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Objective:To explore the molecular mechanism of Matrine Injection in treating colorectal cancer based on network pharmacological analysis and molecular docking.Methods:Taking matrine as the object, the corresponding potential drug targets in matrine were obtained from Swiss Target Prediction database, and SuperPred database, and the database of traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP). Differential genes were obtained from gene expression omnibus (GEO), and GeneCards, OMIM, DrugBank, and CTD databases were used to collect colorectal cancer-related genes. Furthermore, core targets were screened by establishing protein-protein interaction (PPI) networks. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were performed by bioconductor of R language. Finally, the molecular docking calculation was performed to evaluate the interaction between matrine and core targets.Results:Matrine contained 63 targets. A total of 14 198 targets for colorectal cancer were obtained. The topology analysis results of the PPI network showed that 5 main targets such as myelocytomatosis proteins (MYC), interleukin-6 (IL-6), Caspase-3 (CASP3), mammalian target of rapamycin (mTOR), and amphiregulin (AR). GO enrichment analysis found that biological process (BP) mainly includes hydrogen peroxide reaction, cell reaction to hydrogen peroxide and cell response to chemical stress, etc; Cell components (CC) mainly include lipid rafts, membrane microregions and synaptic membranes, etc; Molecular functions (MF) mainly include transcriptional coregulatory factor binding, postsynaptic neurotransmitter receptor activity and core promoter sequence-specific DNA binding. KEGG pathway analysis showed that it involved chemical carcinogenesis-receptor activation, tumor necrosis factor (TNF) signaling pathway, and Toll-like receptor signaling pathway, etc. Molecular docking showed that matrine had good binding with the core target.Conclusions:Matrine acts on targets such as MYC, IL-6, CASP3, mTOR, and AR, and exerts therapeutic effects on colorectal cancer by regulating chemical carcinogenesis-receptor activation, TNF signaling pathway, Toll-like receptor signaling, etc.
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Saccharomyces boulardii is a subspecies of Saccharomyces cerevisiae and is a fungal probiotic. It can regulate the intestinal flora and enhance the barrier function of the intestinal tract. Compared with bacterial probiotics, Saccharomyces boulardii is more resistant to acid and oxidation, does not transmit genetic material with bacteria, and can be used in combination with antibiotics. Saccharomyces boulardii can function through a variety of mechanisms, and many proteases secreted by it have antitoxin effects; its own bacteria contain more polyamines, which can nourish the intestinal mucosal cells and regulate the body's metabolic balance. Besides, it can regulate multiple signal pathways to enhance intestinal immunity. Saccharomyces boulardii has been used in the treatment of ulcerative colitis (UC). The results of animal experiments and clinical studies have shown that the application of Saccharomyces boulardii can improve intestinal inflammation and enhance the therapeutic effect of mesalazine. Saccharomyces boulardii can be used as an auxiliary drug for the treatment of UC.
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Objective:To investigate the expression and clinical significance of thymidine kinase 1 (TK1), carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 19-9, CA15-3 and CA72-4 in colorectal tumors.Methods:Enzyme-linked immunosorbent assay (ELISA) and electrochemiluminescence immuno assay (ECLIA) were used to determine the serum TK1 levels and serum CEA, CA19-9, CA15-3, CA72-4 levels in 124 patients with colorectal cancer, 52 patients with colorectal precancerous lesions, 154 patients with benign colorectal lesions, and 106 health subjects. The relationship between serum TK1 and its clinicopathological characteristics in patients with colorectal cancer were analyzed. The diagnostic efficacy of TK1, CEA, CA19-9, CA15-3 and CA72-4 alone and combined detection for colorectal cancer was investigated.Results:The serum expression level of TK1 in patients with colorectal cancer was related to tumor stage, degree of differentiation, lymph node metastasis, distant metastasis and age (all P<0.05), but not related to the patient's gender ( P>0.05). The serum TK1 level decreased sequentially in colorectal cancer patients, precancerous lesions patients, benign lesions patients and healthy subjects. Colorectal cancer patients with high TK1 expression have a shorter survival time. The sensitivity, specificity and accuracy of the combined detection of TK1, CEA, CA19-9, CA15-3 and CA72-4 were 93.5%, 93.0%, and 93.1%, respectively. Conclusions:Serum TK1 is expected to become an independent marker for the diagnosis and prognosis of colorectal cancer. The combined detection of TK1, CEA, CA19-9, CA15-3 and CA72-4 has clinical significance in the diagnosis of colorectal cancer.
