be inhibited by shRNAs aimed at four different open read frames, and higher inhibition rates of HBV replication and surface antigen expression could be obtained by P1 and C2, respectively.

Gene Expression ; Hep G2 Cells ; Hepatitis B virus ; genetics ; metabolism ; physiology ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Virus Replication

OBJECTIVETo study the influence of serum from scalded rats on the cytoskeleton of colonic smooth muscle cells (CSMC) of rats cultured in vitro, and to probe the possible mechanism of gastrointestinal motility disorder after burn.

METHODSCSMC isolated from healthy adult Wistar rat were cultured and divided into scald serum group (SS) and normal serum group (NS) according to the random number talbi. Two normal Wistar rats were used, one of which was inflicted with deep partial-thickness scald. Serum was obtained from blood collected from these two rats respectively and diluted to 20% in concentration. Serum from scald and normal rats were respectively added to the culture of CSMC in SS and NS groups. The expression of actin and the relative content of β-tubulin in CSMC was respectively determined with flow cytometry and Western blot at post treatment hour (PTH) 1, 3, 6, and 12 (with 10 samples in each group at each time point). Data were processed with t test.

RESULTSFluorescence intensity of actin in SS group at PTH 1, 3, 6, and 12 was respectively 59 ± 4, 26 ± 6, 39 ± 6, and 42 ± 6, all significantly lower than those in NS group (95 ± 10, 91 ± 10, 102 ± 9, and 97 ± 9, with t value respectively 10.528, 18.069, 18.748, 16.647, P < 0.05 or P < 0.01). In SS group, the fluorescence intensity decreased to the nadir at PTH 3, and then increased persistently at PTH 6 and 12. (2) Relative content of β-tubulin in SS group at PTH 1, 3, 6, and 12 was respectively 14.44 ± 0.26, 8.61 ± 0.19, 11.76 ± 0.31, and 12.13 ± 0.29, all significantly less than those in NS group (22.37 ± 1.15, 21.87 ± 1.79, 23.24 ± 1.55, and 21.99 ± 2.02, with t value respectively 21.176, 23.365, 23.000, 15.273, P values all below 0.01). In SS group, the relative content of β-tubulin decreased to the nadir at PTH 3 and increased slowly at PTH 6 and 12.

CONCLUSIONSThe reduction of CMSC content which has the tendency of increasing later, can be attributed to the influence of scald serum in initial stage. This may be related to the tolerance and adaptation to scald serum and self-repair of CMSC.

Animals ; Burns ; metabolism ; Cells, Cultured ; Colon ; cytology ; Cytoskeleton ; metabolism ; Male ; Microtubules ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Rats ; Rats, Wistar ; Serum

OBJECTIVE Using metabolomics to explore the essence of damp heat accumulation syndrome of infantile human cytomegalovirus hepatitis.METHODS 20 infantile HCMV hepatitis patients with damp heat accumulation syndrome and 20 healthy infants were recruited for investigation.Plasma and urine samples were analyzed using ultra-high-performance liquid chromatography-linear trap quadruple/orbitrap mass spectrometry (UHPLC-LTQ/Orbitrap-MS) to determine the alterations in metabolomic profiles of this syndrome.Orthogonal partial least squares-discriminate analysis (OPLS-DA) was applied to analyze the UHPLC-MS data obtained from these samples.The specificity and sensitivity of potential biomarkers were assessed according to the area under the receiver operator characteristic (ROC) curves.RESULTS The metabolomic analysis yielded 1076 plasma compounds and 414 urine compounds.Among them,22 plasma and 7 urine differential metabolites relating to sphingolipid,glycerophospholipid,histidine,glycerolipid,and fatty acid metabolism were identified.Sphingomyelin and triglycerides were screened as potential biomarkers and showed excellent discriminant performance.CONCLUSION This research can provide insights into the essence of HCMV hepatitis damp heat accumulation syndrome.

BACKGROUNDThe imaging evaluation of pain in patients who have had a hip arthroplasty (HA) is challenging, and traditional imaging techniques, including magnetic resonance imaging (MRI) and computerized tomography (CT), are limited by metallic artifact. The purpose of the present study was to investigate the use of modified MRI techniques to visualize periprosthetic soft tissues and the bone-implant interface, and to evaluate the value of MRI for the assessment of patients with painful hip arthroplasty.

METHODSFifty-six painful hips in fifty-six patients following primary HA were assessed using optimized MRI, CT and standardized radiographs. The diagnosis of MRI was correlated with intraoperative findings as well as with microbiological and histological examinations (when available). The sensitivity and the specificity of MRI diagnosis were determined according to final diagnosis. The chi-square test was performed to detect a difference between MRI and final diagnosis.

RESULTSForty-eight patients have received revision surgery and final diagnosis were established. MRI was demonstrated high sensitivity and specificity in detecting aseptic loosening (93% and 95%), periprosthetic infection (94% and 97%), adverse local tissue reaction (100% and 100%) and periprosthetic fracture (100% and 100%). MRI was determined to be the most sensitive technique in detecting implant loosening for any reason, with a sensitivity of 93.8% for acetabular shell and 97.1% for femoral stem, compared to 81.3% and 80.0% on CT, 75.0% and 77.1% on radiographs.

CONCLUSIONSOptimized MRI was effective for the assessment of the periprosthetic soft tissues and bone. The use of modified magnetic resonance imaging parameters provided a useful adjunct to conventional examinations for the evaluation of patients with painful hip arthroplasty.

Arthroplasty, Replacement, Hip ; adverse effects ; Hip Prosthesis ; adverse effects ; Humans ; Magnetic Resonance Imaging ; methods ; Pain ; diagnosis ; etiology ; Prospective Studies