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Objective To investigate the effect of Echinococcus granulosus cyst fluid(EgCF) on the cytoskeletal rearrangement and phagocytosis and the migration of macrophages induced by lipopolysaccharide(LPS). Methods Peritoneal macrophages of C57BL/6 mice were isolated and cultured in vitro, and divided into control group and LPS group and LPS combined with EgCF group. After 48 hours of treatment, filamentous actin (F-actin) changes were observed with rhodamine-labelled phalloidin staining and fluorescence microscopy; TranswellTM chamber was used to test cell migration ability and flow cytometry to test cell phagocytosis. After 1 hour of treatment, PI3K and AKT, phosphorylated AKT (p-AKT), Rac1, guanosine triphospho-Rac1 (GTP-Rac1), WASP and Arp2 protein expressions were detected with Western blot analysis. Results Compared with the control group, after LPS stimulation, macrophages were deformed significantly; pseudopodia increased; actin cytoskeleton increased and was more distributed in pseudopodia; the ability of migration and phagocytosis were significantly improved, and the expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 proteins significantly increased. EgCF treatment caused cell shrinkage and disappearance of pseudopodia protrusions of LPS-activated cells, and led to the reduced phagocytic and migratory of cells; the protein expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 decreased significantly compared with the LPS group. Conclusion LPS induces the migration and enhances phagocytosis of macrophages while EgCF inhibits these effects, which is related to actin cytoskeleton rearrangement.
Subject(s)
Mice , Animals , Lipopolysaccharides/pharmacology , Echinococcus granulosus/metabolism , Proto-Oncogene Proteins c-akt , Cyst Fluid/metabolism , Mice, Inbred C57BL , Macrophages/metabolism , Phagocytosis , Actins/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Guanosine Triphosphate/pharmacologyABSTRACT
Objective:To investigate the therapeutic effects of a small interfering RNA (siRNA) targeting hypoxia inducible factor-1α (HIF-1α) on collagen-induced arthritis (CIA) in mice and to analyze the possible mechanism at the macrophage level.Methods:DBA/1 mice were injected with bovine type Ⅱ collagen to establish the CIA model. Then, they were injected with HIF-1α-siRNA adenovirus or negative control adenovirus through tail vein once a week for four weeks. This study included four groups: control group, CIA model group, negative control group and HIF-1α-siRNA group. Mouse bone marrow-derived macrophages (BMDMs) were isolated and cultured. The relative expression of CD206 and arginine (Arg) at mRNA level in mouse BMDMs was detected by RT-PCR. The proportions of F4/80 + CD16/32 + M1 and F4/80 + CD206 + M2 macrophages in spleen and thymus were detected by flow cytometry. Pathological changes in the ankle joint of mice were observed using HE staining. Immunohistochemistry was used to detect the expression of macrophages and the subsets in mouse synovial tissues. Results:(1) Compared with the control group, the CIA model group showed decreased expression of CD206 at mRNA level in BMDMs, but increased expression of Arg at mRNA level ( P<0.01). HIF-1α-siRNA increased the expression of CD206 at mRNA level ( P<0.05) and reduced the expression of Arg at mRNA level in BMDMs of mice with CIA ( P<0.01). (2) Compared with the control group, the mice in the CIA model group had increased proportions of F4/80 + CD16/32 + M1 macrophages in splenocytes and thymocytes ( P<0.05), but decreased proportions of F4/80 + CD206 + M2 macrophages in thymocytes ( P<0.05). HIF-1α-siRNA could down-regulate the proportions of F4/80 + CD16/32 + M1 macrophages in splenocytes and thymocytes ( P<0.05), and up-regulate the proportions of F4/80 + CD206 + M2 macrophages in thymocytes of CIA mice ( P<0.01). (3) CIA mice had synovial hyperplasia and macrophages infiltration, especially M1 macrophages, in the ankle joint. HIF-1α-siRNA could alleviate the synovial hyperplasia and macrophage infiltration. Conclusions:HIF-1α-siRNA could alleviate macrophage infiltration and synovial hyperplasia in CIA mice through reducing the proportions of M1 macrophages in thymocytes, BMDMs and synovial tissues and increasing the proportion of M2 macrophages, suggesting that HIF-1α-siRNA could treat CIA mice by regulating the differentiation of M1 and M2 macrophages.