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The incidence of colorectal cancer is high threatening human health. About 60％～70％cases of CRC are derived from colorectal polyps, which can be treated by endoscopic electrotomy to prevent the possibility of canceration. Therefore, in the prevention and treatment of CRC, the role of screening is of great significance. CRC screening methods include the most commonly used fecal occult blood test ( FOBT ) and the more sensitive fecal immunochemical test (FIT), cost-effective fiber sigmoidoscopy and colonoscopy, CT colonoscopy (CTC), and fecal DNA testing and immature CRC hematology screening. In this paper, the CRC screening technologies were reviewed, including the principles, characteristics and the latest research progress to provide a theoretical basis for the application and development of CRC screening technology.
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Aim To investigate the effects of classic calcium antagonists verapamil(Ver),nifedipine(Nif),diltiazem(Dil)and the novel calcium antagonist N-n-butyl haloperidol iodide(F2)which was synthesized by our lab by regulating Ca2+-independent phospholipase A2(iPLA2)on hypoxia/reoxygenation(H/R)injury of cardiac microvascular endothelial cells(CMECs)and the mechanisms.Methods The CMECs were isolated from SD neonatal rats.The H/R model was established,then cells were treated with different concentrations of calcium antagonists and F2.The content of LDH in the cell supernatant was measured by colorimetric method.The levels of IL-6 and AA in cell supernatant were measured by ELISA;and late-stage apoptosis was measured by TUNEL.The mRNA and protein expression levels of iPLA2 in CMECs were examined by real time-PCR and Western blot analysis.Results Calcium antagonists except Dil decreased the generation of LDH,IL-6 and AA in a dose-dependent manner(P<0.05),and reduced the apoptosis(P<0.05).F2 and Ver decreased the mRNA and protein expression of iPLA2 in a dose-dependent manner,while there were no such effects for Nif and Dil.Conclusions Calcium antagonists except Dil have protective effects against H/R injury.F2 and Ver protect CMECs against H/R injury partly through iPLA2.
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Aim To investigate the effect of N-n-butyl haloperidol iodide(F2) on mitochondria-dependent apoptotic pathway of H9c2 cardiac myocytes during hypoxia/reoxygenation(H/R) injury.Methods The H/R models of H9c2 cardiac myocytes were established.The H9c2 cardiac myocytes were randomly divided into five groups: control group(C group), hypoxia/reoxygenation group(H/R group), F2 low concentration group(L), F2 medium concentration group(M), F2 high concentration group(H).Apoptotic rate was evaluated by flow cytometry(FCM).The levels of Cyto C, Bcl-2, Bax were observed by Western blot.Caspase-3 activity was measured with colorimetry.Results Compared with H/R group, F2 low, medium and high concentrations group could significantly decrease apoptosis rate and increase the ratio of Bcl-2 to Bax proteins and inhibit the release of Cyto C into the cytosolic fraction, and decrease caspase-3 activity.Conclusion F2 can protect H9c2 cardiac myocytes against H/R-induced injury through interfering in mitochondria-dependent pathway.
