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Objective To investigate the effect of Ganlong capsule on axillary graft tumour in Balb/c nude mice.Methods BEL-7402,a human hepatocellular carcinoma sensitive strain,was inoculated into the armpit of nude mice for modeling.After successful modeling,mice were divided into model group,sorafenib group,CII-3 group,degreasing cream group,and Ganlong capsule high-dose,medium-dose and low-dose groups,and normal nude mice were included in blank group.All groups were given continuous administration for 14 days.The tumor inhibition rate,organ index,biochemical index,anoxic microenvironment related proteins and their mRNA expression levels were compared among all groups.Results All drug treatment groups could inhibit tumor growth,and the average tumor inhibition rate was the highest in Ganlong capsule medium-dose group.Compared with the model group,the heart index,kidney index,liver index and lung index of nude mice in all drug treatment groups were increased,and the spleen index were decreased,but most of the changes were little.Various biochemical indexes showed that Ganlong capsule was safe and did not cause obvious tissue damage.The mRNA expressions of hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR2)in sorafenib group and Ganlong capsule medium-dose group were significantly lower than those in model group(P<0.05).Immunohistochemical results showed that all drug treatment groups showed different degrees of inhibition on VEGFR2,HIF-1α,VEGF,Ki-67 and CD31 protein expressions,among which sorafenib group,Ganlong capsule medium-dose group and CII-3 group had stronger inhibitory effects.Conclusion Ganlong capsule can effectively inhibit the growth of axillary graft tumour in Balb/c nude mice,and down-regulate the expression level of hypoxic microenvironment related protein and their mRNA in the graft tumor.
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AIM To establish an UPLC-MS/MS method for the simultaneous content determination of chlorogenic acid,umbellifolactone,rutin,isochlorogenic acid A and isochlorogenic acid C in Artemisia sieversiana Ehrhart ex Willd.and Ajania tenuifolia(Jacquem.ex DC.)Tzvelev in Schischk.&Bobrov by HPLC,and compare the differences between them.METHODS The analysis of 50%methanol extract of both was performed on a 30℃thermostatic CAPCELL PAK-C18 column(100 mm×2.1 mm,1.7 μm),with the mobile phase comprising of acetonitrile-0.2%phosphoric acid water,flowing at 1 mL/min in a gradient elution manner,and the detection wavelengths were set at 290 nm.A t-test was used to compare the contents differences of five constituents in the two crude drugs.RESULTS Five constituents showed good linear relationships within their own ranges(r≥0.999 5),whose average recoveries were 96.03%-103.11%,with the RSDs of 0.51%-2.26%.The average contents of chlorogenic acid,umbellifolactone,rutin,isochlorogenic acid A and isochlorogenic acid C in A.sieversiana and A.tenuifolia were 3.61,0.71,1.46,8.74,1.44 mg/g and 2.60,0.26,0.79,6.31,1.22 mg/g,respectively.CONCLUSION The method is accurate,reliable and reproducible.There was no significant difference in the five chemical constituents between A.sieversiana and A.tenuifolia.
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In this study, untargeted metabolomics technology based on ultra-high-performance liquid chromatography-quadrupole/time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was used to analyze and identify the overall chemical components of Juniperri Caulis et Folium. Chemical markers for the identification of different Juniperri Caulis et Folium species were screened by integrated principal component analysis and partial least squares discriminant analysis. A total of 58 chemical components were detected and 46 of them were identified, including 26 flavonoids, 8 organic acids and their derivatives, 4 phenylpropanoids, 3 terpenoids, and 5 other components. Among them, methylsyringin and ekersenin were identified for the first time. In the positive ion mode, 12 markers were screened, and in the negative ion mode, 13 markers were screened for species identification. In summary, UPLC-Q-TOF-MS/MS metabonomics technology combined with chemometrics method can effectively reveal the chemical composition differences of different Juniperri Caulis et Folium species, and provide reference for its species identification and quality control.
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Minimal hepatic encephalopathy(MHE) is an early stage of hepatic encephalopathy with an insidious onset and a high rate of missed diagnosis in clinical practice, and it is of great importance to diagnose MHE as early as possible and provide effective clinical intervention. There are many diagnostic methods for MHE, among which psychometric hepatic encephalopathy score is the most commonly used method at present, but its wide application in clinical practice is limited by its complex and time-consuming operation, and therefore, it is urgent to find a simple, rapid, and effective clinical diagnostic method. Stroop test is a test for psychomotor speed and cognitive flexibility, and its value in the diagnosis of MHE has been verified in various countries including the United States and South Korea. This article introduces the development of Stroop test and its application in MHE, and the analysis shows that Stroop test based on mobile devices has a high sensitivity in the diagnosis of MHE and is simple, convenient, and feasible. It is hoped that this test can be widely used in the clinical work of MHE screening in China in the future.