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Objective To evaluate the role of polysaccharide from Phellinus igniarius (PPI) in the improvement of oxidative stress, hepatic granuloma and hepatic fibrosis in Schistosoma japonicum-iniected in mice. Methods The mouse model of schistosomiasis was established by S. japonicum cercariae infection via the abdomen. Balb/c mice were randomly assigned into 5 groups, including the healthy control group (Group A), infection control group (Group B), PPI treatment group (Group C), praziquantel treatment group (Group D) and PPI-praziquantel combination group (Group E), of 10 mice in each group. Each mouse in groups B, C, D and E was infected with (30 ± 2) S. japonicum cercariae. Then, mice in groups D and E were given praziquantel by gavage at a dose of 500 mg/kg for successive two days on day 42 post-infection, while mice in groups C and E were given PPI by gavage at a dose of 400 mg/kg for successive 30 days on day 42 post-infection. Histopathological changes of hepatic tissues were observed using hematoxylin-eosin (HE) staining, and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN) were determined, while the activities of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), glutathione reductase (GSH-R) and glutathione (GSH) were detected in Mouse liver homogenates. The expression of transforming growth factor-beta (TGF-β) and alpha-smooth muscle actin (α-SMA) was quantified in hepatic tissues using immunohistochemistry, and the Nrf2 and Gsta4 gene expression was quantified using quantitative real-time PCR (qPCR) assay. Results Untreated mice presented typical pathological changes of schistosomal hepatic disorders, while PPI treatment effectively alleviated hepatic egg granulomas and collagen deposition. S. japonicum infection resulted in aggravation of hepatic lipid peroxidation, induction of oxidative stress, elevated serum MDA level and a reduction in the activity of GSH and antioxidant enzymes activities in mice. As compared to infected but untreated mice, PPI treatment suppressed hepatic lipid peroxidation, increased the GSH activity and restored the activity of antioxidant enzymes. In addition, PPI treatment inhibited the TGF-β signaling pathway and up-regulated the Nrf2 and Gsta4 gene expression. Conclusions PPI plays a critical role in the treatment of schistosomiasis-induced hepatic fibrosis. It may improve oxidative stress damages through up-regulating Nrf2 and Gsta4 gene expression, thereby suppressing the development of hepatic egg granulomas and hepatic fibrosis.
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Objective To evaluate the role of polysaccharide from Phellinus igniarius (PPI) in the improvement of oxidative stress, hepatic granuloma and hepatic fibrosis in Schistosoma japonicum-iniected in mice. Methods The mouse model of schistosomiasis was established by S. japonicum cercariae infection via the abdomen. Balb/c mice were randomly assigned into 5 groups, including the healthy control group (Group A), infection control group (Group B), PPI treatment group (Group C), praziquantel treatment group (Group D) and PPI-praziquantel combination group (Group E), of 10 mice in each group. Each mouse in groups B, C, D and E was infected with (30 ± 2) S. japonicum cercariae. Then, mice in groups D and E were given praziquantel by gavage at a dose of 500 mg/kg for successive two days on day 42 post-infection, while mice in groups C and E were given PPI by gavage at a dose of 400 mg/kg for successive 30 days on day 42 post-infection. Histopathological changes of hepatic tissues were observed using hematoxylin-eosin (HE) staining, and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN) were determined, while the activities of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), glutathione reductase (GSH-R) and glutathione (GSH) were detected in Mouse liver homogenates. The expression of transforming growth factor-beta (TGF-β) and alpha-smooth muscle actin (α-SMA) was quantified in hepatic tissues using immunohistochemistry, and the Nrf2 and Gsta4 gene expression was quantified using quantitative real-time PCR (qPCR) assay. Results Untreated mice presented typical pathological changes of schistosomal hepatic disorders, while PPI treatment effectively alleviated hepatic egg granulomas and collagen deposition. S. japonicum infection resulted in aggravation of hepatic lipid peroxidation, induction of oxidative stress, elevated serum MDA level and a reduction in the activity of GSH and antioxidant enzymes activities in mice. As compared to infected but untreated mice, PPI treatment suppressed hepatic lipid peroxidation, increased the GSH activity and restored the activity of antioxidant enzymes. In addition, PPI treatment inhibited the TGF-β signaling pathway and up-regulated the Nrf2 and Gsta4 gene expression. Conclusions PPI plays a critical role in the treatment of schistosomiasis-induced hepatic fibrosis. It may improve oxidative stress damages through up-regulating Nrf2 and Gsta4 gene expression, thereby suppressing the development of hepatic egg granulomas and hepatic fibrosis.