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<p><b>OBJECTIVE</b>To explore the clinical significance of vasohibin-1 expression in colorectal cancer tissues and its correlation with vascular endothelial growth factor A(VEGF-A) and microvessel density (MVD).</p><p><b>METHODS</b>Tumor tissues and paired adjacent normal tissue (distance to cancer >5 cm) from 60 colorectal cancer patients undergoing resection in the Second Hospital of Tianjin Medical University from June 2013 to November 2013 were included in this study. The protein expressions of vasohibin-1, VEGF-A and MVD were detected by immunohistochemical staining. The mRNA expressions of vasohibin-1 and VEGF-A were detected by RT-PCR. The protein expressions of vasohibin-1 and VEGF-A were observed by Western blot. Correlation among parameters was examined.</p><p><b>RESULTS</b>Vasohibin-1 expression was mainly localized in the cytoplasm of tumor cells and endothelial cells. VEGF-A expression was mainly localized in the cytoplasm and membrane of tumor cells. The expressions of vasohibin-1, VEGF-A and MVD in colorectal tumor tissues were significantly higher than those in corresponding adjacent tissues [43.3% (26/60) vs. 16.7% (10/60), 51.7%(31/60) vs. 18.3% (11/60), (39.67 ± 16.80)/mm² vs. (17.85 ± 6.43)/mm², all P<0.05]. Higher vasohibin-1 expression was significantly associated with TNM stage and metastasis (P<0.05). Vasohibin-1 expression was positively correlated with VEGF-A and MVD (r=0.378, 0.628, all P<0.05). Vasohibin-1 and VEGF-A mRNA expressions and protein expressions in colorectal cancer tissues were significantly higher than those in corresponding adjacent tissues (all P<0.05).</p><p><b>CONCLUSION</b>Vasohibin-1 expression in colorectal cancer tissues is significantly higher as compared to corresponding adjacent tissues. Vasohibin-1 expression is positively correlated to VEGF-A and MVD, and associated to TNM stage and metastasis. Positive vasohibin-1 expression indicates a poor prognosis of patients with colorectal cancer.</p>
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Humans , Cell Cycle Proteins , Colorectal Neoplasms , Microvessels , Neovascularization, Pathologic , Prognosis , Vascular Endothelial Growth Factor AABSTRACT
Aim To establish a novel,highly sensitive,rapid and cost-effective HPLC method to simultaneously determine tri-cyclic antidepressant clomipramine and four benzodiazepines of diazepam,alprazolam,clonazepam,oxazepam in human plasma pretreated by solid phase extraction.Methods The assay was achieved by using C8 column (4.6 mm ×1 50 mm,5 μm)kept at 45 ℃,mobile phase 73.2:26.8 V/V (50 mmol·L -1 ,pH 3.0 phosphate buffer:acetonitrile)with flow rate of 1 .2 ml· min -1 ,and UV detection was set at λ220 nm.Solid phase ex-traction was performed on C1 cartridges.Results The calibra-tion curve was demonstrated to be linear (r >0.9994)in the ranges of 5 ~200 μg·L -1 for alprazolam and clonazepam,1 0 ~500 μg·L -1 for diazepam,20 ~500 μg· L -1 for clomipra-mine,and 7.5 ~2000 μg·L -1 for oxazepam;the limit of detec-tion (LOD)was 1 .5,1 .4,3.0,5.5 and 2.2 μg·L -1 for al-prazolam,clonazepam,diazepam,clomipramine and oxazepam respectively.Intra-day and inter-day precision revealed a coeffi-cient of variation of 2.2% ~1 2.6% and 2.1 % ~1 3.2%,re-spectively.Extraction yield ranged from 81 .1 % ~1 00.1 % for all analytes.Conclusion The developed method is accurate, reproducible,convenient,and suitable for routine therapeutic drug monitoring of clomipramine and the four benzodiazepines.