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OBJECTIVE To esta blish the method for simultaneous determination of 25 components (such as berberine , magnoflorine and hydroxysafflor yellow A )in Bawei xiaobopi capsules. METHODS High-performance liquid chromatography- tandem triple quadrupole mass spectrometry (HPLC-QqQ-MS)method was adopted. The determination was performed on WondaSil C18-WR column with mobile phase consisted of 0.1% formic acid solution-methanol (gradient elution )at the flow rate of 0.5 mL/min. The column temperature was 25 ℃,and sample size was 5 μL. Electrospray ionization source was scanned in positive and negative ion mode at the same time ,with multiple reaction monitoring. The capillary voltage was 4 000 V(+)and 2 500 V(-). The drying gas flow rate was 11 L/min with the temperature of 300 ℃. The pressure was 15 psi. RESULTS Totally 25 components of Bawei xiaobopi capsules had good linear relationship within a certain range ,such as magnolflorine ,jatrorrhizine,berberine, palmatine,bufotenine,bufotenidine,piperine,glycyrrhizic acid ,ferulic acid ,ferulic acid 4-O-β-D-glucopyranoside,hydroxysafflor yellow A ,chlorogenic acid ,gallic acid ,chebulagic acid ,corilagin,ellagic acid ,liquiritigenin,liquiritin,rutin,quercetin, glycocholic acid ,cholic acid ,glycochenodeoxycholic acid ,glycodeoxycholic acid ,ursodeoxycholic acid (r≥0.999 0). The limits of quantitation were 0.62-554.50 ng/mL;the limits of detection were 0.18-166.30 ng/mL.RSDs of precision ,repeatability and stability(24 h)tests were all lower than 3.00%. The recovery rates were 80%-115%(all RSDs lower than 3.00%,n=6). The contents of above 25 components were 16.94-20.82,3.78-5.17,9.11-11.43,0.24-0.30,0.20-0.39,0.74-1.16,0.79-0.89,3.26-3.35, 0.48-0.66,11.96-13.35,2.30-3.12,0.19-0.21,6.07-8.83,10.42-10.48,1.43-1.64,4.17-4.76,0.14-0.15,0.46-0.52,0.04,0.01, 0.59-0.63,0.20-0.23,0.02,0.15-0.16,0.01 mg/g,respectively. CONCLUSIONS Established method is simple ,sensitive and stable,and can be used for content determination of 25 components in Bawei xiaobopi capsules simultaneously.
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Objective:To study the effect of crizotinib on acute radiation-induced lung injury in mice and its possible mechanism.Methods:A total of 72 mice were randomly divided into 4 groups by the random number table method: healthy control group (NC group, n=12), crizotinib-only group (CRZ group, n=12), radiotherapy-only group (RT group, n=24), and radiotherapy pluscrizotinib group (RT+ CRZ group, n=24). The whole lungs were exposed to a single dose of 12 Gy X-rays. Lung tissue and bronchoalveolar lavage fluid (BALF) were obtained at 1, 2, 4, and 8 weeks after radiotherapy. The total number of nucleated cells was counted under a light microscope, and the total protein content of BALF was detected by bicinchoninic acid (BCA) protein assay. The pathological changes of lung tissue were observed by HE staining and MASSON staining. The expressions of TGF-β1 and ICAM-1 mRNA in lung tissue were detected by real-time quantitative polymerase chain reaction (qPCR), the locations and expressions of MPO and ICAM-1 proteins were observed by immunohistochemical staining, and the expressions of TGF-β1, Smad3, p-Smad3 and ICAM-1 proteins in lung tissue were detected by Western blot. Results:At different time points after irradiation, the pathological manifestations such as inflammation and exudation of lung tissue in the RT+ CRZ group were significantly increased, and the total number of cells and protein content in BALF was higher than that of the other three groups, compared with RT group, the difference was statistically significant at 4 week ( t=-5.031, -2.814, P<0.05). Compared with RT group, the expressions of ICAM-1 and TGF-β1 mRNA in lung tissue of the RT+ CRZ group were significantly increased, while the expression of TGF-β1 increased significantly at 1, 4 and 8 weeks after irradiation ( t=-2.687, -7.032, -5.221, P<0.05), and the expression of ICAM-1 increased significantly at 2 and 4 weeks after irradiation ( t=-4.819, -6.057, P<0.05). The expressions of these two gradually increased from 1 to 4 weeks and peaked in 4 weeks, then decreased at 8 weeks. At the same time, the trend of the expression of MPO mRNA was consistent with ICAM-1 and TGF-β1. At 4 week, there was no difference in the expression of Smad3 protein in these four groups ( P>0.05). The expressions of TGF-β1, p-Smad3, ICAM-1 and p-Smad3/Smad3 proteins of the RT+ CRZ group were all higher than those of the other three groups ( F=14.74, 10.03, 35.29, 22.94, P<0.05). Conclusions:Crizotinib combined with radiotherapy can aggravate acute radiation-induced lung injury, which may due to the increase of ICAM-1 expression by up-regulating the TGF-β1 signaling pathway.
