Objective To investigate the possible effects on radiosensitivity in human umbilical vein endothelial cells after transfection of pcDNA3.1+Ape1 plasmid. Methods The expressing vector pcDNA3.1+ Ape1, the control vector pcDNA3.1+or non-transfection cells was irradiated by 2, 4, 6, and 8 Gy photon beam at 48 h post-transfection. The value of initial and residual Oliver tail moment (OTM) under the alkaline single cell gelelectrophoresis assay and the colony forming test were utilized as the markers for the evaluaton of cells intrinsic radiosensitivity. The effect on radiosensitivity in human umbilical vein endothelial cells after transfection of the expressing vector pcDNA3.1+Ape1 was analyzed according to the radio-dose, compared to the empty vecor control and non-transfection cells. Results The initial and residual OTM value of endothelial cells transfected by 3 μg pcDNA3.1+Ape1 plasmid was lower significantly than ones of endothelial cells untransfected at 2 Gy irradiation (P<0.01), but was no significant difference at 8 Gy (P>0.05), and SF2 was higher remarkably in transfected cells than one in untransfected cells (P<0.05), but SF4, SF6 and SF8 were no significant differences (all of P>0.05). Conclusions The transfection of pcDNA3.1+Ape1 plasmid could enhance radioresistance of endothelial cells to the low-dose irradiation.

Objective To investigate the case of left-semicolon cancer with intestinal obstruction for the methods of one-stage resection and anastomosis.Methods The clinical data of ten patients with left-semicolon can- cer with intestinal obstruction treated by transanus ilues tube,were restrospectively analyzed from October 2004 to December 2006.Results No postoperative anastormotic leakage was found and the patients were clinically cured. Conclusion The technical problem that left-semicolon cancer with obstruction for one-stage resection and anastomo- sis was resolved by the application of transanus ilues tube.

ObjectiveTo observe the effect of community-based rehabilitation (CBR) on older stroke patients in ability of activities of daily living (ADL).Methods50 older stroke patients were randomly divided into the rehabilitation group and control group. The rehabilitation group was treated with motor function exercise and ADL training, while the control group only took medicine. Two groups were evaluated with Barthel index before and after treatment. ResultsScores of Barthel index on the rehabilitation group were higher than that on the control group after treatment, and there was a significantly difference between two groups (P<0.01). Conclusions CBR has the significant effect on improving ADL in older stroke patients.

OBJECTIVETo discuss the changes in Wnt pathway inhibiting factors in esophageal precancerosis lesions induced by methyl benzyl nitrosamine (MBNA) and the effect of Gexia Zhuyu decoction.

METHODWistar rats were subcutaneously injected with MBNA (3.5 mg x kg(-1) for twice per week to establish the model. Since the 1st day after the model establishment, they were orally administered with Gexia Zhuyu decoction (16, 8 mg x kg(-1)). At the 10th week, esophageal tissues were collected to observe the pathological changes of esophageal mucosa, detect SFRP1, sFRP4, Axin1, Axin2 and GSK-3β mRNA levels.by fluorescent quantitation PCR analysis and β-catenin protein level by Western blotting.

RESULTBeing induced by MBNA, rats in the model group showed slight atypical hyperplasia in the histopathological examination. Compared with the normal group, Gexia Zhuyu decoction dose high and low groups showed no significant pathomorphological and histological changes. The model group showed lower gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and higher β-catenin protein expression level (P < 0.01) than the normal control group. The Gexia Zhuyu decoction low dose group showed higher gene transcription levels of esophageal tissues sFRP1, sFRP4, Axin1 and Axin2 (P < 0.05 or P < 0.01) and lower β-catenin protein expression level (P < 0.01) than the normal control group.

CONCLUSIONUp-regulated β-catenin protein level and down-regulated Wnt pathway could enhance Wnt pathway activity of MBNA-induced esophageal precancerous lesions. Gexia Zhuyu decoction could down-regulate the β-catenin protein level and up-regulate the transcription level of Wnt pathway inhibiting factors, but could not block MBNA-induced esophageal precancerosis lesions.

Animals ; Axin Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Esophageal Diseases ; drug therapy ; genetics ; metabolism ; pathology ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Male ; Necrosis ; Nitrosamines ; adverse effects ; Proteins ; genetics ; metabolism ; Rats ; Rats, Wistar ; Wnt Proteins ; genetics ; metabolism ; Wnt Signaling Pathway ; drug effects

Molecular methods and fluoroscopic techniques suggest that rich microbial diversity exist in the marine environment, but less than 1% of these microbes can be cultured in the laboratory conditions, and that the cultivable dominant species were even less. This limitation has long been a barrier to the development of environmental microbiology and the utilization of marine resources. In the past decade, novel methods for culture and detection of these uncultured marine microbes have successfully applied to obtain several conventionally-uncultured microbes including those from extreme environments. Those progresses have inspired researchers greatly. Developments in the research of marine microbial resources are an important basis for the study of the micro-world and deserve increasing scientific attention.

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Objective To investigate clinical presentations,laboratory examinations,magnetic resonance imaging (MRI) appearances and treatment of hypertrophic cranial pachymeningitis (HCP).Methods The clinical data of 13 patients with HCP receiving comprehensive therapy in Huashan Hospital from January 2007 to January 2013 were analyzed retrospectively.Results The onset of HCP was mostly chronic with an average duration of 26.7 months.The main clinical manifestations of the 13 patients were chronic headaches (12/13) and cranial nerve paralysis (12/13).Inflammation markers and cerebro-spinal fluid (CSF) protein levels increased in patients with HCP and gradually became normal after the treatment.The MRI demonstrated local or diffused thickened dura located in tentorium (10/13),falx cerebrum (5/13),frontal lobe (4/13),temporal lobe (7/13) and parietal lobe (4/13).The signal intensity was isointense on T1-weighted MR images and hypointense on T2-weighted MR images.Enhanced MR images showed conspicuous enhancement of the dural edges.Corticosteroid therapy improved the clinical symptoms in 12 of 13 patients.Conclusions HCP typically causes headache and paralysis of multiple cranial nerves.Enhanced MRI shows characteristic manifestations.At present corticosteroid therapy is the treatment of choice followed by immunosuppressive agent and radiotherapy.