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<p><b>OBJECTIVE</b>To compare the clinical effect of graciloplasty using two different gracilis encircled loops and to research the better method for the treatment of fecal incontinence after anoplasty for imperforate anus.</p><p><b>METHODS</b>Clinical data of 38 incontinence patients with congenital anal malformation undergoing graciloplasty in our hospitals from January 2009 to January 2012 were retrospectively analyzed. Twenty patients received the modified surgery in which the gracilis muscle was transposed anticlockwise into a circum-anal tunnel with a U-loop and its tendon secured to the ipsilateral pectineal ligament. Eighteen patients received the traditional surgery in which the gracilis muscle was wrapped clockwise around the anus with a γ-loop and its tendon secured to the contralateral periosteum of ischial tuberosity or pectineal ligament. All the patients were evaluated via Wexner score and anal manometry before surgery and 2 weeks, 6 months, 1 year and 2 years after surgery. In addition, it was assessed whether the patients had difficulty defecating while squatting after surgery.</p><p><b>RESULTS</b>Generalized estimating equations showed that the Wexner score in two groups gradually decreased after surgery (P=0.000), but there was no significant difference between two groups (P=0.554). At 2 weeks, 6 months, 1 year and 2 years after surgery, there were respectively 4 cases (20.0%), 3 cases (15.0%), 2 cases (10.0%), 1 case (5.0%), who showed squatting difficult defecation in the U-loop group, and 10 cases (55.6%), 12 cases (66.7%), 10 cases (55.6%), 8 cases (44.4%) in the γ-loop group. The probability of squatting difficult defecation in U-loop group was significantly lower compared to γ-loop group (P=0.002), but the probability of squatting difficult defecation in two groups did not obviously change with time (P=0.057). Repeated measures ANOVA showed that anal resting pressure, anal maximal squeeze pressure, duration of anal squeeze, and rectal maximum tolerable volume in two groups were gradually improved after surgery (all P<0.01), but there were no significant differences between two groups (all P>0.05).</p><p><b>CONCLUSIONS</b>Graciloplasty with different gracilis loops can improve anal function. However, U-loop can better improve difficult defecating while squatting, and is worthwhile for spreading in clinical practice.</p>
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Humans , Anal Canal , Anus, Imperforate , Digestive System Surgical Procedures , Electric Stimulation Therapy , Fecal Incontinence , PressureABSTRACT
Aim To test the skin healing and repairing efficacy and the mechanism of Hibiscus rosa-sinensis L bud extract by using the animal models. Methods KM mice were randomly divided into three groups:the model group, the positive control group, and the n-bu-tyl alcohol extract ( HrBN) group. Using the boils and carbuncles model, the healing condition of all the animals were observed. KM mice were kept in the SPF condition room and divided into five groups: the model group, the positive control group, and the low, middle, high dose groups. Using the full-thickness loss model, the repairing results of all the mice were ob-served. Through the antimicrobial test, the results of MIC and inhibition zone were obtained. The carbon clearance test was used to collect the blood at the time 5min and 15min, and get the liver and spleen, and the results of K andαwere obtained. Results In vivo ex-periments showed there was significant difference be-tween groups;the HrBN extract had the outstanding ef-ficacy in healing and repairing skin boils and full-thickness loss models. It had higher recovery rate than other ethanol extract, such as ethyl acetate extract and chloroform extract. In vitro experiments showed that the HrBN extract, ethyl acetate extract ( HrBE) ,AB-8 macroporous resin 30% alcohol part and 60% alcohol part had obvious antimicrobial efficacy. The carbon clearance test showed HrBN had a good effect in im-proving immune function, and it can increase the K and α. Conclusion HrBN in animal models exerts good skin healing and repairing efficacy, which might be related to its antibacterial activity and immunologic enhancement function.
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Angiogenesis is regulated by the local balance between angiogenesis stimulators and inhibitors. Vasohibin family as a novel regulator of angiogenesis involves in the process of various pathophysiology. Vasohibin-1 is the only factor that regulates angiogenesis by means of negative feedback as the mechanism of action. It is expressed in ECs to terminate an-giogenesis. Subsequently one gene homologous to vasohibin-1 was found and designated as vasohibin-2, which is expressed in infiltrating MNCs or cancer cells to stimulate angiogenesis. Furthermore, one binding protein to vasohibin-1 and vasohib-in-2 has been discovered and renamed as small vasohibin-binding protein (SVBP). The analysis of the function of SVBP has revealed that SVBP binds to vasohibin-1 within cells, makes a heterodimer with vasohibin-1 and facilitates the secretion of vasohibin-1. This article reviews the construction features and biological activities of vasohibin family members and their ef-fects of inhibiting pathology angiogenesis.