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Objective:To investigate the effect and mechanism of PAE<sub>2</sub>, a polypeptide of <italic>Periplaneta americana, </italic>in reversing multidrug resistance (MDR) for liver cancer <italic>in vivo</italic>. Method:Balb/c-nude mice were inoculated with HepG2 and HepG2/ADM cells under the armpits to establish animal models of liver cancer sensitive strains and animal models of MDR respectively. After successful modeling, the nude mice were randomly divided into normal group, HepG2 model group, HepG2/ADM model group, sorafenib group (positive drug control group, <italic>ig</italic> 30 mg·kg<sup>-1</sup>), HepG2/ADM+PAE<sub>2</sub> (<italic>iv</italic>) low, medium and high dose groups (50, 100, 200 mg·kg<sup>-1</sup>), HepG2/ADM+PAE<sub>2</sub> (<italic>ig</italic>) low, medium, and high dose groups (50, 100, 200 mg·kg<sup>-1</sup>), skim cream group (<italic>ig</italic> 200 mg·kg<sup>-1</sup>), and CⅡ-3 group (<italic>ig</italic> 200 mg·kg<sup>-1</sup>), all of which received corresponding drug treatment. The body weight and tumor volume of nude mice were measured and recorded every 2 days. The next day after the last administration, tumor tissues of nude mice were taken to record the tumor weight. The effect of <italic>P. americana </italic>polypeptide PAE<sub>2</sub> on permeability-glycoprotein(P-gp), lung resistance protein(LRP) , breast cancer resistance protein(BCRP), protein kinase C(PKC), glutathione S-transferase-π(GST-π), topo-isomerase typeⅡ(ToPoⅡ), multidurg resistance gene 1(MDR1)<sub> </sub>and Multidrug resistance-associated proteins(MRP1) of the protein level and gene level expression in tumor tissues were determined by immunohistochemistry (IHC) and real-time quantitative polymerase chain reaction (Real-time PCR). In addition, both oral and intravenous administration groups were set up at the same time for preliminary study on the basic pharmacokinetic characteristics of <italic>P. americana </italic>polypeptide PAE<sub>2</sub>. Result:After the successful modeling, the body weight of the nude mice was significantly lower than that in the normal mice(<italic>P</italic><0.05). After treatment with corresponding drugs, the body weight increased to a certain extent, but it was still not as good as the normal nude mice. In <italic>iv</italic> administration, the medium-dose <italic>P. americana </italic>polypeptide PAE<sub>2</sub> showed the best anti-tumor effect as compared with the model group (<italic>P</italic><0.05), while in oral administration, the anti-effect increased with the increase of the dose, so the high-dose group showed the best effect (<italic>P</italic><0.05). Preliminary crude extract CII-3 had no obvious anti-tumor effect, and skim cream showed a certain anti-tumor effect (<italic>P</italic><0.05). <italic>P. americana </italic>polypeptide PAE<sub>2</sub> had certain effects on MDR related proteins and enzymes<italic> in vivo</italic>, mainly by inhibiting the expression of LRP and BCRP in tumor tissues and affecting the expression of these related proteins and genes to different degrees to inhibit intracellular drugs outflow, thereby promoting tumor apoptosis, and the effect was superior to that of the <italic>P. americana</italic> crude extract CⅡ-3 and skim cream. Conclusion:<italic>P. americana</italic> polypeptide PAE<sub>2</sub> may reduce the drug efflux, promote intracellular drug accumulation and apoptosis by affecting the expression of related proteins and enzymes that mediate multidrug resistance, thereby exerting a reverse effect on HepG2/ADM cells Balb/c MDR in nude mice.