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Studies have shown that stromal interaction molecule 1(STIM1) is closely related to the development of tumors, and which is involved in the regulation of apoptosis, proliferation, migration and invasion in many human cancers. Blocking or knockdown of STIM1 can significantly inhibit the proliferation and migration of cancer cells. Elucidation of the regulatory mechanism of STIM1 in cancer cells will be helpful for the identification of new therapeutic targets. This paper reviews the mechanism of STIM1 molecule in different tumors and its clinical application.
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Objective To discuss the safety and efficacy of laparoscopic modified ventrifixation for women with uterine prolapse.Methods Thirty-seven women with Ⅱ-Ⅳ degree uterine prolapse were undergent laparoscopic modified ventrifixation in the the First Hospital of Qinhuangdao and Anzhen Hospital of Beijing from January 2008 to June 2012.The midpart of a noabsorble PROLENE soft mesh was sutured to the anterior cervical fascia.Two back ends of the mesh were passed through extraperitoneal channels through ligamentum latum uteri and sutured to the abdorminal wall to fix uterus.The effect and complications were observed.Results The objective and subjective success rate were 100％ and 91.9％ respcetively at 6 months after operation.All patients were followed up for 6 to 48 months.Prolapse recurrence rate was 10.8％ (4/37).Conclusion Laparoscopic modified ventrifixation is effective,safe and mini-invasive in the treatment of uterine prolapse.The surgery may be a satisfactory procedure for women with uterine prolapse hoping for uterine preservation.
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Forty batches of Lonicerae Japonica Fse i collected extensively and prepared as the test solution. Their chromatographic fingerprints and anti-influenza virus IC50 value (half maximal inhibitory concentration) were determined respectively. Then Unscrambler software was used, and spectrum-efficient correlation analysis was done for chromatographic fingerprints data and IC50 data by partial least squares regression method, to establish spectrum-efficient correlation model for anti-influenza virus of Lonicerae Japonicae Flos. Then the other 10 batches of Lonicerae Japonicae Flos were used to verify the model and explore the adaptability of this spectrum-efficient correlation model based on partial least squares regression method. The mathematical model obtained R2 of 0.969489 and RM-SEC of 0.070691 for calibration set; R2 of 0.959042 and RMSECV of 0.084005 for cross validation set. The verification experiment results showed that the relative error between the predicted values and measured values was within 10% in all 10 hatches, and within 5% in 80% of them. The results showed that the established spectrum-efficient correlation model could be used to evaluate the biological activity of anti-influenza virus of Lonicerae Japonicae Flos by determining its HPLC fingerprints.