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Aim To construct eukaryotic expressing plasmid of hi FGF2 ( high molecular weight isoform fi-broblast growth factor-2,hi FGF2) gene and to investi-gate its effect on apoptosis after its overexpression in HEK293 cells. Methods The DNA template primer was designed and synthesized. The pDsRed1-N1 plas-mids were digested by the restriction enzymes of Nhel and Hind III. The hi FGF2 was ligated with linearized pDsRed1-N1 by T4 DNA Ligase. The recombinant plasmid was identified by endonuclease digestion and sequenced. The recombinant hi FGF2 plasmid was transient transfected into HEK293 cells by Lipofectami-neTM 2000 Reagent. The transfection efficiency was de-tected by fluorescence inversion microscope. The cell apoptosis was detected by Annexin V-FITC/PI apopto-sis detection kit with flow cytometry analysis. Results The pDsRed1-N1 eukaryotic expression vector was consistent with the design. The recombinant hi FGF2 plasmid was transfected in HEK293 cells. The trans-fection rate was more than 70%. The FITC/PI dyeing rate in hi-FGF2 over-expression HEK297 cells was a-bout ( 29. 12 ± 2. 81 )%. Conclusions pDsRed1-N1 eukaryotic expression vector is successfully constructed and transfected into HEK293 cells. Over-expression of hi FGF2 induces cell apoptosis.
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Objective To investigate possible disturbance of Th1/Th2 immunoregulation of blood serum and bladder mucosa in patients with interstitial cystitis(IC). Methods Blood serum and bladder mucosal specimens were collected from 16 female patients with IC and 16 female normal controis.The age of IC patients was 51.3±10.2 years.The age of normal controls was 53.1±9.6years.The expressions of Th1(IFN-γ,IL-2)and Th2 cytokines(IL-4,IL-10)were determined by enIFN-γ and IL-2 in IC blood serum were(4.57±2.92),(17.52±7.52)pg/ml and in normal blood serum were(4.11±2.27),(20.99±5.09)pg/ml.Such two factors had no significant differences in these two groups.The expressions of IL-4 and IL-10 in IC blood serum were(0.34±0.22),(4.37±2.34)pg/ml and in normal blood serum were(O.14±O.07),(1.18±0.61)pg/ml.Such two factors had sighad no significant differences with normal bladder tissues(IFN-γ:χ2＝1.900,IL-2:χ2＝0.514).The expressions of IL-4 and IL-10 in IC bladder tissues had significant differences with normal bladder tiswere(0.16±0.11)and(2.42±1.27)pg/ml but in high PUF group were(0.53±0.12)and(6.31±1.21)pg/ml.The expressions of these two factors had significant correlations with the degree of PUF,which intensified in the IC of high PUF. Conclusion The expression of Th2 type eytokines is predominant in IC blood plasm and bladder tissues,the expression of Th2 type cytokines in IC blood plasm and bladder tissues has positive correlation with the degree of PUF.
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AIM To study the effect of quaternary ammonium salt derivative of haloperidol (F 2) on L type calcium current ( I Ca ) in rat ventricular myocytes. METHODS Single ventricular cell of rat was obtained by enzymatic dissociation method. The currents were recorded with the whole cell configuration of the patch clamp technique. RESULTS F 2(1 ?mol?L -1 ) decreased I Ca from ( 1 775 2 ?360 4) pA to (464?129 1) pA ( n =8, P
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AIM To observe the effect of quateranary ammonium salt derivative (F2) of haloperidol on calcium level in vascular smooth muscle cells (VSMCs) isolated from thoracic aortas of rat. METHODS VSMCs were loaded with Fluo-3-AM, a calcium sensitive fluorescent dye, and ［Ca2+］i was determined by the use of laser scanning confocal microscope (LCSM). RESULTS In Ca2+ (1.26 mmol*L-1) bath solution, intracellular calcium fluorescent intensity (FI) in VSMCs was increased when application with 30 mmol*L-1 KCl, and then was inhibited after addition with F2. The FI of the ASMCs with different concentrations of F2 (0.1,1,10,100 μmmol*L-1)within three minutes were 64%±9%(n=16, P<0.05),16%±5%(n=20, P<0.01),0.6%±0.8%(n=20, P<0.01),0.1%±0.3%(n=15, P<0.01), respectively. CONCLUSION F2 could decrease the intracellular Ca2+ level in the VSMCs by blocking the voltage-dependent calcium channel.