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Objective:To investigate the construction and application of telemedicine system in Tibet Autonomous Region.Methods:From June to July 2020, medical institutions at all levels in Tibet Autonomous Region were visited. Relevant information was collected through discussion, questionnaires and field visits, and the data of effective questionnaires were statistically analyzed.Results:A total of 125 medical institutions were visited, covering 7 regions of Tibet Autonomous Region, of which 74 medical institutions were able to realize audio-video two-way communication with the telemedicine platform of the PLA General Hospital. Seventy-three valid questionnaires were collected, and 64(88%)hospitals had software or hardware audio and video terminals. Twenty-five hospitals had never carried out remote consultation, accounting for 34%. The annual remote consultation amount of 35 hospitals was less than 10 cases, and that of 11 hospitals was 11-100 cases. Only 2 hospitals had carried out remote ECG and remote ultrasound diagnosis, with an annual consultation volume of more than 100 cases.Conclusions:The hospitals at or above the county level in Tibet Autonomous Region have established telemedicine system, but there are few remote services, the system idle rate is high, and the distance education resources are not matched.
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Objective:To evaluate the 5-year survival outcome of patients with unresectable locally advanced non-small cell lung cancer (NSCLC) treated with Endostar in combination with platinum-based concurrent chemoradiotherapy.Methods:From March 2009 to June 2015, 115 patients with the unresectable locally advanced NSCLC from two prospective studies[Clinical trials 2009-2012(ClinicalTrials.gov NCT01894) and 2012-2015(ClinicalTrials.gov, NCT01733589)] were treated with Endostar in combination with platinum-based concurrent chemoradiotherapy. A total dose of 60-66 Gy was delivered in 30-33 fractions. Endostar was given 1 week prior to the beginning of radiotherapy, and repeated fortnightly during the concurrent chemoradiotherapy. After long-term follow up, survival outcome was evaluated in 104 patients treated with radiation dose of ≥60 Gy. Kaplan-Meier method was used for survival analysis. Univariate survival analysis was performed using the log-rank test.Results:Of 104 eligible patients, 60.6% of them had squamous carcinoma and 65.4% were classified in stage Ⅲ B. All the patients received ≥2 cycles of Endostar and 93.3% of them received 4 cycles of Endostar. The median follow-up time was 68.3 months. The median overall survival (OS) and median progression-free survival (PFS) were 31.3 and 13.9 months, respectively. The 3-year and 5-year OS were 45.6% and 35.7%, respectively. The 3-year and 5-year PFS were 27.1% and 24.9%, respectively. Univariate analysis indicated that sex, ECOG, pathological type, clinical stage, radiotherapy technique, chemotherapy regimen, chemotherapy cycle and cycle of Endostar use were not associated with OS. Late radiation injury occurred in 14.4% of patients, and no grade 4-5 late injury was observed. Conclusion:Patients with unresectable locally advanced NSCLC treated with Endostar fortnightly in combination with platinum-based concurrent chemoradiotherapy achieve better OS than historical data with tolerable toxicities.
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Objective:To investigate the current status of prevention and treatment of esophagogastric variceal bleeding (EVB) in cirrhotic portal hypertension patients in Ningxia region.Methods:The retrospective and descriptive study was conducted. The clinical data of 820 cirrhotic portal hypertension patients who were admitted to 21 medical centers in Niangxia region from January 2018 to December 2020 were collected, including 85 cases in Ningxia Hui Autonomous Region People′s Hospital, 73 cases in the Fifth People′s Hospital of Ningxia Hui Autonomous Region, 59 cases in the Wuzhong People′s Hospital, 52 cases in the Qingtongxia People′s Hospital, 50 cases in the Guyuan People′s Hospital, 47 cases in the Yuanzhou District People′s Hospital of Guyuan City, 47 cases in the Yinchuan Second People′s Hospital, 40 cases in the General Hospital of Ningxia Medical University, 40 cases in the Tongxin People′s Hospital, 35 cases in the Yinchuan First People′s Hospital, 34 cases in the Third People′s Hospital of Ningxia Hui Autonomous Region, 32 cases in the Zhongwei People′s Hospital, 30 cases in the Lingwu People′s Hospital, 30 cases in the Wuzhong New District Hospital, 30 cases in the Yanchi People′s Hospital, 29 cases in the Ningxia Hui Autonomous Region Academy of Traditional Chinese Medicine, 28 cases in the Shizuishan Second People′s Hospital, 25 cases in the Shizuishan First People′s Hospital, 21 cases in the Haiyuan People′s Hospital, 20 cases in the Pengyang People′s Hospital, 13 cases in the Longde People′s Hospital. There were 538 males and 282 females, aged (56±13)years. Observation indicators: (1) clinical charac-teristics of cirrhotic portal hypertension patients; (2) overall prevention and treatment of EVB in cirrhotic portal hypertension patients; (3) prevention and treatment of EVB in cirrhotic portal hypertension patients from different grade hospitals. Measurement data with normal distribution were represented as Mean± SD. Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test. Results:(1) Clinical characteristics of cirrhotic portal hypertension patients: of 820 cirrhotic portal hypertension patients, 271 cases were in compensated stage and 549 cases were in decompensated stage. Of the 271 cases in compensated stage, there were 183 maels and 88 females, aged (53±12)years. There were 185 Han people, 85 Hui people and 1 case of other ethic group. The etiological data of liver cirrhosis showed 211 cases of viral hepatitis B, 4 cases of alcoholic liver disease, 8 cases of viral hepatitis C, and 48 cases of other etiology. There were 235 cases of Child-Pugh grade A and 36 cases lack of data. Of the 549 cases in decompensated stage, there were 355 males and 194 females, aged (57±14) years. There were 373 Han people, 174 Hui people and 2 cases of other ethic group. The etiological data of liver cirrhosis showed 392 cases of viral hepatitis B, 33 cases of alcoholic liver disease, 10 cases of viral hepatitis C, and 114 cases of other etiology. There were 80 cases of Child-Pugh grade A, 289 cases of grade B, 170 cases of grade C and 10 cases lack of data. (2) Overall prevention and treatment of EVB in cirrhotic portal hypertension patients: of 271 patients in compensated stage, 38 cases received non-selective β-blocker (NSBB) therapy, 16 cases received endoscopic treatment, 6 cases received interventional therapy. Of 549 patients in decompensated stage, 68 cases received NSBB therapy, 46 cases received endoscopic treatment, 28 cases received interventional therapy. (3) Prevention and treatment of EVB in cirrhotic portal hypertension patients from different grade hospitals: of 271 patients in compensated stage, 181 cases came from tertiary hospitals, of which 28 cases received NSBB therapy, 15 cases received endoscopic treatment, 6 cases received interventional therapy. Ninety cases came from secondary hospitals, of which 10 cases received NSBB therapy, 1 cases received endoscopic treatment. There was no significant difference in NSBB for prevention of EVB between tertiary and secondary hospitals ( χ2=0.947, P>0.05), while there was a significant difference in endoscopic treatment for prevention of EVB between tertiary and secondary hospitals ( χ2=5.572, P<0.05). Of 549 patients in decompensated stage, 309 cases came from tertiary hospitals, of which 22 cases received NSBB therapy, 29 cases received endoscopic treatment, 22 cases received interventional therapy. Two hundreds and fourty cases came from secondary hospitals, of which 46 cases received NSBB therapy, 17 cases received endoscopic treatment, 6 cases received interven-tional therapy. There were significant differences in NSBB and interventional therapy for prevention of EVB between tertiary and secondary hospitals ( χ2=18.065, 5.956, P<0.05). Conclusions:The proportion of receiving EUB prevention in cirrhotic portal hypertension in Ningxia is relatively low. For patients with compensated liver cirrhosis, the proportion of NSBB therapy and endoscopic treatment in the secondary hospitals was lower than that in tertiary hospitals. For patients with decompensated liver cirrhosis, the proportion of interventional treatment in secondary hospitals is lower than that of tertiary hospitals, but the proportion of NSBB in secondary hospitals taking is higher than that of tertiary hospitals.
