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Objective To establish the spatial course of distal tubule and afferent arterioles after macula densa, and to locate and detect the proteins in the adjacent parts by using three-dimensional visualization technology of microstructure. Methods C57 BL/6J mice were fixed by perfusion and embedded in epon 812. Tissue blocks were cut perpendicular to the longitudinal axis of the kidney. And a total of 720, 2. 5 μm-thick consecutive sections were obtained from the renal capsule to the outer stripe of the renal outer medulla. After aligning the digital microscopic images through computer registration procedures, the tubules and vessels were traced by 3D reconstruction program edited by C Language. Selecting the tissue sections of the contact site and applying the improved immunoperoxidase staining method to detect H
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Aim To discuss the mechanism of Lurong Dabu Decoction on cough variant asthma. Methods Guinea pigs were divided into normal group(CON), model group(OVA), Lurong Dabu Decoction high-dose group(HIGH),low-dose group(LOW), and dexamethasone group(DEX)at random. The CVA model was established by smoking plus injection of OVA, aluminum hydroxide solution and nebulized inhalation to stimulate cough. Gguinea pigs were dissected 24 hours after the last challenge to obtain alveolar lavage fluid(BALF)and lung tissues. Immunoadsorption(ELISA)method was applied to detect the types of inflammatory cells and the content of inflammatory cytokines in BALF; HE and Masson staining of the middle lobe of the left lung were used to observe the pathological changes in lung tissues; immunohistochemical staining was used to observe TLR4 and WNT-5A protein expression and distribution of lung tissues; the protein extracted from the upper lobe of the left lung was used to measure the level of TLR4 and WNT-5A protein in lung tissues by Western blot; immunofluorescence was employed to measure the fluorescence intensity of TLR4 and WNT-5A in lung tissues; flow cytometry was used to detect IL-4 and IFN-γ in guinea pig lung tissues. Results Lurong Dabu Decoction could improve guinea pig airway inflammation, inhibit collagen fiber deposition, reduce the content of IL-4, IL-5, and IL-13 in BALF, and inhibit the protein expression of TLR4 and WNT-5A in lung tissues and increase IFN-γ levels in lung tissues while decreasing IL-4 levels. Conclusion Lurong Dabu Decoction may inhibit the occurrence of CVA through TLR4/WNT-5A signaling pathway.
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Methods The model of heart failure after myocardial infarction was established by left coronary artery liga-tion in rats. Two weeks after modeling, all rats were randomly divided into model group, LGZGD group, and captopril group. Meanwhile sham operation group was set up. The rats were given continuous intragastric administration with drug or distilled water for 28 days, once a day. The behavioral signs of rats in each group were observed. The cardiac function of rats in each group was examined by echocardiography. Serum BNP and NT-ProBNP content were detected by enzyme-linked immunoassay; The changes of myocardial his-topathological and collagen fibers in rats were detected using sirius staining. The contents of oxidative stress index including ROS, SOD in myocardial tissue of rats in each group were observed by DCFH-DA fluorescent probe and Enzyme-linked immunoassay. The ultra-structure of mitochondria was observed by transmission electron microscopy. Expressions of apoptotic proteins ( mitochondrial CytC, cytoplasmic CytC) were detec- ted by Western blot. Expression of proteins related to the Nrf2/BNIP3 pathway were examined by immunoflu-orescence and Western blot. Results LGZGD could significantly improve the cardiac function of rats, reduce the contents of BNP and NT-ProBNP, inhibit the excessive deposition of collagen in myocardial interstiti-um, reduce ROS, increase the content of SOD, improve mitochondrial structure damage, up-regulate the expression of Nrf2 and nuclear translocation, and reduce the expression of BNIP3. Conclusions LGZGD can inhibit the ventricular remodeling and prevent the occurrence of heart failure after myocardial infarction. Its pharmacological effects are mainly related to regulating the Nrf2/BNIP3 pathway, activating Nrf2, promoting its nuclear transfer, and further down-regulating BNIP3 , protecting mitochondrial function, and reducing cardiomyocyte apoptosis.