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Antiviral Agents , Pharmacology , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacology , Flowers , Chemistry , Least-Squares Analysis , Lonicera , Chemistry , Molecular Structure , OrthomyxoviridaeABSTRACT
The theoretical basis of the alcohol precipitation process control was provided, the alcohol precipitation was optimized and the relationship equation was got. The monod glycoside, loganin and paeoniflorin were used as the evaluation indexes to determine the impact factors of alcohol precipitation techniques of Liuwei Dihuang decoction by the Plackett-Burman experimental design and the levels of non-significant factors were identified. Then, Box-Behnken response surface methodology was used to research and discuss the critical process parameters influence the effect of alcohol precipitation and draw interaction between key process parameters and the correlation equation with index components. Through the establishment and solving the quadratic regression model of composite score, the optimum preparation conditions of alcohol precipitation techniques of liuwei were as follows: stirring speed was 580 r x min(-1), standing time was 17 hours, alcohol concentration was 34%, the density of Liuwei Dihuang decoction was 1.13. The response surface methodology for optimized alcohol precipitation techniques of Liuwei Dihuang decoction is. reasonable and feasible.
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Chemical Precipitation , Chemistry, Pharmaceutical , Methods , Drugs, Chinese Herbal , Chemistry , Ethanol , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of different pH on rectum permeability of chlorogenic acid and geniposide.</p><p><b>METHOD</b>Four kinds of Reduning suppositories of different pH were separated and put into the rectum to study the suppositories in vitro and the content of chlorogenic acid and geniposide samples was determined by HPLC to calculate the permeation in 24 hours.</p><p><b>RESULT</b>With increase of pH within 2.5-7.4, the steady state flux of chlorogenic acid was increased, but the steady state flux of geniposidesamples was steady.</p><p><b>CONCLUSION</b>Adjusted the pH can increase the rectum permeability of active ingredients in Reduning auppositories.</p>
Subject(s)
Animals , Male , Rats , Chlorogenic Acid , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacokinetics , Hydrogen-Ion Concentration , Iridoids , Pharmacokinetics , Permeability , Rats, Sprague-Dawley , Rectum , Metabolism , Suppositories , PharmacokineticsABSTRACT
To determine the optimum process conditions for dry granulating technique of Qibai Pingfei granule, granule excipient type, rolling wheel speed and pressure and feeding speed were studied. Taking shaping rate at a time, moisture absorption and dissolubility as index, the type and amount of granule excipient were determined. In addition, taking shaping rate at a time as index, parameters of rolling wheel speed and pressure and feeding speed were researched through single factor test and response surface methodology. The optimum parameters were as follows: lactose as excipient, dry extract powder to excipient at 1:2, rolling wheel speed and pressure at 10.9 Hz and 6.4 MPa and feeding speed at 7.2 Hz. After validation of three batches pilot-scale production, the optimum processing parameters for dry granulating technique of Qibai Pingfei granule is reasonable and feasible, which can provide reliable basis for production.
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Drugs, Chinese Herbal , Powders , Technology, Pharmaceutical , MethodsABSTRACT
Objective To investigate the correlation between the deletion of exon 13 of hMSH2 mRNA in peripheral blood leu-kocyte and ISV12(-6) T>C polymorphism with sporadic colorectal cancer .Methods Total RNA and genomic DNA were extracted from peripheral blood of colorectal cancer patients and healthy controls .RT-PCR and PCR were used to amplified the mRNA and exon 13 of hMSH2 gene .The sequences of amplified hMSH2 cDNA ,ISV12(-6) T>C polymorphism and exon 13 sequence were confirmed by DNA sequencing .Results 23 of 23 (100% ) patients and 31 of 35 controls (88 .6% ,P>0 .05) were found to have an hMSH2 truncated transcript caused by a deletion of exon 13 .No deletions of exon 13 in hMSH2 gene were identified in genomic DNA .16 of 23 patients (69 .5% ) and 19 of 35 control (52 .3% ,P>0 .05) were found to have the T >C transition six bases up-stream of exon 13 of hMSH2 .Conclusion Deletion of hMSH2 mRNA exon 13 in peripheral blood leukocyte and the ISV12(-6) T>C polymorphism are common variants in population and have no correlation with sporadic colorectal cancer .The variant of splice site ISV12(-6)T>C is not a reason causing the deletion of hMSH2 mRNA exon 13 .