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Objective:To quickly analyze and identify the differential chemical compositions of Aurantii Fructus Immaturus before and after stir-frying with bran and chemical compositions of wheat bran after processing by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MSE) combined with UNIFI database screening method. Method:ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 µm) was used for chromatographic separation with 0.1% formic acid solution (A)-acetonitrile (B) as the mobile phase for gradient elution (0-11 min, 98%-70%B; 11-15 min, 70%-55%B; 15-16 min, 55%-35%B; 16-20 min, 35%-5%B; 20-20.5 min, 5%-98%B; 20.5-22 min, 98%B) at the flow rate of 0.3 mL·min-1 and the injection volume of 2 µL. The analytes were determined in positive ion mode with electrospray ionization (ESI) and data collection range of m/z 50-1 500. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to find the component differences between raw and processed products of Aurantii Fructus Immaturus, and the chemical compositions of wheat bran after processing were determined. Result:There were 64 compounds in raw products, 58 compounds in bran-fried products, and 18 compounds in wheat bran.There were 19 different components between raw and processed products of Aurantii Fructus Immaturus, mainly volatile oil, flavonoids, phenolic acid, coumarins and saponins. Conclusion:Based on the analysis of these different components before and after stir-frying with bran and the chemical compositions carried by wheat bran, the stir-frying with bran can alleviate the intensity of Aurantii Fructus Immaturus, which proves the necessity of stir-frying with bran for the processing technology of this herb, and provides a comprehensive experimental basis for research on processing mechanism of Aurantii Fructus Immaturus.
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Objective::To deeply understand important scientific problems and key technical problems in the cultivation of Chuanxiong, the Chuanxiong cultivation situation of 247 households in 23 towns of 5 counties in Sichuan province were investigated, providing basic data for the standardized cultivation of Chuanxiong. Method::On the basis of reported literatures, a questionnaire survey was conducted on the farmers of cultivating Chuanxiong in main producing areas, field measurements were also preformed, and the cultivation status of Chuanxiong was comprehensively analyzed. Result::The proportion of plain-breeding mainly existed in Pengzhou and Shifang, where per farmer had a small scale of planting areas, was 65%, while the proportion of mountain-breeding mainly existed in Meishan, Qionglai and Dujiangyan, where per proprietor had a large scale of planting areas, was 92%. The planting density and yield of Chuanxiong varied greatly among individuals. The growth period of Chuanxiong in Meishan and Qionglai is about 30 days shorter than that in Pengzhou, Shifang and Dujiangyan. About half of farmers applied base fertilizer and top dressing in spring. The proportion of NPK compound fertilizer input was high (>90%). Chuangxiong has a few diseases and insect pests, the incidence of which in new production areas was obviously lower than that in old production areas. The application of herbicide was not standardized. 52% farmers used Chu cao ling No.1 which was made by agricultural material store owners. The labor cost accounted for the highest proportion (37%) in Chuanxiong cultivation. Conclusion::Now in Sichuan clonal propagated patterns of Chuanxiong were both mountain-breeding and plain-breeding. It is suggested to make clear the differences between mountain-breeding nodes and plain-breeding nodes, and breed high qualities of Chuanxiong nodes. The mechanization research and production of Chuanxiong should be promoted, realizing timely planting and harvesting. Farmers should be guided to increase the application of organic fertilizer, and meanwhile standardize the application of agrochemicals.
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@#AIM: To observe the effects of butterflybush flower eye drops at different concentrations on expression of inflammatory crtokines IL-1β, Mucin 5AC(MUC5AC)and P38MAPK in castrated male rabbits, and to explore the therapeutic effect of that drops on dry eyes. <p>METHODS: Thirty-six male rabbits were randomly divided into blank group(A), model group(B), low concentrations butterflybush flower eye drops group(C, 1mg/mL), the medium concentrations drops group(D, 1.5mg/mL), the high concentrations drops group(E, 3mg/mL), and testosterone group(F). In addition to group A, the testes and epididymis were removed from each group to establish a dry eye animal model. After successful modeling, groups A and B remain unchanged. Groups C, D, and E were given different concentrations of butterflybush flower eye drops, 3 times/d. In group F, testosterone propionate was injected into the muscles of the thigh at a dose of 0.5mL/kg once every 3d. Fluorescein staining, Schirmer I test(SⅠt)and tear film break time(BUT)were measured under general anesthesia in each group, eatment. After 4wk of treatment, the rabbits were sacrificed and the conjunctival tissues of the eyes were taken. The expression of IL-1β, mucin 5AC and P38MAPK in the conjunctiva was detected by immunohistochemical staining.<p>RESULTS: Among low concentrations butterflybush flower eye drops group, the medium concentrations drops group and the high concentrations drops group, the SⅠt value was significantly higher than that of model group, and BUT was significantly longer than model group. The positive staining of corneal fluorescein was significantly improved compared with model group, which was statistically significant(<i>P</i><0.01). Among IL-1β and P38MAPK in the conjunctiva of high concentrations butterflybush flower eye drops group, the medium concentrations drops group and the low concentrations drops group, the positive expressions were lower than those in model group, and the expression of MUC5AC was higher than that in group model group(<i>P</i><0.01). In addition, the high concentrations drops group was superior to the low and the medium concentrations drops group.<p>CONCLUSION: Butterflybush flower eye drops have androgen-like effect. For castrated dry eyes of male rabbits, they can down-regulate the expression of IL-1beta and P38MAPK in dry conjunctival tissue and increase the expression of MUC5AC, thus reducing inflammation infiltration in dry conjunctival tissue and maintaining tear film stability, but their effect is weaker than that of androgen. To the treatment of dry eyes, the middle and high concentration groups of the drops had stronger effects than the low one, and the high concentration group was better than the medium one.