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Objective: To compare the effects of artesunate (Art) and fuzheng huayu decoction on mitochondrial autophagy in the treatment of schistosomiasis liver fibrosis. Methods: Eighty C57BL/6 female mice were randomly divided into healthy control group, infection group, Art treatment group and Fuzheng Huayu Decoction treatment group, with 20 mice in each group. Mice in the infection group and treatment group were infected with 16 Schistosoma japonicum cercariae. After 6 weeks, praziquantel (300 mg/kg) was used for 2 days to kill the worms. The Art treatment group was treated with intraperitoneal injection of 100 mg/kg/day, while the Fuzheng Huayu Decoction treatment group was fed 16g of fuzheng huayu decoction per 1kg per day. After 6 weeks, fresh liver tissues of the four groups were collected. Masson staining and Western blot were used to observe the succinate dehydrogenase subunit A (SDHA) and malate dehydrogenase (MDH2), citrate synthase (CS), ketoglutarate dehydrogenase (OGDH), and target of rapamycin 1 (mTORC1) pathway involved in mitochondrial tricarboxylic acid cycle in liver tissues. The relative expression levels of adenylate activated protein kinase (AMPK) and mitochondrial autophagy pathway kinase (PINK1) were detected. Liver tissue samples were extracted from each group to detect the mitochondrial oxygen consumption rate. Two-way ANOVA was used to compare the significance and difference between two sets of samples. Results: Masson staining showed that the infection group mice had significantly higher liver fibrosis area than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group mice had lower liver fibrosis area than the infection group. Western blot analysis showed that the infection group (0.82 ± 0.05) had significantly lower relative expression of SDHA protein than the healthy control group (1.00 ± 0.05) (t = 11.23, P = 0.0035), while the Art treatment group (0.73 ± 0.05) had significantly higher relative expression of SDHA protein than the infection group (t = 10.79, P = 0.0073). However, there was no significant change in Fuzheng Huayu Decoction treatment group (0.98±0.05) (t = 1.925, P = 0.1266). The relative expression of p-AMPK protein was significantly higher in the infection group (1.15 ±0.05) than in the healthy control group (0.98 ± 0.07, t = 12.18, P = 0.0029), and the expression of p-AMPK in the Art treatment group (0.50 ± 0.05) was significantly lower than the infection group (t = 11.78, P = 0.0032). The relative protein expression of AMPK was significantly lower in the infection group (0.80 ± 0.05) than in the healthy control group (1.00 ± 0.05, t = 10.53, P = 0.0046). The expression of AMPK was significantly lower in the Art treatment group (0.54 ± 0.05) than in the infection group (T = 13.98, P = 0.0036). The relative expression of p-mTORC1 protein (0.93 ± 0.08) was not significantly different in the infection group than in the healthy control group (t = 2.28, P = 0.065), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1 protein than the infection group (t = 10.58, P = 0.029). The expression of p-mTORC1/ m-TORC1 was not significantly different in the infection group (0.98 ± 0.03) than in the healthy control group (0.97 ± 0.03, t = 0.98, P = 0.085), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1/ m-TORC1 than the infection group (t = 14.58, P = 0. 009). The relative protein expression of PINK1 was significantly lower in the infection group (0.55 ± 0.05) than in the healthy control group (1.00 ± 0.03, t = 13.49, P = 0.0011), while the Art treatment group (1.21 ± 0.05, t = 9.98, P = 0.0046) and Fuzheng Huayu Decoction treatment group (1.31 ±0.35, t = 6.98, P = 0.027) had significantly higher relative protein expression of PINK1 than the infection group. Mitochondrial function tests showed that after adding substrate complex II, the oxygen consumption of the infection group was lower than the healthy control group, while the Art treatment group and the Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. The oxygen consumption was significantly lower after adding the substrate complex III in the infection group than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. Conclusion: Art can alleviate schistosomiasis liver fibrosis by inhibiting AMPK/mTORC1 signaling pathway activity and enhancing mitochondrial oxygen consumption, autophagy and SDHA expression.
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Animals , Female , Mice , Artesunate , Drugs, Chinese Herbal/therapeutic use , Liver Cirrhosis/drug therapy , Mice, Inbred C57BL , Mitochondria , SchistosomiasisABSTRACT
KRAS is one of the most frequently mutated human oncogenes. In spite of mounting efforts on the development of direct or indirect inhibition targeting KRAS, little has been achieved because of insurmountable difficulties, titling KRAS "undruggable". Recently, subtype-specific inhibitors have shown great hope. Some KRASG12C inhibitors have entered clinical trials, including adagrasib and sotorasib, and have shown preliminary clinical effectiveness. Experiences from the inhibitors targeting the downstream factors of RAS pathways show that the anticancer activity of these drugs will be limited due to the development of drug resistance. Preclinical studies of KRASG12C inhibitors have revealed that the application of these agents might be hampered by the drug resistance issue. The current review aims to describe the current status of KRASG12C inhibitors, and discuss the mechanisms underlying KRASG12C inhibitor resistance, so as to provide the clues for the combat of drug resistance.
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Objective To analyze the influence of plaque classification and bifurcation angle on hemodynamics in coronary artery, so as to further discuss the influence on vulnerable atherosclerotic plaques. Methods Based on average geometric parameters of human coronary bifurcation vessels, the model of fluid-solid interaction for coronary bifurcation vessels with different plaque classifications and vessel bifurcation angles was constructed, and the distributions of blood flow velocity, pressure and shear stress at critical positions were investigated. Results The upstream shoulder of the plaque was the site with the highest shear stress on plaque surface, which was prone to ulceration or rupture with further growth. When there were plaques on one side of the bifurcation vessels, the shear stress at the carina of bifurcations was greater than that at the bilateral plaques. The pressure and shear stress at the carina of bifurcations gradually increased as the bifurcation angle decreased. Conclusions When there are plaques on one side of the bifurcation vessels, the probability of ulceration or rupture is greater. The presence of plaque in main vessels can promote formation and growth of the plaque at bifurcations. The inner wall of blood vessels at the carina of bifurcations is more easily damaged in the case of small angle vessels. The results can provide theoretical references for the design and optimization of vulnerable atherosclerotic plaque treatment.
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Objective To describe quantitatively the development of the capillary loop stage glomerulus (capG) with respect to the volume density of capillaries in the glomerulus based on the morphogenesis of the kidney. Methods The kidneys were obtained from mice at various developing time points and prepared for paraffin sections. The volume density of CD34 positive endothelial cells and surrounded capillary lumen in glomeruli was measured using a combination of immunohistochemical staining and the stereological grid system. Results The capG was divided into early, middle, and late phases, and middle phase capG was subdivided into early-middle and late-middle phases, according to the morphology of developing glomeruli and the arrangement of podocytes. As result, the volume density of capillary loops in early phase capG could not be measured due to the complex "glomerular" shape. The volume density of capillary loops increased from (35.95±6.45)% in the early-middle phase capG, to (58.36±6. 30) % in the late-middle phase capG, and to (79.89± 5.21) % in the late phase capG, compared to (93.61 ±1.96) % in the mature glomerulus. Furthermore, the volume density of capillary loops remained constant at same stage even though at different developmental time points. Conclusion This study demonstrated a significantly increased volume density of capillary loops with the kidney development. In addition, the results provide a descriptive and reliable parameter for the evaluation of glomerular development.
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This study was designed to explore the protective effect and underlying mechanism of catalpol on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD). High fat diet (HFD) was used to establish NAFLD model in the in vivo experiment, and the procedures of the experiments and animal care protocol were approved by the Animal Care and Use Committee of Jianghan University. Human liver cancer cell line HepG2 was treated with palmitate (PA) to establish a lipid toxicity model in the in vitro experiments. The results showed that catalpol significantly decreased the contents of serum total glyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), and aspartate transaminase (AST) in HFD-fed mice. Results of TUNEL staining and flow cytometry analyses revealed that catalpol significantly inhibited hepatocytes apoptosis in HFD-fed mice and PA-treated HepG2 cells. Moreover, catalpol treatment significantly reduced the endoplasmic reticulum stress-related protein expression levels of binding immunoglobulin protein (BiP), phosphorylated PKR-like endoplasmic reticulum kinase (p-PERK), inositol-requiring kinase 1α (IRE1α), and transcriptional factor activating transcription factor 6 (ATF6), and apoptosis-related protein expression levels of C/EBP homology protein (CHOP), phosphorylated c-Jun N-terminal kinase (p-JNK), and cleaved cysteinyl aspartate specific proteinases (caspases)-12, -9, and -3 in HFD-fed mice and PA-treated HepG2 cells. Furthermore, endoplasmic reticulum stress agonist tunicamycin (TM) significantly reversed the inhibitory effect of catalpol on protein expression levels of BiP, p-PERK, IRE1α, and ATF6, subsequently the inhibitory effect of catalpol on expression levels of CHOP, p-JNK, Bcl-2, Bax, and cleaved caspases (-12, -9, and -3) was also attenuated in PA-treated HepG2 cells. Taken together, these findings demonstrated that catalpol could inhibit hepatocytes apoptosis and had a significant protective effect on liver injury, and its mechanism might be related to the relief of endoplasmic reticulum stress.
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Objective:To evaluate the effect of repetitive transcranial magnetic stimulation (rTMS) on cognitive dysfunction after traumatic brain injury (TBI). Methods:Randomized controlled trials (RCTs) of rTMS for treating traumatic brain injury (TBI) patients with cognitive dysfunction were retrieved from the databases of PubMed, Embase, Web of Science, Cochrane Library, CNKI, Wanfang Database, VIP and SinoMed from the establishment of these databases to June, 2021. Two researchers independently screened the articles, extracted the data, and evaluated the quality. Meta-analysis was performed using RevMan 5.4. Results:A total of ten RCTs with 368 patients were included. Subgroup analysis showed that rTMS could improve the single cognitive function (SMD = 1.00, 95%CI 0.04 to 1.96, P = 0.04) but for overall cognitive function (SMD = 0.47, 95%CI -0.05 to 0.98, P = 0.08). rTMS was well tolerated, that the adverse reactions such as dizziness and mild headache were not significantly different from the control group (RR = 1.67, 95%CI 0.98 to 2.86, P = 0.06). Conclusions:It is still uncertain in the effectiveness of rTMS on cognitive dysfunction after TBI, but it is well tolerated.
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Hepatocellular carcinoma (HCC) is a prevalent and highly malignant cancer throughout the world. Effective treatment of this disease is impeded by the high rate of metastasis, recurrence, and chemoresistance. Recent studies have revealed the close relationship between the malignant phenotype of HCC and cancer stem cells (CSCs). Therefore, CSC-targeted therapy is considered a promising strategy to eradicate HCC. Traditional Chinese medicine (TCM) can be effective in preventing recurrence and metastasis of some advanced HCC. A growing amount of literature has discovered that extracts or compounds derived from TCM exert an anti-CSC effect. This review introduces some formulas and chemical compounds derived from TCMs that have been reported to inhibit CSCs of HCC; these TCM-related drugs may help to provide an alternative approach to help manage cancers, especially for HCC which has a great potential of metastasis, recurrence, and chemoresistance.
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Objective To investigate the molecular protective mechanisms of myocardial hypertrophic preconditioning by mitochondrial quantitative proteomics. Methods Fourteen C57BL6/ J male mice were randomly divided into sham group(n = 6) and cardiac hypertrophy preconditioning group(n = 8). The murine model of cardiac hypertrophy preconditioning was established by imposing transverse aortic constriction for 3 days and debanding the aorta for 4 days. Three mice from sham group and four mice from cardiac hypertrophy preconditioning group were randomly selected for proteomic analysis, and the remaining mice were used for functional and morphological experiments. The cardiac function was detected by echocardiography, and mechanical properties of cardiomyocytes were assessed using a SoftEdge Myocam. Cardiac morphology and mitochondrial ultrastructure were detected by pathological sections and transmission electron microscopy. The most significant mitochondrial proteins were screened by label-free quantitative proteomics and analyzed by bioinformatics analysis. Western blotting was used to verify the expression changes. Results Compared with the sham group, there were no significant changes in cardiac function and myocardial tissue morphology in the cardiac hypertrophy preconditioning group. However, electron microscopy analyses showed that the density of mitochondrial cristae increased in cardiac hypertrophy preconditioning group. Proteomic analysis screened 20 differentially expressed mitochondrial proteins. Bioinformatics analysis revealed that differentially expressed proteins were mainly related to mitochondrial ribosomal proteins. Western blotting results of key proteins were consistent with proteomic analysis. Conclusion Myocardial hypertrophic preconditioning can promote the energy metabolism of myocardial mitochondria, which may be related to the transcription, processing and transportation of mitochondrial oxidative phosphorylation complex mediated by mitochondrial ribosomal proteins.
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Objective:To observe the effect of Chaibei Zhixian decoction and peimine on Carbamazepine (CBZ) concentration, P-glycoprotein (P-gp) and multi drug resistance 1(MDR1) expression in the brain tissues of rats with refractory epilepsy, and to understand the contribution of Peimine in the compound prescription to treat the refractory epilepsy. Method:Epilepsy rat models were established by injecting kainic acid (KA) in the lateral ventricle. The successfully modeled rats were randomly divided into model group, CBZ group(0.12 g·kg-1),Chaibei Zhixian decoction+CBZ group(8.39 g·kg-1+0.12 g·kg-1), peimine+CBZ group(0.01 g·kg-1+0.12 g·kg-1) and sham operation group. After 60 days of intervention, the expression levels of P-glycoprotein (P-gp) and MDR1b mRNA in the brain cortex were detected by Western blot and quantitative real\|time fluorescence polymerase chain reaction(Real-time PCR),the contents of CBZ and 10,11-epoxidation of carbamazepine (CBZE) were measured by liquid chromatography-mass spectrometry (LC-MS). Result:Compared with sham group, the expression of P-gp/MDR1 in the cortex of model group was significantly increased (PPPPPPPConclusion:Chaibei Zhixian decoction and peimine may increase the content of CBZ and CBZE in the brain tissues in rats with intractable epilepsy by reducing the expression of MDR1/P-gp in the cortex.
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This prospective study was conducted to compare risk factors and pregnancy outcomes between women with complete placenta previa and those with incomplete placenta previa diagnosed in mid-pregnancy.The study was carried out from April 2014 to December 2015,during which 70 patients with complete previa and 113 with incomplete previa between 20+0 weeks and 25+6 weeks of gestation were included.Maternal demographics and pregnancy outcomes were compared between the two groups.Comparisons between categorical variables were tested by chi-squared test and those between continuous variables by Student t test.Resolution ofprevia occurred in 87.43％ of the studied women.The mean gestational age at resolution was 32.1±4.4 weeks.Incidence of maternal age ≥35 years and incidence of prior uterine operation ＞3 were high in women with complete previa (28.6％vs.8.8％,P=0.003;28.6％ vs.8.8％,P=0.003).Resolution of previa occurred less often in complete previa group (74.3％ vs.95.6％,P=0.001).Women with complete previa admitted earlier (37.3±2.0 weeks vs.38.1±1.4 weeks,P=0.011) and delivered earlier (37.7±1.2weeks vs.38.3±1.4 weeks,P=0.025).Maternal age ≥35 years and prior uterine operation ＞3 increase the risk of complete previa in mid-pregnancy.Placenta previa is more likely to persist in women with complete previa than those with incomplete previa diagnosed in midpregnancy.What is more,women with complete previa in mid-pregnancy deliversearlier.
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This prospective study was conducted to compare risk factors and pregnancy outcomes between women with complete placenta previa and those with incomplete placenta previa diagnosed in mid-pregnancy.The study was carried out from April 2014 to December 2015,during which 70 patients with complete previa and 113 with incomplete previa between 20+0 weeks and 25+6 weeks of gestation were included.Maternal demographics and pregnancy outcomes were compared between the two groups.Comparisons between categorical variables were tested by chi-squared test and those between continuous variables by Student t test.Resolution ofprevia occurred in 87.43％ of the studied women.The mean gestational age at resolution was 32.1±4.4 weeks.Incidence of maternal age ≥35 years and incidence of prior uterine operation ＞3 were high in women with complete previa (28.6％vs.8.8％,P=0.003;28.6％ vs.8.8％,P=0.003).Resolution of previa occurred less often in complete previa group (74.3％ vs.95.6％,P=0.001).Women with complete previa admitted earlier (37.3±2.0 weeks vs.38.1±1.4 weeks,P=0.011) and delivered earlier (37.7±1.2weeks vs.38.3±1.4 weeks,P=0.025).Maternal age ≥35 years and prior uterine operation ＞3 increase the risk of complete previa in mid-pregnancy.Placenta previa is more likely to persist in women with complete previa than those with incomplete previa diagnosed in midpregnancy.What is more,women with complete previa in mid-pregnancy deliversearlier.
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This prospective study was conducted to assess the rate of resolution of second trimester placenta previa in women with anterior placenta and posterior placenta,and that in women with and without previous cesarean section.In this study,placenta previa was defined as a placenta lying within 20 mm of the internal cervical os or overlapping it.We recruited 183 women diagnosed with previa between 20+0 weeks and 25+6 weeks.They were grouped according to their placenta location (anterior or posterior) and history of cesarean section.Comparative analysis was performed on demographic data,resolution rate of previa and pregnancy outcomes between anterior group and posterior group,and on those between cesarean section group and non-cesarean section group.Women with an anterior placenta tended to be advanced in parity (P=0.040) and have increased number of dilatation and curettage (P=0.044).The women in cesarean section group were significantly older (P=0.000) and had more parity (P=0.000),gravidity (P=0.000),and dilatation and curettage (P=0.048) than in non-cesarean section group.Resolution ofprevia at delivery occurred in 87.43％ women in this study.Women with a posterior placenta had a higher rate of resolution (P=0.030),while history of cesarean section made no difference.Gestational age at resolution was earlier in posterior group (P=0.002) and non-cesarean section group (P=0.008) than in anterior group and cesarean section group correspondingly.Placenta location and prior cesarean section did not influence obstetric outcomes and neonatal outcomes.This study indicates that it is more likely to have subsequent resolution of the previa when the placenta is posteriorly located for women who are diagnosed with placenta previa in the second trimester.
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Objective:To investigate the effect of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in modulat-ing the effects of oral squamous cell carcinoma (OSCC) invasion. Methods:Real-time polymerase chain reaction was employed to de-tect the expression of MALAT1 in samples of OSCC post-radical resection, normal oral mucosa samples, and oral squamous cell lines. MALAT1-siRNA was transfected into TSCCa human tongue squamous cell carcinoma cell lines. Cell proliferation was determined by methyl-thiazolyl-tetrazolium reduction assay. Cell migration and invasive ability were evaluated by scratch test and transwell assay. The expression of proteins that regulated invasion and apoptosis were examined using Western blot assay. Immunofluorescence assay was used to detect changes in epithelial-mesenchymal transition (EMT)-associated proteins in the cells. Tumor-bearing nude mouse models were established by subcutaneous implantation of TSCCa cells. Immunohistochemistry was used to detect up-regulation of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-2/9 (MMP-2/9). Results:MALAT1 expression was significantly higher in OSCC than in normal tissues (P<0.05). MALAT1 expression was inhibited by transfecting MALAT1-siRNA. After MALAT1 expres-sion was down-regulated in TSCCa cells, proliferation was inhibited and invasion was attenuated, showing significant differences com-pared with the cells transfected with scrambled siRNA and control cells (P<0.05). Expression of N-cadherin and MMP-2/9 were down-regulated in the cells after MALAT1 was knocked down. Tumor growth was significantly slower in the MALAT1-siRNA group than in the control groups. IHC indicated that PCNA and MMP-2/9 expression of tumor tissues were significantly inhibited in MALAT1-siR-NA group. Conclusion:MALAT1 is over-expressed in human OSCC. MALAT1 reduction can inhibit the proliferation and invasion of OSCC cells. Furthermore, MALAT1 may promote OSCC invasion and metastasis by modulating EMT.
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Objective To explore the influence of drug dosage, solvent and other main influencing factors on the successful establishment of alloxan-induced hyperglycemia mouse model and the effect on the stability of this model. Methods 160 6-8-week-old Kunming mice ofSPF grade, (male:female=1:1) were used in this study.The influences of different dosages of alloxan and solvent combinations on the successful establishment rate of the model, survival rate, body weight, fasting blood glucose, blood glucose area under curve, serum insulin level and their stabilities were dynamically observed for six weeks.Results By single intraperitoneal injection of 160 mg/kg bw alloxan ( pH 4.5 citrate sodium as solvent) , we were able to obtain a stable experimental hyperglycemic mouse model with higher levels of successful establishment rate (70%), survival rate (75%), fasting blood glucose (15-20 mmol/L), glucose area under the curve (55-65 mmol/L) and a lower but not loss of serum insulin levels (21 mIU/L).Conclusions In the present study we have carefully considered the influence of main factors such as drug dosages, solvent, etc., on the alloxan-induced experimental hyperglycemic mouse model, and successfully established this model after 6-week period observation of its stability.This model may provide a useful tool in the research of experimental diabetes and hypoglycemic functional studies.
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Objective: To study the chemical constituents from the leaves of Oplopanax horridus. Methods: The chemical constituents were isolated and purified by column chromatography on silica gel and Sephadex LH-20 gel columns, 1H-NMR and 13C-NMR were applied for the identification of chemical structure. Results: Ten compounds were isolated and identified as dammara-20,24-dien-3β-ol acetate (1), phytol (2), 16Z,19Z-pentacosadienoic acid (3), β-sitosterol (4), (3. S,8. S-falcarindiol (5), maltol (6), acankoreagenin (7), daucosterol (8), stigmasterol-3-. O-β-. D-glucopyranoside (9), and acankoreoside A (10). Conclusion: Compounds 1-3, 6, and 10 are isolated from this plant for the first time. Compounds 1-3 and 6 are isolated from the plants in genus Oplopanax Miq. for the first time. Moreover, Compounds 1, 3, and 6 are isolated from the plants in the family of Araliaceae for the first time.
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<p><b>OBJECTIVE</b>To sutdy the effect and mechanism of CXC chemokine receptor 4 (CXCR-4 )modulating oral squanmous cell carcinoma (OSCC) epithelial-mesenchymal transition (EMT)to mediate tumor lymphatic metastasis.</p><p><b>METHODS</b>Immunohistochemistry was used to detect the CXCR-4 and EMT related proteins among 60 OSCC samples with different lymph node metastasis status and the relationship with clinicopathological features and EMT related factors were analyzed. The small interfering RNA (siRNA) was applied to silence CXCR-4 in Tscca cell.Reverse transcription-PCR and Western blotting were used to observe the inhibitory effect. The wound healing assay, transwell invasion assay and flow cotometry were used to detect Tscca cell migration, invasion ability and apoptosis.</p><p><b>RESULTS</b>In lymph node metastasis group, the expression of CXCR-4, N-cadherin, Twist, Snail were higher than those in non-lymph node metastasis group. CXCR-4 showed a positive correlation with Twist, Snail, N-cadherin (correlation coefficient:0.300, 0.256, 0.333, P < 0.05), but negatively correlated with β-catenin (correlation coefficient:-0.497, P < 0.05). CXCR-4 silencing inhibited the migration and invasion of Tscca cell and induced apoptosis of the cell. Western blotting results indicated that N-cadherin, matrix metalloproteina-2/9 (MMP-2/9) was decreased (N-cadherin: F = 20.999, P = 0.002; MMP-2: F = 47.156, P = 0.000; MMP-9: F = 142.317, P = 0.000) while E-cadherin was up-regulated (F = 111.022, P = 0.000).</p><p><b>CONCLUSIONS</b>CXCR-4 might play a critical role in the metastasis of OSCC via modulating EMT.</p>
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Humans , Cadherins , Carcinoma, Squamous Cell , Pathology , Cell Movement , Epithelial-Mesenchymal Transition , Immunohistochemistry , In Vitro Techniques , Lymphatic Metastasis , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Mouth Neoplasms , Pathology , Receptors, CXCR4 , beta CateninABSTRACT
Oral squamous cell carcinoma (OSCC) is a common malignant tumor of the head and neck, and recurrence is an important prognostic factor in patients with OSCC. We explored the factors associated with recurrence of OSCC and analyzed the survival of patients after recurrence. Clinicopathologic and follow-up data of 275 patients with OSCC treated by surgery in the Cancer Institute and Hospital of Tianjin Medical University between 2002 and 2006 were analyzed. Recurrence factors were analyzed with Chi-square or Fisher's exact test and multivariate analysis. The prognosis of patients after recurrence was analyzed with the Kaplan-Meier method and log-rank test. The recurrence rate was 32.7%. The recurrence time ranged from 2 to 96 months, with a median of 14 months. Univariate analysis showed that T stage, degree of differentiation, pN stage, flap application, resection margin, and lymphovascular invasion were factors of recurrence (P < 0.05). Multivariate analysis showed that T stage, degree of differentiation, and pN stage were independent factors of recurrence (P < 0.001). The differences in gender, age, tumor site, region of lymph node metastasis, and perineural invasion between the recurrence and non-recurrence groups were not significant (P > 0.05). Kaplan-Meier and log-rank tests showed that the 2- and 5-year survival rates were significantly lower in the recurrence group than in non-recurrence group (67.6% vs. 88.0%, 31.8% vs. 79.9%, P < 0.001). Therefore, to improve prognosis, we recommend extended local excision, flap, radical neck dissection, and adjuvant chemoradiotherapy for patients more likely to undergo recurrence.