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This study was aimed to acquire the dynamic changes of chlorogenic acid and luteoloside of honeysuckle at different collecting periods in order to decide the best harvesting time of honeysuckle in Donghai County, Jiangsu Province. The content determination method used in the detection of chlorogenic acid and luteoloside of honeysuckle was from the 2010 edition of the Chinese Pharmacopoeia. The skills of HPLC fingerprint characteristic features, and the yield of pressed flowers were combined in the evaluation of honeysuckle at the three-green period, two-white pe-riod, great-white period, silver-flower period and the golden-flower period. The results showed that the content of honeysuckle at different blossoming stages had obvious changes in content of chlorogenic acid and luteoloside, as well as the pressed flower quality and yield of the flower. It was concluded that the optimal harvest time of honey-suckle was for the two-white period and the great-white period, which was consistent with the real origin.
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Objective To investigate the familial incidence of multiple primary carcinoma in Chinese hereditary nonpolyposis colorectal cancer patients (HNPCC) in Northeast China.Methods By family line investigation,multiple primary carcinoma (MPC) spectrum' s characteristics of 509 patients in 85 families registered in strict conformity with the HNPCC Amsterdam criteria Ⅱ were analyzed retrospectively.Results Of the 85 HNPCC families,multiple primary carcinoma developed in 55 patients in 25 families,among them 45 patients had metachronous carcinoma in 17 families,16 patients had synchronous carcinoma occurred in 12 families,6 patients with both synchronous carcinoma and metachronous carcinoma in 4 families.Conclusions Multiple primary carcinoma developed in significantly high incidence in Northeast China in Chinese hereditary nonpolyposis colorectal cancer patients,the most common MPC are colorectal cancer and endometrial cancer.
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Objective To investigate some improvements in the surgical techniques of adult-to-adult living donor liver transplantation( A-A LDLT) without the middle hepatic vein(MHV) for hepat-ic vein reconstruction. Methods The retrospective analysis was made on the clinical data of 11 recipi-ents who underwent the operation in A-A LDLT including the hepatic vein reconstructed in right liver lobe without MHV from June 2007 to January 2008. The key techniques included reconstructing out-flow of graft on shaping the tips of vena cava and right hepatic veins, cadaveric vein allografts stored in 4℃ UW solution within 7d being used for significant-sized hepatic vein reconstruction such as tributa-ries of the middle hepatic vein from V5, V8 and right inferior hepatic vein. Results 10 cases success-fully underwent reconstruction of outflow of graft on shaping the tips of vena cava and right hepatic veins and the outflow reconstruction ratio of V5, V8 and right inferior hepatic vein was 81. 8% (9/11), 7 one-vein reconstruction, 1 two-vein reconstruction and 1 three-vein reconstruction. 1 recipient died of renal failure and pulmonary infection 14 days after operation without venous outflow obstruc-tion. Doppler ultrasonography showed no thrombosis and the blood flowed smoothly in the right he-patic vein of other 8 recipients during the 9th to 15th mouth of follow-up. The cumulative patency rates of these 8 survivals for interposition vein grafts were 100% (11/11), 72. 7 %(8/11), 54. 5%(6/11) and 36. 5%(4/11) in 1, 3, 6 and 9 mouths, respectively. The regeneration of paramedian sectors was equivalent. Conclusion Shaping the tips of vena cava and right hepatic veins and using cadaveric vein allografts in adult-to-adult right lobe living donor liver transplantation for hepatic vein reconstruc-tion are both safe,simple and effective methods.This approach can be recommended.