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OBJECTIVE@#To compare the bonding durability of three different self-etch adhesives to primary enamel and dentin, and to investigate the effect of mild salivary contamination followed by air drying on the bonding durability.@*METHODS@#Two hundred and forty enamel specimens were divided randomly into 16 groups (n=15/group)according to the adhesive system [self-etch adhesives: Clearfil SE Bond(SE), AdperTM Easy One (EO), Scotchbond Universal (SBU); total-etch adhesive: AdperTM Single Bond Plus(SL)], contamination status (non-contaminated vs. salivary-contaminated) and storage condition (stored in distilled water for 24 h vs. aging mode 5 000 thermal cycles in 5 ℃ and 55 ℃). Two hundred and forty dentin specimens were assigned in the same way. Shear bond strength for 12 specimens in each group were measured. The adhesive interface for the residual specimens in each group was observed by scanning electron microscopy(SEM). Data were analyzed by three-way analysis of variance and Tukey test(P < 0.05).@*RESULTS@#For primary enamel, total-etch adhesive showed higher initial shear bond strength values (28.92±1.83) MPa and shear bond strength values (27.27±3.03) MPa after thermal cycles compared with the other groups, and the difference between the groups was statistically significant (P < 0.01). Shear bond strength values of EO decreased significantly in salivary-treated groups, regardless of storage conditions, and the difference was statistically significant (P < 0.01). For primary dentin, shear bond strength values of EO decreased significantly in salivary-treated groups after 24 h (P < 0.01). After 5 000 thermal cycles, total-etch adhesive showed significantly lower shear bond strength values (14.31±1.97) MPa compared with the other groups, and the difference between the groups was statistically significant (P < 0.01), and shear bond strength values of EO were significantly lower than those in SE and SBU groups (P < 0.01), regardless of contamination status.@*CONCLUSION@#Total-etch adhesive SL has better bonding durability to primary enamel. SE and SBU have better bonding durability to primary dentin and have a certain resistance to salivary contamination, while the bonding performance of EO is compromised greatly by mild salivary contamination followed by air drying.
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Condicionamento Ácido do Dente , Adesivos , Colagem Dentária , Dentina , Adesivos Dentinários , Teste de Materiais , Cimentos de Resina , Resistência ao Cisalhamento , Dente DecíduoRESUMO
OBJECTIVE:To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3(shorted for “degreasing cream ”and“CⅡ-3”)reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS :MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib (positive control ),degreasing cream and C Ⅱ-3 on HepG2/ADM cells ,then IC 20 was calculated. The experiment was divided into sensitivity drug ,drug-resistance group ,sorafenib group,degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ,and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ,and other 3 groups were treated with relevant medicine (IC20 as drug concentration ). The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp (expression produce of MDR1 gene),LRP,BCRP] and that related to enzyme-mediated multidrug resistance pathway (GST-π and Topo Ⅱ). RESULTS :The IC 20 of degreasing cream ,CⅡ-3 and sorafenib were (2.40±0.16), (200.44±27.52),(18.00±1.82)μg/mL,respectively. Compared with sensitivity group ,the protein expressions of Bcl- 2,P-gp, LRP,BCRP and Topo Ⅱ,the mRNA expressions of MDR 1, LRP,BCRP and GST-π were increased significantly in drug resistance group (P<0.05 or P<0.01). Compared with @qq.com drug-resistance group ,the mRNA and protein expression of MDR1 mRNA and LRP ,BCRP,GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group(P< 0.05 or P<0.01);the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased (P<0.01), while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group (P<0.05). CONCLUSIONS:Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux , promoting cell apoptosis ,reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.
