[Mechanism of Euodiae Fructus stir-fried with water decoction of Coptidis Rhizoma in treatment of chronic colitis based on UPLC-Q-TOF-MS, network pharmacology and experimental verification].

To elucidate the mechanism of Euodiae Fructus stir-fried with water decoction of Coptidis Rhizoma in the treatment of chronic colitis, this study employed ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS), network pharmacology, and experimental verification to predict the involved targets and signaling pathways. The chronic colitis mouse model was constructed to verify the core targets. A total of 48 compounds in the herbal medicine were identified by UPLC-Q-TOF-MS. SwissTargetPrediction was used to screen the potential active components and drug targets. GeneCards, OMIM, PharmGKB, and TDD were used to search for the disease targets. A total of 31 active ingredients, 453 targets of the herbal medicine, and 3 960 targets of chronic colitis were obtained. The common targets shared by the herbal medicine and chronic colitis were introduced into STRING to construct the protein-protein interaction(PPI) network, and CytoNCA plug-in was used to screen the key targets. A total of 90 key targets were obtained, and the key active components included isorhamnetin, quercetin, limonin, and oxyberberine. GO annotation and KEGG pathway enrichment for the key targets were carried out via DAVID. The targets were mainly involved in the positive regulation of protein phosphorylation, positive regulation of nitric oxide biosynthetic process, and negative regulation of apoptotic process. The medicine may treat chronic colitis through PI3 K-Akt, VEGF, HIF-1, and TNF signaling pathways. A mouse model of chronic colitis was established and then treated with Euodiae Fructus stir-fried with the water decoction of Coptidis Rhizoma. The experimental results demonstrated that the medicine can alleviate the pathological damage of colon, significantly reduce the levels of IL-1ß, IL-6, and TNF-α, inhibit the activation of PI3 K/Akt pathway, and down-regulate the expression of VEGFA in the treatment of chronic colitis.

Modified membrane fixation technique in a severe continuous horizontal bone defect: A case report.

BACKGROUND: Continuous severe horizontal bone defect is common in the aesthetic maxillary anterior area, and presents a major challenge in implant dentistry and requires predictable bone augmentation to increase the width of the alveolar bone. CASE SUMMARY: A 24-year-old man, with a history of well-controlled IgA nephropathy, presented to the Dentistry Department of our hospital complaining of missing his right maxillary anterior teeth 1 mo ago. Severe horizontal alveolar bone defects at sites of teeth 12, 13 and 14 were diagnosed. A modified guided bone regeneration surgical approach stabilizing the absorbable collagen membrane and particulate graft materials by periosteal diagonal mattress suture (PDMS) combined with four corner pins was used for this severe continuous horizontal bone defect. The outcome revealed that the newly formed alveolar ridge dimension increased from 0.72 mm to 11.55 mm horizontally 10 mo postoperatively, with no adverse events. The implant surgery was successfully performed. CONCLUSION: This case highlights that PDMS combined with four corner pins is feasible to maintain the space and stabilize the graft and membranes in severe continuous horizontal bone defect.

Circular RNA circVAPA promotes chemotherapy drug resistance in gastric cancer progression by regulating miR-125b-5p/STAT3 axis.

BACKGROUND: Gastric cancer (GC) is a prevalent malignancy, leading to a high incidence of cancer-associated death. Cisplatin (DDP)-based chemotherapy is the principal therapy for clinical GC treatment, but DDP resistance is a severe clinical challenge and the mechanism remains poorly understood. Circular RNAs (circRNAs) have been identified to play crucial roles in modulating the chemoresistance of gastric cancer cells. AIM: To explore the effect of circVAPA on chemotherapy resistance during GC progression. METHODS: The effect of circVAPA on GC progression and chemotherapy resistance was analyzed by MTT assay, colony formation assay, Transwell assay, wound healing assay, and flow cytometry analysis in GC cells and DDP resistant GC cell lines, and tumorigenicity analysis in nude mice in vivo. The mechanism was investigated by luciferase reporter assay, quantitative real-time PCR, and Western blot analysis. RESULTS: CircVAPA expression was up-regulated in clinical GC tissues compared with normal samples. CircVAPA depletion inhibited proliferation, migration, and invasion and increased apoptosis of GC cells. The expression of circVAPA, STAT3, and STAT3 downstream genes was elevated in DDP resistant SGC7901/DDP cell lines. CircVAPA knockdown attenuated the DDP resistance of GC cells. Mechanically, circVAPA was able to sponge miR-125b-5p, and miR-125b-5p could target STAT3 in the GC cells. MiR-125b-5p inhibitor reversed circVAPA depletion-enhanced inhibitory effect of DDP on GC cells, and STAT3 knockdown blocked circVAPA overexpression-induced proliferation of DDP-treated SGC7901/DDP cells. The depletion of STAT3 and miR-125b-5p inhibitor reversed circVAPA depletion-induced GC cell apoptosis. Functionally, circVAPA contributed to the tumor growth of SGC7901/DDP cells in vivo. CONCLUSION: CircVAPA promotes chemotherapy resistance and malignant progression in GC by miR-125b-5p/STAT3 signaling. Our findings present novel insights into the mechanism by which circVAPA regulates chemotherapy resistance of GC cells. CircVAPA and miR-125b-5p may be considered as the potential targets for GC therapy.

[Determination of trace lead in water and milk tea powder samples with organic coprecipitation-flame atomic absorption spectrometric].

A method was proposed for the determination of trace lead with flame atomic absorption spectrometry after preconcentration of lead by rapid coprecipitation technique with PAR-Fe (III) at pH 6.0. The analytical parameters including pH, amount of iron (III), amount of reagent, the standing time of the precipitate, etc., were examined. The detection limits (DL) were found to be 18.7 microg x L(-1) for Pb (II). In analysis of lake water and the milk tea powder samples, RSD's and the standard addition recovery of this method were in the ranges of 1.03%-2.24% and 94.2%-98.3% respectively. The effect of matrix can be overcome by the method and the results are satisfyiog. The method shows good application prospect in the determination of trace lead owing to its rapidness and reproducibility.

Downregulation of miR-497 promotes tumor growth and angiogenesis by targeting HDGF in non-small cell lung cancer.

MicroRNAs (miRNAs) play important roles in the development of various cancers. MiRNA-497 functions as a tumor-suppressor that is downregulated in several malignancies; however, its role in non-small cell lung cancer (NSCLC) has not been examined in detail. Here, we showed that miR-497 is downregulated in NSCLC tumors and cell lines and its ectopic expression significantly inhibits cell proliferation and colony formation. Integrated analysis identified HDGF as a downstream target of miR-497, and the downregulation of HDGF by miR-497 overexpression confirmed their association. Rescue experiments showed that the inhibitory effect of miR-497 on cell proliferation and colony formation is predominantly mediated by the modulation of HDGF levels. Furthermore, tumor samples from NSCLC patients showed an inverse relationship between miR-497 and HDGF levels, and ectopic expression of miR-497 significantly inhibited tumor growth and angiogenesis in a SCID mouse xenograft model. Our results suggest that miR-497 may serve as a biomarker in NSCLC, and the modulation of its activity may represent a novel therapeutic strategy for the treatment of NSCLC patients.

Expression of homeobox gene MSX-2 during cranial suture fusion of SD rats / 上海第二医科大学学报

Objective To investigate the expression of homeobox gene MSX-2 during cranial suture fusion of SD rats and discuss its significance. Methods SD rats aged 1, 2, 5, 8, 12, 15, 18, 22, 30 and 45 days were selected, and immunohistochemistry and Real-time PCR were employed to localize and quantify the expression of MSX-2 in different regions of cranial sutures. Results MSX-2 expressed in calvarial suture tissues including the extreme ends of the osteogenic fronts and the underlying dura mater. The expression of MSX-2 was low in posterior frontal suture (PF) and sagittal suture (SAG) from postnatal day 1 to day 8 before the initiation of suture fusion, while it was higher in PF than in SAG from postnatal day 12 to day 22 after the initiation of PF suture fusion. The expression of MSX-2 significantly declined in PF and was moderately higher than that in SAG from postnatal day 30 to day 45 after the initiation of suture fusion. Conclusion There is different expression of MSX-2 in PF and SAG during different suture fusion periods, which suggests the expression of MSX-2 may participate in the regulation of cranial bone development and the fusion of cranial sutures.

Construction and functional identification of eukaryotic expression vector carrying Sprague-Dawley rat MSX-2 gene / 中华整形外科杂志

<p><b>OBJECTIVE</b>To construct a high effective eukaryotic expressing plasmid PcDNA 3.1-MSX-2 encoding Sprague-Dawley rat MSX-2 gene for the further study of MSX-2 gene function.</p><p><b>METHODS</b>The full length SD rat MSX-2 gene was amplified by PCR, and the full length DNA was inserted in the PMD1 8-T vector. It was isolated by restriction enzyme digest with BamHI and Xhol, then ligated into the cloning site of the PcDNA3.1 expression plasmid. The positive recombinant was identified by PCR analysis, restriction endonudease analysis and sequence analysis. Expression of RNA and protein was detected by RT-PCR and Western blot analysis in PcDNA3.1-MSX-2 transfected HEK293 cells.</p><p><b>RESULTS</b>Sequence analysis and restriction endonudease analysis of PcDNA3.1-MSX-2 demonstrated that the position and size of MSX-2 cDNA insertion were consistent with the design. RT-PCR and Western blot analysis showed specific expression of mRNA and protein of MSX-2 in the transfected HEK293 cells.</p><p><b>CONCLUSIONS</b>The high effective eukaryotic expression plasmid PcDNA3.1-MSX-2 encoding Sprague-Dawley Rat MSX-2 gene which is related to craniofacial development can be successfully reconstructed. It may serve as the basis for the further study of MSX-2 gene function.</p>

Evaluation of liquid based cytology test (LCT) in avoiding medical tangles / 法医学杂志

OBJECTIVE@#To evaluate of liquid based cytology test (LCT) in avoiding medical tangles.@*METHODS@#One thousand five hundred five thirty one cases, which were collected from out-patients of precancerous lesions of uterine cervix, were randomly divided into three groups based on different smear preparation: LCT method was used in two groups (one with ThinPrep kit and one with ArtoBrain kit), conventional Papauicolaou smear (PS) was used in one group. All cases of abnormal cervical smears were identified by cytologic test underwent colposcopic examination and colopscopically multiple biopsy. Results of test were analyzed by software SPSS 11.0.@*RESULTS@#Significant diference were found between LCT method and PS method compared by index of satifacation, sensitivity, specificity, accuracy, false negative rate and erroneous diagnosis rate (P < 0.05, but no difference were found between two LCT groups (ThinPrep kit and ArtoBrain kit).@*CONCLUSION@#LCT method can improve diagnostic level of precancerous lesions of uterine cervix either tested by ThinPrep kit or ArtoBrain kit, so have the powerfull value to avoid medical tangles.

[Study on the antibacterial mechanisms of copper-bearing montmorillonite].

Vibrio parahaemolyticus ATCC 27519 was chosen as indicator of aquacultural pathogenic bacteria to determine the antibacterial activity and mechanism of copper-bearing montmorillonite (Cu-MMT) in vitro. The results indicated that montmorillonite (MMT) had no antibacterial activity. The minimum inhibitory concentration (MIC) and bactericidal concentration (MBC) of Cu-MMT on Vibrio parahaemolyticus were 75 and 300 mg/L, respectively. The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) of bacteria were examined and the results showed treatment with Cu-MMT could lead to significant release of intracellular enzymes from the tested bacteria suggesting that the permeability of the cell membrane increased and bacteria suffered injury. Three typical inhibitors (malonic acid, iodine acetic acid and phosphate sodium) were used to further study the inhibitory pathways of respiratory metabolism. Cu-MMT effectively inhibited respiratory metabolism of Vibrio parahaemolyticus, with the respiratory inhibition percent (I(R)) of 31.8%. The respiratory superposing inhibition percent after addition of phosphate sodium, iodine acetic acid and malonic acid was 48.6%, 27.8% and 17.5%, respectively. These results indicated that the effect of malonic acid on superposing inhibition percent of Cu-MMT for bacteria is the lowest; thus, the synergic action between Cu-MMT and malonic acid is the weakest, indicating that they inhibited the same pathway of respiratory metabolism, i.e. the TCA pathway, which is the most important pathway of carbohydrate metabolism. The atomic force microscope image of Vibrio parahaemolyticus exposed to Cu-MMT showed that Cu-MMT could rupture the bacterial cell membrane.

[Effects of granulocyte colony-stimulating factor on peripheral endothelial progenitor cells in cholesterol-fed rabbits].