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Objective To evaluate the changes of platelet-derived growth factor(PDGF) and vascular endo-thelial growth factor(VEGF) in infectious and autoimmunity diseaases. Methods The growth factors were measured by ELISA in 149 patients for seven times in different periods,and the levels and duration were compared. Results PDGF was (6.32±2.54) μg/ml and (2.57±1.65) μg/ml (P<0.05), and VEGF was (179.5±53.30) ng/ml and (221.4±70.04) ng/ml(P<0.05) in the two groups before treatment;There was statistical significance in dif-ferent time points between groups and within groups for PDGF(P<0.05) and significant difference for time chan-ging to the peak and changed duration between two groups for VEGF(P<0.05). For the two groups,the peak time of PDGF were (2.6±1.1) and (5.4±3.3) days;The peak time of VEGF were (2.4±0.7) and (7.2±3.3) days respectively(P<0.05 for each). However in the two groups, the change duration were (6.7±3.1) and (15.4±6.1) days for PDGF and (8.1±3.4) and (16.7±7.2) days for VEGF (P<0.05 for each). Conclu-sions There is significant difference for serum levels of PDGF and VEGF and the level change duration as well in the two groups, which maybe correlated with inflammatory nature and outcome.
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<p><b>OBJECTIVE</b>To explore the methods of the treatment and the principles of the prevention of bronchus-pleural fistula (BPF) after pneumonectomy.</p><p><b>METHODS</b>The clinical data of 15 cases of BPF after pneumonectomy in 815 lung cancer cases treated from July 1999 to June 2006 were analyzed retrospectively.</p><p><b>RESULTS</b>The occurrence rate of BPF after right pneumonectomy was 3.9% (12/310), higher than 0.6% (3/505) of left pneumonectomy (P < 0.01). The occurrence rate of BPF in cases with positive cancer residues in stump of bronchus was 22.7% (5/22), higher than 1.3% (10/793) of the cases with negative stump of bronchus (P < 0.01). The occurrence rate of BPF in the cases received preoperative radio- or chemotherapy was 5.0% (6/119), higher than 1.3% (9/696) of the cases received operation only (P < 0.05). There were no BPF occurred in the 76 cases whose bronchial stump were covered with autogenous tissues. All of the cases diagnosed as BPF were undertaken either closed or open chest drainage. Two cases were cured by thoracentesis aspiration and infusion antibiotics repeatedly. Two cases were cured by blocking the fistula with fibrin glue after sufficient anti-inflammatory treatment and hypertonic saline flushing. Six cases were discharged with a stable condition after closed drainage only. One case was discharged with open drainage for long time and 1 case was cured by hypertonic saline flushing after failure to cover the BPF using muscle flaps. Three cases died of multi-organs functional failure.</p><p><b>CONCLUSIONS</b>BPF are related to the bronchial stump management and positive or negative residue of tumor at the bronchial stump. Autogenous tissues covering of the bronchial stump is a effective method for decrease the rate of BPF and especially for those patients received preoperative radio- or chemotherapy and right pneumonectomy. It should be performed for early mild cases with repeated thoracentesis aspirations or blocking the fistula with fibrin glue together with antibiotics. Chest closed drainage immediately and flushing with hypertonic saline repeatedly are effective methods for BPF.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bronchial Fistula , Epidemiology , Therapeutics , Lung Neoplasms , General Surgery , Pleural Diseases , Epidemiology , Therapeutics , Pneumonectomy , Methods , Postoperative Complications , Therapeutics , Retrospective Studies , Treatment OutcomeABSTRACT
Objective To investigate the surgical treatment and perioperative care of the elderly patients with colorectal cracinoma.Methods The surgical treatment of 77 elder patients above 70 with colorectal carcinoma was analyzed.Results 54 patients were resected curatively,tumor resection rate 70%,12 patients were resected palliatively,9 cases in cecostomy,2 cases in by-pass operation.26 instances of postoperative complications happened.5 in death,with 6% mortality rate.Conclusion Allround preoperative preparation,strengthy perioperative care and correct operation procedure is a safety and effective way to raise tumor resection rate and reduce preexisting diseases for the elderly patients with colorectal carcinoma.