Objective To establish HPLC determination method and impurity profile of the related substances in metronidazole.Methods A Welch Ultimate(R)XB-C1s (4.6 mm× 250 mm,5 μm)was used with a mobile phase consisting of methanol-1.36 g· L-1 solution of potassium dihydrogen phosphate (20∶ 80).The detection wavelength was 315 nm and the flow rate was 1 mL· min-1.Its related substances were determined by principal component self-contrast method.Results Good separation of metronidazole and the impurities could be achieved.Twenty batches of samples in the past six years were determined which meet quality standards.The study of impurity profiles could effectively monitor the synthetic process and the change of impurities in metronidazole.Conclusion The method is simple,quick and sensitive,which can be used to control the related substances in metronidazole.Meanwhile,the impurity profiles ensure the quality stability of metronidazole.

Objective To observe the effects of Yiqi Huoxue Tongluo Jiedu fang (YHTJF) on pneumocyte apoptosis after lung ischemia/reperfusion (I/R) injury (LIRI) in mice and to investigate whether c-Jun N-terminal protein kinase (JNK) is involved in the mechanism of apoptosis.Methods Seventy C57BL/6J male mice were randomly divided into seven groups:normal control group (C group),carboxyl methyl cellulose-Na+normal control group (CMC-Na+C group),CMC-Na+sham group (CMC-Na+S group),CMC-Na+I/R group (CMC-Na+I/R group) and CMC-Na+YHTJF-low,-middle,-high dose groups (CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups).C group did not undergo any processing;in CMC-Na+S group,only was chest opened without clipping the lung hilum;in the rest of the four groups,they all underwent opening of the chest and clipping the lung hilum for 30 minutes,then the clipping of artery was relieved and left lung reperfusion was carried out for 3 hours.After operation,the mice were sacrificed,the lung tissues were harvested.Under light and electron microscopes,the lung morphological and ultra-structural changes were observed,and the changes of index of quantitative evaluation for alveolar damage (IQA) were determined.The terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was applied to evaluate the apoptosis index (AI) of the lung tissues.The protein and mRNA expressions of JNK and glucose regulating protein 78 (GRP78) in lung tissues were detected by Western Blot and reverse transcription-polymerase chain reaction (RT-PCR);the correlations between lung AI and the expressions of mRNA and protein of JNK and GRP78,IQA were analyzed.Results Compared with CMC-Na+S group,IQA,AI and mRNA and the protein expressions of JNK and GRP78 in CMC-Na+I/R group were obviously higher [IQA:(74.00 ± 7.31)％ vs.(7.00 ± 1.23)％,AI:(64.40 ± 11.97)％ vs.(5.60 ± 1.14)％,JNK mRNA (gray value):1.143 ± 0.284 vs.0.152 ± 0.128,GRP78 mRNA (gray value):0.897 ± 0.129 vs.0.284 ± 0.044,JNK protein (A value):0.428 ± 0.074 vs.0.073 ± 0.052,GRP78 protein (A value):1.075 ± 0.145 vs.0.589 ± 0.060].Compared with CMC-Na+I/R group,the IQA,AI,protein and mRNA expressions of JNK and GRP78 in CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups were all lower,and the degree of reduction in group CMC-Na+YM was the most remarkable,greater than that in CMC-Na+YL or CMC-Na+YH group [IQA:(26.20 ± 3.35)％ vs.(34.00±5.34)％,(41.20±9.18)％,AI:(29.40±3.05)％ vs.(48.20±3.83)％,(39.20±6.14)％,JNK mRNA (gray value):0.681 ± 0.130 vs.0.804 ± 0.153,0.938 ± 0.11,GRP78 mRNA (gray value):0.450 ± 0.105 vs.0.747 ± 0.231,0.566 ± 0.115,JNK protein (A value):0.188 ± 0.049 vs.0.261 ± 0.065,0.209 ± 0.063,all P ＜ 0.01],compared with the CMC-Na+I/R group,the expression of GRP78 protein was obviously higher in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkably high was in CMC-Na+YH group (A value:1.429 ±0.226 vs.1.130±0.169,1.128 ±0.177,all P ＜ 0.01).The apoptosis of each group was mainly in the pulmonary vascular endothelial cells and alveolar epithelial cells,and brown particles were positive cells under light microscope.Under transmission electron microscope:nuclear pyknosis and margination under the nuclear membrane,cytoplasm condensed,lamellar bodies decreased and emptying increased,cell membrane microvilli decreased or disappeared,mitochondria swelling,inflammatory cells increased in alveolar septum and adhering onto the capillary walls could be seen in CMC-Na+I/R group.Compared with CMC-Na+I/R group,the lung tissue ultrastructural damage alleviated,ultrastructure of alveoli clearly seen,nuclear chromatin relatively uniform,cytoplasm increased,type Ⅱ alveolar epithelial cell surface microvilli relatively plenty,lamellar corpuscle number increased,mitochondria swelling ameliorated in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkable one was CMC-Na+YM group.AI was significantly positive correlated with the mRNA and protein expressions of JNK,GRP78 and IQA (r =0.907,0.928,0.880,0.712,0.911,all P ＜ 0.01).Conclusions YHTJF may effectively alleviate the cell apoptosis in mice LIRI,and its mechanism may be related to the inhibition of JNK pathway.

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics