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ObjectiveTo explore the effect and possible mechanism of Shenqi Pills （肾气丸） on cognitive impairment and hippocampal glucose energy metabolism in type 2 diabetes mellitus （T2DM）. MethodsSixty C57BL/6 mice were randomly divided into control group， model group， rosiglitazone group and Shenqi Pills low-， medium- and high-dose groups， with 10 mice in each group. T2DM model was induced by a high-fat diet combined with intraperitoneal injection of streptozotocin in all the groups except for the control group. After successful modeling， the high-， medium-， and low-dose Shenqi Pills groups were given 2.08， 1.04， and 0.52 g/（kg·d） of Shenqi Pills granules by gavage respectively， while the rosiglitazone group was given 3 mg/（kg·d） of rosiglitazone tablets by gavage， and the control group and model group were gavaged with 10 ml/（kg·d） of distilled water， all for 8 consecutive weeks. The body weight and fasting blood glucose （FBG） level were recorded every two weeks. The Morris water maze test was performed on the 8th week of medication. After 8-week medication， oral glucose tolerance test （OGTT） and fasting insulin level were measured， hippocampal glucose energy metabolism-related products were quantitatively detected by liquid chromatography tandem mass spectrometry， and KEGG annotation analysis was performed. ResultsCompared to those measured at the same timepoints in the control group， the body mass on week 6 and 8， as well as the FBG level on week 2， 4， 6 and 8 in the model group increased； the blood glucose level at 0， 30， 60 and 120 minutes of the OGTT test increased， while fasting insulin level after 8-week medication decreased. The escape latency of the model group was significantly prolonged on the 3rd and 4th days， and the escape latency time increased， while the total swimming distance， platform quadrant residence time and the number of platform crossings decreased （P＜0.05 or P＜0.01）. Compared to those measured at the same timepoints in the model group， the body mass on week 6 in the low-dose Shenqi Pills group， on week 6 and 8 in the medium- and high-dose groups， and on week 8 in the rosiglitazone group were significantly reduced； the FBG levels in all the Shenqi Pills groups and rosiglitazone group on week 6 and 8 decreased， while fasting insulin levels increased. In the OGTT test， blood glucose in the medium-dose group of Shenqi Pills at all timepoints decreased； in the Morris water maze test， the escape latency period of the medium- and high-dose Shenqi Pills groups was shortened on the 3rd and 4th days， while the escape latency time was reduced， and the total swimming distance， platform quadrant residence time， and number of platform crossings increased in the medium-dose Shenqi Pills group （P＜0.05 or P＜0.01）.The medium-dose Shenqi Pills showed best effect， therefore it was selected for the targeted quantitative detection of metabolites. The medium-dose Shenqi Pills group could regulate the disorder of glucose metabolism in the hippocampus of T2DM mice， and 13 differential metabolites were found，up-regulating α-ketoglutarate and 3-phosphoglyceric acid， and down-regulating fumaric acid， glutamatic acid， lactatic acid， inosine， malic acid， adenine， fructose 1,6-diphosphate and others. KEGG annotation of differential metabolites suggested that Shenqi Pills was closely related to the regulation of glucose metabolism disorder and insulin resistance in the hippocampus region of T2DM model mice， as well as neurodegenerative diseases and ABC transport， hypoxia-inducible factor 1 （HIF）， forkhead transcription factor （FoXO） and cyclic adenosine monophosphate （cAMP） signaling pathways. ConclusionShenqi Pills can improve learning and memory abilities and cognitive impairment in T2DM mice， and may act its role by regulating glucose energy metabolism in the hippocampus of T2DM.
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Objective To investigate the changes in the expression of cold-inducible RNA-binding protein (CIRBP) in a radiation-induced lung injury model. Methods Thirty male C57BL/6 mice were randomly divided by body weight into control group (no intervention) and model group (single chest X-ray irradiation with a dose of 20 Gy to build a radiation-induced lung injury model). The mice were dissected five weeks after irradiation. Hematoxylin-eosin staining and Masson staining were used to observe the pathological changes of the lung tissue and the deposition of collagen fibers. Immunohistochemistry was used to measure the expression of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the lung tissue. qRT-PCR was used to measure the expression of CIRBP mRNA in the lung tissue. The expression of CIRBP protein in the lung tissue was determined by the immunofluorescence assay and Western blot. Results Compared with the control group, the model group showed significant pulmonary vascular congestion, significant inflammatory cell infiltration, significant thickening of some alveolar septa, significantly increased IL-6 expression [(129.41 ± 5.58) vs (187.22 ± 34.77), t = 3.179, P < 0.05], significantly increased TNF-α expression [(137.52 ± 23.53) vs (187.02 ± 19.16), t = 5.069, P < 0.05], significantly increased CIRBP mRNA expression [(1 ± 0.08) vs (1.97 ± 0.39), t = 3.45, P < 0.05], and significantly increased CIRBP protein expression [(9.32 ± 1.26) vs (14.76 ± 1.61), t = 3.751, P < 0.05], by the immunofluorescence assay; [(1.13 ± 0.17) vs (1.49 ± 0.14), t = 2.819, P < 0.05], by Western blot). Conclusion The expression of CIRBP is significantly increased in the radiation-induced lung injury model, which may be an important pro-inflammatory factor in radiation-induced lung injury.
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Objective:To evaluate the effects of rocuronium on limb ischemia-reperfusion injury in the patients undergoing total knee arthroplasty.Methods:Ninety patients of both sexes, aged 50-80 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with body mass index<30 kg/cm 2, scheduled for elective unilateral knee arthroplasty under general anesthesia combined with femoral nerve block from January 2019 to October 2019, were divided into 3 groups ( n=30 each) using a random number table method: normal saline group (group S), rocuronium 0.6 mg/kg group (group RL) and rocuronium 1.2 mg/kg group (group RH). Anesthesia was induced by intravenous injection of midazolam, etomidate, sufentanil and rocuronium 0.6 mg/kg (group RL) or 1.2 mg/kg (group RH) or equal volume of normal saline (group S). Mechanical ventilation was performed after placement of laryngeal mask to maintain P ETCO 2 35-45 mmHg.Femoral nerve block was performed under ultrasound guidance.Anesthesia was maintained by inhaling 1% sevoflurane and intravenous infusion of propofol and remifentanil to maintain entropy index at 40-60 during operation.Patient-controlled intravenous analgesia was performed with sufentanil after surgery to maintain visual analogue scale score ≤ 4 points.When visual analogue scale score was > 4, flurbiprofen axetil 100 mg was intravenously injected.The vastus medialis muscle at the edge of the incision was obtained at 60 min after inflation to determine the expression of dystrophin by immunohistochemistry.Arterial blood samples were collected immediately after inflation of the tourniquet, at 60 min after inflation, and at 5 and 30 min after deflation for determination of the serum malondialdehyde (MDA) concentrations by the thiobarbituric acid method.The effective pressing times of the analgesic pump and consumption of sufentanil and flurbiprofen axetil were recorded within 48 h after operation.The occurrence of responses to tourniquet and residual muscle relaxation during recovery from anesthesia, the first postoperative off-bed time and postoperative length of hospital stay were recorded.The thigh girth was measured before operation and at 24 and 48 h after operation, and the difference after and before operation was calculated.The range of motion of knees of the operated limb and tourniquet-related complications in the early postoperative period (3 days) and in the long-term postoperative period (3 months) were recorded. Results:Compared with group S, the expression of dystrophin in skeletal muscle was significantly up-regulated, the concentration of serum MDA was decreased at 30 min after deflation, and the difference in thigh girth at 24 and 48 h after operation was decreased in group RH, and the range of motion of knees was significantly increased at 3 days and 3 months after operation, and the first postoperative off-bed time was shortened in group RH and group RL ( P<0.05). Compared with group RL, the range of motion of knees was significantly increased at 3 days and 3 months after operation, and the first postoperative off-bed time was shortened in group RH ( P<0.05). There was no significant difference in the incidence of responses to tourniquet, postoperative length of hospital stay, effective pressing times of the analgesic pump, postoperative consumption of sufentanil and flurbiprofen axetil, and the incidence of tourniquet-related complications in the early and long-term postoperative periods among the three groups ( P>0.05). No residual muscle relaxation was found during recovery from anesthesia in three groups. Conclusion:Rocuronium 1.2 mg/kg can reduce limb ischemia-reperfusion injury in the patients undergoing total knee arthroplasty.
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Objective To study the clinical application effect of risk preventive nursing on children with pneumonia and their rehabilitation. Methods One hundred children with pneumonia admitted to Xiaoyi People's Hospital from March 2016 to March 2017 for treatment were enrolled. Among them, 45 cases from March the 1st to August the 1st in 2016 were treated with conventional nursing (conventional nursing group), while 55 cases from August the 2nd, 2016 to March the 1st, 2017 were taken cared by the risk prevention nursing (risk prevention group). The levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-8) were examined on admission and discharge in the two groups by enzyme-linked immunosorbent assay (ELISA); the time length for clinical symptoms to disappear, rehabilitation effect, hospitalization time, family members' satisfaction and incidence of complications were observed in the two groups. Results No statistically significant differences were detected in serum TNF-α, IL-6 and IL-8 levels between the two groups before nursing (all P > 0.05).The levels of inflammatory factors at discharge in both groups were lower than those at admission, the levels of TNF-α, IL-6 and IL-8 in the children of risk prevention group were significantly lower than those in the conventional nursing group discharge [TNF-α (μg/L): 16.54±7.13 vs. 22.78±9.12, IL-6 (ng/L): 9.25±5.48 vs. 15.11±7.01, IL-8 (μg/L): 4.08±2.16 vs. 7.69±3.56, all P < 0.05]. The duration for clinical symptoms to disappear and duration of hospital stay in the risk prevention group were significantly shorter than those in the conventional nursing group [cough stopping time (days): 6.87±2.54 vs. 10.35±3.08, duration of rale disappearance (days): 7.01±2.13 vs. 10.87±3.25, fever recovery time (days): 6.25±2.64 vs. 8.76±3.58, duration of asthma relief (days): 7.59±3.17 vs. 10.26±3.26, duration of hospital stay (days): 8.16±1.86 vs. 13.25±3.64, all P < 0.05]. The total effective rate and family members' satisfaction in the risk prevention group were significantly higher than those in the conventional nursing group [total effective rate: 94.54% (52/55) vs. 77.78% (35/45), family members' satisfaction: 96.36% (53/55) vs. 84.44% (38/45), both P < 0.05]. The incidence of complications in the risk prevention group was obviously lower than that in the conventional nursing group [5.45% (3/55) vs. 46.67% (21/45), P < 0.05]. Conclusion Risk prevention nursing can reduce the incidence of inflammatory reaction, improve the family members' satisfaction with nursing, and promote the children recovery process.
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Objective To investigate the feasibility of using optimized protocol of iodine contrast agent with fixed injection time in triple-rule-out CT examination of acute chest pain patients. Methods A prospective study was conducted. The patients who underwent triple-rule-out CT examination of acute chest pain at the Second Hospital of Shanxi Medical University from September 2017 to June 2018 were enrolled. According to the patient's body mass index (BMI), they were divided into BMI ≤ 23 kg/m2 group and BMI > 23 kg/m2 group. The patients in each group were subdivided into two subgroups according to the random number table, and they were given two iodine contrast injection protocols with fixed injection time (14 s). Protocol 1 was performed with 55 mL of total iodinated contrast media: iodinated contrast media was first injected at 5.0 mL/s for 8 s, followed by the same contrast media injection at 2.5 mL/s for 6 s, finally followed by injection of 40 mL of saline at a rate of 2.5 mL/s. Protocol 2 with 60 mL of total iodinated contrast media: iodinated contrast media was first injected at 5.0 mL/s for 10 s, followed by the same contrast media injection at 2.5 mL/s for 4 s, finally followed by injection of 40 mL of saline at a rate of 2.5 mL/s. The primary and objective evaluation was conducted on the image quality of the patients' blood vessels in different segments. The primary score, CT value and contrast-to-noise ratio (CNR) of the pulmonary artery, coronary artery, aorta and total effective radiation dose for the examination were recorded. Results A total of 92 patients were enrolled in the analysis. There were 44 patients in BMI≤ 23 kg/m2 group, in which 22 patients received in protocol 1 and protocol 2, 48 patients in BMI > 23 kg/m2 group, in which 24 patients in protocol 1 and protocol 2, respectively. There was no significant difference in the effective radiation dose between the two subgroups receiving different injection protocols in different BMI groups (mSv: 6.7±1.1 vs. 6.5±0.8 between protocol 1 and protocol 2 in BMI ≤ 23 kg/m2 group; 7.8±1.0 vs. 8.0±1.1 between protocol 1 and protocol 2 in BMI > 23 kg/m2 group, both P > 0.05). In BMI ≤ 23 kg/m2 group, the CT value, CNR and primary scores of pulmonary artery images in patients receiving protocol 2 were significantly higher than those receiving protocol 1 [CT value (HU): 584±110 vs. 472±86 for main pulmonary artery, 561±93 vs. 467±78 for left pulmonary artery, 555±91 vs. 472±83 for right pulmonary artery; CNR: 24.2±7.5 vs. 18.7±4.6 for main pulmonary artery, 23.2±6.8 vs. 18.6±4.8 for left pulmonary artery, 22.9±6.7 vs. 18.8±4.7 for right pulmonary artery; primary score:4.0 (4.0, 4.0) vs. 3.5 (3.0, 4.0), all P < 0.05]; and there was no statistically significant difference in the primary or objective evaluation of coronary artery or aortic image quality between the two protocols. In BMI > 23 kg/m2 group, the CT value, CNR and primary scores of coronary artery and aortic images in patients receiving protocol 2 were significantly higher than those receiving protocol 1 [CT value (HU): 369±63 vs. 315±61 for proximal right coronary artery (RCA), 388±63 vs. 323±63 for proximal left coronary artery (LCA), 328±83 vs. 272±51 for ascending aorta, 348±82 vs. 272±49 for aortic arch; CNR: 15.0±4.6 vs. 12.3±4.7 for proximal RCA, 15.7±3.8 vs. 12.8±5.2 for proximal LCA, 13.2±5.3 vs. 10.4±4.1 for ascending aorta, 14.1±5.3 vs. 10.4±3.9 for aortic arch; primary score: 4.0 (3.0, 4.0) vs. 3.0 (3.0, 4.0) for coronary, 4.0 (3.0, 4.0) vs. 3.0 (2.0, 4.0) for aorta; all P < 0.05]; and there was no statistically significant difference in the primary or objective evaluation of pulmonary artery image quality between the two protocols. Conclusions The effective radiation dose of triple-rule-out CT examination of acute chest pain is relatively low. The low-dose iodine contrast agent application program with fixed injection time can meet the needs of clinical diagnosis of triple-rule-out CT examination of acute chest pain patients. For patients with BMI ≤ 23 kg/m2, both protocols 1 and 2 can obtain excellent image quality; in order to avoid the influence of superior vena cava artifacts, protocol 1 is recommended. For patients with BMI > 23 kg/m2, application protocol 2 can obtain stable, excellent image quality that is more suitable for clinical applications.
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Lung cancer stem cells play an important role in chemoresistance and invasion and metastasis of lung cancer. Increasing evidence sustains that microRNAs(miRNAs)play a major role in regulating tumor angiogenesis,drug resistance and metastasis. Cancer stem cells are considered to be one of the reasons leading to cancer recurrence,metasta-sis and resistance to chemotherapy,therefore better understanding how miRNAs regulate gene expression in lung cancer stem cells will help identify cancer biomarkers and new therapeutic targets,and will help to develop better treatment strate-gies for lung cancer.
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Objective To investigate the effect of targeted silencing Notch1 on proliferation and apoptosis of human non-small cell lung cancer stem cells.Methods Lung cancer A549 cells and SPC-A-1 cells were selected and divided into control group,Nc-shRNA group and Notch1-shRNA group.The Nc-shRNA group was a negative control RNAi lentivirus group,and the Notch1-shRNA group was a Notch1 inhibitory RNAi lentivirus group.The lentiviral-mediated shRNA interference technology was used to target the silencing of Notch1.The silencing effect of Notch1 gene was verified by quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting.Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) and sarcosphere formation assay.Apoptosis was detected by Annexin V/7-AAD double staining.Western blotting was used to detect the expression of proliferating cell nuclear antigen (PCNA),B-cell lymphoma-2 (Bcl-2) and Notch1 downstream gene Hes-1.Results The results of qRT-PCR showed that the relative expression levels of Notch1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1 cells were 1.000 ± 0.000,0.937 ± 0.025,0.490 ± 0.036 and 1.000 ± 0.000,1.077 ± 0.070,0.373± 0.038,with statistically significant differences (F =359.707,P ＜0.001;F =210.455,P ＜0.001),further paired comparison,the relative expression of Notch1 in Notch1-shRNA group was significantly lower than that in Nc-shRNA group (all P ＜ 0.05).Western blotting showed that the expressions of Notch1 protein in A549 cells and SPC-A-1 cells were consistent with the mRNA results.MTT assay showed that the 24 h A values of A549 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.209 ± 0.005,0.219 ± 0.009,0.159 ±0.006,48 h A values were 0.293 ± 0.004,0.302 ± 0.004,0.205 ± 0.005,72 h A values were 0.450 ± 0.003,0.430 ± 0.012,0.348 ± 0.017,with statistically significant differences (F =79.487,P＜0.001;F =508.664,P ＜0.001;F =57.156,P ＜0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 24,48,72 h (all P ＜ 0.05).The 48 h A values of SPC-A-1 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.438 ±0.022,0.412 ± 0.015,0.364 ± 0.010,72h A values were 0.540 ± 0.016,0.519 ± 0.009,0.438 ± 0.019,with statistically significant differences (F =15.667,P =0.004;F =37.299,P ＜ 0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 48 h and 72 h (all P ＜ 0.05).The sphere sizes of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were (149.667 ± 6.506) μm,(136.667 ± 7.095) μm,(86.676 ± 7.638) μm,with statistically significant difference (F =65.940,P ＜ 0.001).The sphere sizes of the three groups in SPC-A-1 cells were (118.667 ± 6.658) μm,(128.000 ± 7.000) μm,(60.675 ± 4.509) μm,with statistically significant difference (F =105.372,P ＜0.001).Further paired comparison,the sphere size of Notch1shRNA group was significandy smaller than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).The apoptosis rates of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1cells were (0.489 ± 0.014)％,(0.633 ± 0.021)％,(1.683 ± 0.221)％ and (1.323 ± 0.194)％,(1.690 ± 0.188) ％,(3.017 ± 0.356) ％,with statistically significant differences (F =77.660,P ＜ 0.001;F=32.200,P =0.001),further paired comparison,the apoptosis rate of Notch1-shRNA group was significantly higher than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).Western blotting showed that the expressions of PCNA,Bcl-2 and Hes-1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were statistically significant (F =155.343,P ＜ 0.001;F =22.576,P =0.002;F =70.108,P＜0.001),and the expressions of PCNA,Bcl-2 and Hes-1 in the three groups in SPC-A-1 cells were statistically significant (F =49.419,P ＜0.001;F =28.090,P =0.001;F =12.040,P =0.007).Further paired comparison,the expressions of PCNA,Bcl-2 and Hes-1 in Notch1-shRNA group were significantly lower than those in Nc-shRNA group in the two kinds of cells,and the differences were statistically significant (all P ＜0.05).Conclusion Targeted silencing of Notch1 can reduce the proliferation activity of lung cancer stem cells and promote apoptosis,which may be related to the down-regulation of its downstream gene Hes-1.
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Notch signaling pathway is involved in the abnormal differentiation and self-renewal of lung cancer stem cells.The further studies for the roles of Notch signaling pathway in the regulation of lung cancer stem cells are expected to find new targets in the diagnosis and treatment of lung cancer.Inhibitors of the Notch signaling pathway may be effective in the treatment of lung cancer.Lung cancer stem cells are thought to be a major cause of recurrence of lung cancer,therefore,targeted therapy for lung cancer stem cells may be more effective than treatment for the entire tumor.
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Objective:To investigate the regulatory mechanism of PESV on tumor-infiltrating natural killer ( NK) cells in a mice model with H22 orthotopic transplantation tumor .Methods:Suspensions of H22 cells were injected into the lobe of liver on C 57BL/6 mice for establishing liver orthotopic transplantation tumor model ,then the mice were randomly divided into four groups:normal group , control group ,PESV low dose group ( PESV-L ) and PESV high dose group ( PESV-H ) .Mice were either sacrificed for mechanistic studies or survival followed 14 days of therapy.The volume and weight of the tumor were measured .The proportion of infiltrating NK cells was measured by flow cytometry and the expression of NK 1.1(NK) cells was investigated by immunohistochemistry method .The expression of perforin and granzyme B were further investigated by real-time PCR.Results: In contrast to control group , the tumor inhibition rate was 15.38%and 30.77% in PESV-L group and PESV-H group respectively.The survival showed that PESV-H could significantly prolong the survival time of mice ,and life extension rate was 34.06%,(P<0.05).Histological analysis revealed significant pleomorphism of the neoplastic cells and invasive extendion in control group ,while there were more necrosis and less degree of atypia in PESV-L and PESV-H.The level of tumor-infiltrating NK cell was significantly higher in PESV-H than in tumor-bearing control group [(5.91±0.49)%vs.(3.69±0.50)%,P<0.05],and NK cells were infiltrating in peritumoral lesions.The mRNA of perforin and granzyme B in PESV-H were respectively 3.62 and 5.82 times than that of control group ( P<0.05 ) .Conclusion: These findings suggest that the treatment of PESV might increase the infiltration of natural killer cells in the orthotopic transplantation tumor and contribute to NK cells migration to the tumor , which induct and maintain the activities of natural killer cells against tumor cells by expressing perforin and granzyme B in vivo .
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Objective To observe the clinical effect of preoperative intranasal dexmedetomidine on sedation and preventing postoperative agitation in children inguinal hernia operation .Methods Forty children patients ,aged 1-s5 years old ,ASA Ⅰ ,were selected and equally randomized into 2 groups ,receiving intranasally dexmedetomidine 1 μg/kg(group Dex) and the same volume of normal saline (group NS) at preoperative 30 min respectively .The sedation score ,emotion score when separating from their par‐ents ,mask inhalation induction resistance score were evaluated at 30 min after medication ,furthermore the agitation scores were ob‐served and recorded at postoperative 30 min ,1 ,2 h .Results The sedation score ,emotion score when separating from their parents and mask inhalation induction resistance score in the Dex group were superior to the NS group ,moreover the agitation score at each points were lower than those in the NS group(P<0 .05) .Conclusion Preoperative intranasal dexmedetomidine can be safely used in children inguinal hernia operation ,has better sedative effect ,meanw hile reduces the postoperative agitation occurrence .
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Objective To investigate the effects and the mechanism of sorafenib on hepatocellular car-cinoma growth and vasculogenic mimicry (VM)in mice.Methods A subcutaneous implantation mouse model of human hepatocellular carcinoma (HCC)HepG2 cells were established.Mice inoculated with HepG2 cells were randomly divided into the treatment group (sorafenib 30 mg·kg -1 ·d -1 )and the control group by using of paired comparison method.Growth of established tumor xenografts was monitored at least twice weekly by vernier caliper measurements.VMwas assessed by immunohistochemical assay and periodic acid schiff reaction (PAS)histochemical double-staining.The expressions of HIF-1 α,VEGFA,VEGFR-1 and MMP-2 in tumor tissues were also assessed by immunohistochemical assay,Western blotting and real-time quantitative PCR. Results The tumor volume in the sorafenib group was obviously decreased compared with the control group (809.69 mm3 ±208.71 mm3 vs 1 678.00 mm3 ±31 3.29 mm3 ),with a statistically significant difference (t =6.1 03,P =0.030).Haematoxylin and eosin (HE)staining showed that tumor tissues treated with sorafenib were characterized by obvious necrosis,but there were not the same cases in the control group.Sorafenib group significantly reduced the number of tumor functional vessel in HepG2 xenografts compared with the control group,as assessed by tumor vasculature uptake of DiOC7 (4.77 ±0.1 5 vs 8.44 ±0.68,t =9.1 92,P =0.01 3).The number of VMwas significantly decreased by sorafenib (1 .04 ±0.46 vs 2.66 ±0.42,t =4.51 0, P =0.041 ).Relative to controls,CD31 -positive vessels decreased after treatments (3.42 ±0.1 0 vs 1 .26 ± 0.1 4),with a statistically significant difference (t =21 .580,P =0.002).Compared with the control group, the protein levels of HIF-1 α(0.65 ±0.03 vs 1 .00 ±0.00),VEGFA (0.51 ±0.02 vs 1 .00 ±0.00), VEGFR-1 (0.45 ±0.04 vs 1 .00 ±0.00)and MMP-2 (0.69 ±0.02 vs 1 .00 ±0.00)were significantly decreased in the sorafenib group (t =1 9.650,P =0.003;t =40.493,P =0.000;t =23.429,P =0.002;t =26.071 ,P =0.002).Compared with the control group,the mRNA levels of HIF-1 α(0.78 ±0.05 vs 1 .00 ±0.00),VEGFA (0.52 ±0.05 vs 1 .00 ±0.00),VEGFR-1 (0.45 ±0.02 vs 1 .00 ±0.00)and MMP-2 (0.71 ±0.02 vs 1 .00 ±0.00)were also significantly decreased in sorafenib group (t =6.840,P =0.021 ;t =27.71 0,P =0.001 ;t =62.740,P =0.000;t =23.850,P =0.002).Conclusion Sorafenib can inhibit the tumor growth and VMchannels formation,which may be related with the HIF-1 αand VEGFA /VEGFR-1 signa-ling pathway.
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The molecular mechanism of autophagy is complicated and it always affects the occurrence and development of tumor in many different degrees,so it is the new hot spot in pathological study area.The angiogenesis includes physiological vascularization and pathological vascularization,and the pathological new vessels which provide nutrition for the continuous growth of tumors have a close relationship to the development and metastasis of tumor.The process of angiogenesis can be affected by regulating the autophagy of vascular endothelial cells,which provides a new method for tumor therapy.
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Cell autophagy is a lysosome dependent degradation pathway which is characterized by cytoplasm vacuoles.It is important to maintain cell internal environment.The abnormal autophagy is associated with oncogenesis closely.The tumor stem cells which rarely exist in tumor tissues have stem cell properties.They can induce tumor generation and also play an important role in the development,metastasis and recurrence of tumor.The regulation of autophagy in tumor stem cells is the research hotspot,which will provide a new theoretical foundation for the clinical treatment of tumors.
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<p><b>OBJECTIVE</b>To investigate the mechanisms for inhibition effects of PESV on proliferation of non-small cell lung cancer cell line A549.</p><p><b>METHOD</b>MTT was used to observe cell growth and proliferation of A549 at different concentrations of PESV. Flow cytometry (FCM) was applied to analyze cell cycle distribution. Immunocytochemistry and western blot assay was recruited to detect the expression of VEGF, HIF-1alpha, PTEN after the intervention of PESV.</p><p><b>RESULT</b>A549 cells may be arrested mainly in G0/G1 phase and cell proliferation was significantly inhibited (P < 0.01) after PESV intervention in a certain range of concentration. PESV can significantly reduce the expression of HIF-1alpha,VEGF and increase the expression of PTEN.</p><p><b>CONCLUSION</b>PESV can block cell cycle and inhibit angiogenesis directly to inhibit cell proliferation of non-small cell lung cancer cell line A549 mainly through reducing the expression of HIF-1alpha, VEGF and increasing the expression of PTEN.</p>
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Gene Expression Regulation, Neoplastic , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , PTEN Phosphohydrolase , Metabolism , Peptides , Pharmacology , Scorpion Venoms , Chemistry , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
ObjectiveTo explore the effects and underlying mechanisms of acid sensing ion channels (ASICs) on pain behavior in a rat model of post-incision pain.MethodsFifty-eight adult male Sprague Dawley rats were used in this study,four rats were used for immunofluorescence test,thirty rats were employed for pain behavior test,and twenty-four rats were used for Western blot.Rats used for pain behavior test and Western blot were randomly divided into 3 groups:control group ( C group),incision pain model group ( I group) and amiloride group (A group).Plantar skin of rats in A group were infiltrated with 20 μl(200 μg)amiloride solution.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency(PWTL) of all rats in pain behavior test was tested at 24 h preoperative,2 h,4 h,8 h,12 h,24 h postoperative.Western blot was tested at 4 h postoperative.ResultsImmunofluorescence test displayed ASIC3 was expressed in plantar skin of all rats.The basal level of PWMT and PWTL of all rats in three groups was C group( (23.15 ± 5.10) g,( 11.32 ± 1.21 ) s),I group ( (23.26 ± 5.69) g,( 11.75 ± 2.01 ) s),A group ( (23.63 ± 4.96 ) g,( 11.47 ± 1.96) s) respectively,which was no significantly difference (P ＞ 0.05 ).PWMT and PWTL of I group and A group was significantly lower than that of C group at all time points postoperative (P ＜ 0.05) ; PWMT and PWTL of A group was at 2 h( ( 13.75 ±3.25)g,(9.96±1.32)s),4h((14.05±3.75)g,(9.17±2.11)s),8 h((9.75 ±2.74)g,(8.11 ±1.22)s)postoperative,which was significantly higher than that of I group (P ＜ 0.05 ).Compared with that of C group,the level of pERK1/2 expression was significantly increased in I group at 4 h postoperative (P ＜ 0.05 ),which could be inhibited by amiloride local infiltration (P ＜ 0.05 ).ConclusionASIC3 can mediate incision pain in a rat model of post-incision pain,through pERK1/2 signaling pathway,which can be inhibited by amiloride.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effects of polypeptide extract from scorpion venom (PESV) alliance with chemotherapy on angiogenesis of Lewis lung carcinomas (LLC) and its mechanism.</p><p><b>METHOD</b>LLC cells suspension (4 x 10(6) cells/mL) were subcutaneously injected into 54 C57BL/6J mice in right armpits. Then the tumor-bearing mice were randomly divided into three groups: the control group, the chemotherapy group and the PESV group. Cyclophosphamide was used to establish the model of cancer. Chemotherapy and PESV were added to the PESV group. Every 7 days, 6 mice of each group were executed, and the experiments were carried out for 28 days. The tumor volume and inhibitory rate were determined. Immunohistochemistry and RT-PCR were used to determine the expression of factor VIII, alpha-SMA, Dll4 and Notch1 in tumor tissue. Correlation analysis was used to identify the relationship of factor VIII and calculate microvessel density (MVD), alpha-SMA and vascular maturity.</p><p><b>RESULT</b>The inhibitory rate of PESV was 42.21%. Comparing with the chemotherapy group, the expression of tumor factor Dll4 and Notch1 in the PESV group were decreased significantly (P < 0.05). The expression of factor VIII and alpha-SMA in the chemotherapy group is lower than the control group (P < 0.05), while it's higher when compared with the PESV group (P < 0.01). Expression of Dll4 and Notch1 in the chemotherapy group at the 28th day were higher than the control group (P < 0.05), and the expression in the PESV group at the 21st day were significantly lower than the chemotherapy group (P < 0.05).</p><p><b>CONCLUSION</b>PESV could inhibit the angiogenesis of LLC. It might be attained by decreasing the level of angiogenic factors, that are factor VIII, alpha-SMA, Dll4 and Notch1 in tumor microenvironment.</p>
Subject(s)
Animals , Male , Mice , Antineoplastic Agents , Blood , Chemistry , Therapeutic Uses , Carcinoma, Lewis Lung , Drug Therapy , Immunohistochemistry , Mice, Inbred C57BL , Neovascularization, Pathologic , Drug Therapy , Peptides , Chemistry , Therapeutic Uses , Reverse Transcriptase Polymerase Chain Reaction , Scorpion Venoms , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To study the expression of HIF-1alpha and SDF-1/CXCR4 in repopulating H22 tumor tissue and the mechanism of angiogenesis of polypeptide extract from scorpion venom (PESV) during chemotherapy treatment.</p><p><b>METHOD</b>The expression of HIF-1alpha and SDF-1/CXCR4 in H22 tumor tissue was monitored by immunohistochemistry, and the expression level was determined by Qwin V3 image analyzing software. The correlation between HIF-1alpha and SDF-1 was analyzed. SDF-1 content was detected by ELISA.</p><p><b>RESULT</b>HIF-1alpha expression was found no difference in model group between 14 d and 21 d, and up-regulated in 28 d. There was no change of HIF-1alpha expression was observed in low-dose PESV group. In high-dose PESV group, the level of HIF-1alpha expression was high in 14 d and low in 21 d. ELISA detecting showed SDF-1 content increased slowly from 14 d to 21 d, highly from 21 d to 28 d. But in high-dose PESV groups, the content increased slowly all the time. The immunohitochemistry method got the same result with ELISA. Correlation analysis showed r = 0.805. CXCR4 expression down-regulated in two PESV treated groups, and no difference was found between these two groups.</p><p><b>CONCLUSION</b>HIF-1alpha and SDF-1 participated in VEGF expression and angiogenesis in tumor tissue during chemotherapy, while PESV could inhibit the expression of HIF-1alpha and SDF-I.</p>
Subject(s)
Animals , Mice , Cell Line, Tumor , Chemokine CXCL12 , Metabolism , Down-Regulation , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Peptides , Pharmacology , Receptors, CXCR4 , Metabolism , Scorpion Venoms , Chemistry , Pharmacology , Scorpions , Chemistry , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To observe the inhabitive effect and mechanism of polypeptide extract from scorpion venom (PESV) on repopulation in H22 tumor cell during chemotherapy.</p><p><b>METHOD</b>H22 tumor cells were injected into 96 mice subcutaneously, then mice were divided into 4 groups radomly: Model, low-dose-PESV, high-dose-PESV, and control. Reppulation model was established by 5-Fu treating mice with H22. Four groups was treated differently, 6 mice of each group was sacrificed every 7 days, measured tumor volume twice one week. The expression of vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA), CD105 microvessel density (CD105-MVD) and platelet derived growth factor (PDGF) in H22 tumor issue was observed by using immunohistochemistry and grey analysis, the relation of VEGF and MVD was affirmed by correlation analysis.</p><p><b>RESULT</b>In control group tumor volume of H22 increased quickly in 13-24 day, and all mice died before 27 day. In model tumor volume increased quickly before 17, in 17-22 day slowly, after 22 day quickly again, and all the mice died before 31 day. In low and high dose PESV, tumor volume added slowly, and only in 17 day there was significant difference between these two groups. Immunohistochemistry showed, PCNA expression of model group in 31 day was higher than in 21, 28 day, the expression level of high and low PESV group was lower than model group all the time, only in 17 day there was significant difference between high and low PESV group. Immunohistochemitry showed, compared with high and low dose PESV group, CD105-MVD of model group was higher in 21, 28 day (P < 0.05) and in 35 day (P < 0.01), and no difference was found between high and low dose PESV. VEGF expression of model group in 35 day was higher than in 21, 28 day (P < 0.01), and model group higher than high and low dose PESV in 21, 28, 35 day. The expression of PDGF in model decreased gradually, in high and low dose PESV, the expression was lowest in 21 day. In day 35 high dose PESV higher than low dose PESV. There was positive correlation (r = 0.669) between VEGF expression and CD105-MVD.</p><p><b>CONCLUSION</b>PESV can inhabit repopulation of H22 tumor cell during chemotherapy, and the mechanism maybe is through anti-angiogenesis and nomalizing tumor vessels.</p>
Subject(s)
Animals , Mice , Angiogenesis Inhibitors , Therapeutic Uses , Antigens, CD , Metabolism , Carcinoma, Hepatocellular , Drug Therapy , Metabolism , Cell Line, Tumor , Immunohistochemistry , Liver Neoplasms , Drug Therapy , Metabolism , Peptides , Therapeutic Uses , Platelet-Derived Growth Factor , Metabolism , Proliferating Cell Nuclear Antigen , Metabolism , Scorpion Venoms , Chemistry , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effects of polypeptide extract from scorpion venom (PESV) on immune escape of Lewis lung carcinomas (LLC) and its mechanism.</p><p><b>METHOD</b>Forty C57BL/6J mice were inoculated with LLC cells suspension (1 x 10(7) cells/ mL) in right armpit subcutaneously. The tumor-bearing mice were randomly divided into two groups: the control group and the PESV group. PESV was intragastrically subjected to the mice of the experimental group for 18 days. The tumor volume and tumor inhibitory rate were determined. The expression levels of VEGF,TGF-beta1 and IL-10 in tumor microenvironment were determined by immunohisto-chemistry-staining and ELISA. Surface co-stimulatory molecules CD80 and CD86 of tumor infiltrating dendritic cells (DC) were determined by immunohistochemistry-staining and flow cytometry.</p><p><b>RESULT</b>The growth inhibitory rate of PESV was 56. 60%. The expression levels of VEGF,TGF-beta1 and IL-10 were decreased in tumor and serum, while the expression of co-stimulatory molecules CD80 and CD86 on DC were increased in tumor. Compared with the control group, the differences were all significant (P < 6.05).</p><p><b>CONCLUSION</b>PESV was effective in recovering immuno-surveillance and intervening immune escape of lung cancer through multi-pathway. And its effects might be attained by decreasing the level of VEGF, TGF-beta1 and IL-10 in tumor microenvironment and increasing the expression of co-stimulatory molecules CD80 and CD86 on DC.</p>