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Antisense oligonucleotides is appplicated abroadly in cardiovascular disease.It aims at the target genes involved in molecule mechanisms of disease pathogenesis.This technology reveals good foreground as a kind of clinical drug because of its researchful and therapeutic function in hypertension、coronary disease,complications of heart transplantation and failure.
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Electromagnetic blood flowmeter (EBF) and polygraph system were used for measuring the coronary blood flow (CBF), left ventricular systolic pressure (LVSP),mean aortic pressure (MAP ), heart rate respectively in anesthetized dogs. In group 1 (6 dogs ), after opening the chest,the left coronary artery was surrounded by a fitting flow prob of EBF and ploygraph system was connected with dogs. morphine (2mg. kg-1 ),morphine plus naloxone were intravenously injected into dogs respectively. It showed that CBF was increased 56. 8% with morphine, while the LVSP and MAP decreased, HR not changed. The effect mentioned above were partly antagonized by naloxone (0. 4mg/kg). In group 2(3 dogs) using same methods, HR.CBF. LVSP and MAP were decreased with high dose fentanyl (100ug/kg ). The results suggested that in clinic the use of high dose fentanyl should be careful during anesthesia for the patients with cardiopathy. However, the effect of morphine in increasing CBF may be beneficial for the patients with myocardial ischemia.
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AIM To observe the effect of quateranary ammonium salt derivative (F 2) of haloperidol on calcium level in vascular smooth muscle cells (VSMCs) isolated from thoracic aortas of rat. METHODS VSMCs were loaded with Fluo 3 AM, a calcium sensitive fluorescent dye, and [Ca 2+ ] i was determined by the use of laser scanning confocal microscope (LCSM). RESULTS In Ca 2+ (1 26 mmol?L -1 ) bath solution, intracellular calcium fluorescent intensity (FI) in VSMCs was increased when application with 30 mmol?L -1 KCl, and then was inhibited after addition with F 2. The FI of the ASMCs with different concentrations of F 2 (0 1,1,10,100 ?mmol?L -1 )within three minutes were 64%?9%( n =16, P
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AIM To study the effects of quateranary ammonium salt derivative (F 2) of haloperidol on ischemia and reperfusion injury in rat hearts. METHODS Ischemia and reperfusion injury in rat hearts was induced by occluding the left anterior descending coronary artery for 30 min and restoring blood reperfusion for 30 min. F 2 (1, 2, 4 mg?kg -1 , respectively) was intravenously injected before heart ischemia. Plasma creatine kinase (CK), creatine kinase isoenzyme MB(CK MB), lactate dehydrogenase(LDH),? Hydroxybutyrate dehydrogenase (HBDH), grutamic oxalacetic transaminase(GOT), superoxide dismutase (SOD) activity and malondiadehyde (MDA) contents were measured. The pathologic changes of ischemia and reperfusion myocardium were observed on the transmission electron microscopy. RESULTS F 2 reduced the release of CK,CK MB LDH,HBDH,GOT from I/R rat hearts, increased the activity of SOD and decreased the MDA contents. In F 2 (1mg?kg -1 ) group, the serum CK MB LDH HBDH concentration was lowered significantly (vs I/R group P
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AIM To investigate the effect of N n butyl haloperidol iodide (F 2) on potassium currents in enzymatically isolated vascular smooth muscle cells (VSMC) from thoracic aortas and the effect of F 2 on aortic rings of rat. METHODS The whole cell patch clamp technique and the contraction of rats thoracic aortic rings were used in experiments. RESULTS The outward currents were observed when holding potential was -40 mV and the cell was depolarized from -30 mV to +100 mV (in 10 mV increase) for 400 ms. At the point of the test potential of +70 mV, solutions of F 2 (0 1,1, 5 ?mol?L -1 ) were added into bath (external) solution, which led to the increase of the outward currents from (229?28)pA,(226?57)pA and(228?42) pA to (354?29) pA ( n =6, P