OBJECTIVE: To investigate the effects of granulocyte colony-stimulating factor (G-CSF) on peripheral endothelial progenitor cells (EPC) and atherosclerosis (AS) in cholesterol-fed rabbits. METHODS: Male New Zealand white rabbits were randomly divided into control group, G group (Recombinant Human Granulocyte Colony Stimulating Factor Injection rhG-CSF 50 microg/d), AS group (high cholesterol diet) and G + AS group (rhG-CSF 50 microg/d plus high cholesterol diet, n = 8 per group). Peripheral blood was collected at baseline and at 1, 4, 8 and 12 weeks, total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After being cultured for 7 days, EPCs were characterized as adherent cells double positive for DiLDL uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. After being cultured for 3 days, the number of EPC (PE-CD34/FITC-CD133 double-stained positive cells) was quantified by flow cytometric analysis. Serum NO was measured and aortic plaque area analyzed at 12 weeks. RESULTS: EPC number was low in control and AS groups and EPC number was significantly increased ( approximately 13-fold, P < 0.001) compared to baseline at 1 week in G and G + AS groups and remained at this level throughout the study period in G group while decreased gradually in G + AS group and returned to baseline level at 12 weeks. Aortic atherosclerotic plaque was visible in both AS and G + AS groups, however, the aortic atherosclerotic plaque area was smaller in G + AS group than that of in As group (59.8 mm(2) +/- 26.9 mm(2) vs. 251.5 mm(2) +/- 83.4 mm(2), P < 0.01). Serum NO was similar between AS and G + AS groups and significantly higher than that in control and G groups. CONCLUSION: CSF could attenuate atherosclerosis in cholesterol-fed rabbits by increasing circulating EPC.

[Time course of G-CSF, estrogen and various doses of atorvastatin on endothelial progenitor cells mobilization].

OBJECTIVE: To evaluate the time course of granulocyte-colony-stimulating-factor (G-CSF), estrogen and various doses of atorvastatin on endothelial progenitor cells (EPCs) mobilization. METHOD: A total of 48 male New Zealand White rabbits were treated with placebo, estrogen (0.25 mg.k(-1).d(-1)), Atorvastatin (2.5, 5, or 10 mg) and G-CSF (50 microg/rabbit/d), respectively. Peripheral EPCs number was surveyed weekly for 4 weeks by FACS analysis (double-positive for PE-CD34/FITC-CD133) and under fluorescent microscope (double-positive for FITC-UEA-1/Dil-acLDL). Serum nitric oxide (NO) and lipids were also measured at the third week. RESULTS: Peripheral EPCs was significantly increased in G-CSF treated animals and remained constant for 4 weeks compared to placebo treated animals. Atorvastatin increased peripheral EPCs dose-dependently from 2.5 to 5 mg and peaked at the third week while peripheral EPCs number was not affected by 10 mg.k(-1).d(-1) atorvastatin during the first 3 weeks and was significantly higher only in the fourth week compared to placebo group. Estrogen also significantly increased peripheral EPCs at the third and fourth week compared to placebo group. At the third week, serum NO was similar in G-CSF group, significantly higher in atorvastatin 5 mg.k(-1).d(-1) and estrogen groups while significantly lower in atorvastatin 10 mg.k(-1).d(-1) group compared to placebo group. Serum lipids were similar among various groups. CONCLUSION: Atorvastatin, estrogen and G-CSF could mobilize EPCs. The mobilization efficacy is as follows: G-CSF > atorvastatin 5 mg.k(-1).d(-1) > estrogen > atorvastatin 2.5 mg.k(-1).d(-1) > atorvastatin 10 mg.k(-1).d(-1). NO might partly contribute to the mobilizing effect of estrogen and atorvastatin.

Expression of CDK4 and cyclin D1 proteins in gastric carcinoma / 上海第二医科大学学报

Objective To investigate the expression and significance of CDK4 and cyclin D1 in gastric carcinoma,and to discuss their correlation with WHO histological classification,TNM stages and lymph node metastasis.Methods Forty paraffin wax specimens from patients with gastric carcinoma were collected and expression of CDK4 and cyclin D1 proteins were detected by immunohistochemistry.Results Immunohistochemistry demonstrated that the positive rates for CDK4 and cyclin D1 protein were 65% and 55%,respectively.There was positive correlation between CDK4 and cyclin D1 proteins(P0.05).Conclusion CDK4 and cyclin D1 proteins are frequently overexpressed in gastric carcinoma and there is a positive relationship between the two proteins,which may be involved in the development of gastric carcinoma.