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OBJECTIVE@#To study the regulatory mechanism of MS275, a histone deacetylase inhibitor, on the p38 MAPK signaling pathway in rats with convulsion in the developmental stage.@*METHODS@#Thirty-two male rats were randomly divided into four groups: control, pentylenetetrazol (PTZ), PTZ+3 mg/kg MS275, and PTZ+6 mg/kg MS275 (n=8 each). A rat model of convulsion in the developmental stage was prepared by an intraperitoneal injection of PTZ. The rats in the control group were given an injection of normal saline alone. MS275 was given by an intraperitoneal injection at 2 hours before PTZ injection. At 24 hours after successful modeling, 6 rats were taken from each group. Western blot and qRT-PCR were used to measure the protein and mRNA expression of p38, MK2, cAMP response element-binding protein (CREB), and interleukin-6 (IL-6) in the hippocampus. Hematoxylin-eosin (HE) staining was used to observe brain pathological changes. Western blot was used to measure the expression of CD11b as a marker for the activation of microglial cells.@*RESULTS@#Compared with the control group, the PTZ group had significant increases in the mRNA and protein expression of p38, MK2, CREB, and IL-6 (P<0.05). MS275 significantly inhibited the mRNA and protein expression of the above markers in the rats with convulsion in the developmental stage (P<0.05), and 6 mg/kg MS275 had a significantly better inhibitory effect on the mRNA and protein expression of IL-6 and CREB than 3 mg/kg MS275 (P<0.05). HE staining showed that the PTZ group had marked neuron apoptosis, cellular edema, and inflammatory cell infiltration, while MS275 intervention alleviated neuron apoptosis and cellular edema and reduced inflammatory cell infiltration in the rats with convulsion. The PTZ group had a significant increase in the activation of microglial cells, while MS275 significantly inhibited the activation of microglial cells in the rats with convulsion (P<0.05); 6 mg/kg MS275 had a significantly better inhibitory effect than 3 mg/kg MS275 (P<0.05).@*CONCLUSIONS@#In rats with convulsion in the developmental stage, the histone deacetylase inhibitor MS275 can inhibit the p38 MAPK signaling pathway, the apoptosis of hippocampal neurons, and the activation of microglial cells and thus reduce inflammatory response and convulsion-induced brain injury in a dose-dependent manner.
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Animais , Masculino , Ratos , Pentilenotetrazol , Ratos Sprague-Dawley , Convulsões , Transdução de Sinais , Proteínas Quinases p38 Ativadas por MitógenoRESUMO
OBJECTIVE@#To assess the changes in the coagulation profiles of patients with chronic kidney disease (CKD) using thromboelastography (TEG) and identify the risk factors of hypercoagulation in CKD patients.@*METHODS@#A total of 128 patients with CKD admitted in Hunan Provincial People's Hospital between August, 2018 and May, 2019 were recruited. The results of conventional coagulation test and TEG were compared between patients with CKD and 21 healthy control adults. The patients with CKD were divided into hypercoagulation group with a maximum amplitude (MA) > 68 mm (=66) and non-hypercoagulation group (MA≤68 mm, =62). The laboratory indicators were compared between the groups, and the factors affecting the hypercoagulable state in patients with CKD were analyzed.@*RESULTS@#The levels of fibrinogen and D-Dimer increased significantly in patients with CKD at different stages as compared with the control subjects ( < 0.05). In the patients with CKD, the reaction time and K time decreased while MA, α-angle and coagulation index increased significantly in patients in stage 3-4 and those in stage 5 either with or without hemodialysis compared with the control group ( < 0.05). The estimated glomerular filtration rate (eGFR), percentage of patients with diabetes mellitus, history of stroke, percentage of neutrophils, neutrophil-lymphocyte ratio, red blood cell count, hemoglobin levels, platelet count, serum creatinine, serum cystatin-C, serum albumin, and lipoprotein (a) all differed significantly between hypercoagulation group and non-hypercoagulation group ( < 0.05). The eGFR, platelet count and hemoglobin levels were identified as independent factors affecting hypercoagulability in patients with CKD ( < 0.05).@*CONCLUSIONS@#s The hypercoagulable state of patients with CKD worsens gradually with the disease progression, and eGFR, platelet count and hemoglobin levels are all risk factors for the hypercoagulable state in patients with CKD.
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Humanos , Coagulação Sanguínea , Insuficiência Renal Crônica , Fatores de Risco , Tromboelastografia , TrombofiliaRESUMO
OBJECTIVES@#To detemine preventive effects of compound formula Rhizoma Coptidis and Atractylodes on mice with gastric-ulcer.@*METHODS@#The mice were randomly divided into a normal group, a gastric ulcer group, a ranitidine positive drug group, a Rhizoma Coptidis group, an Atractylodes group, and a Rhizoma Coptidis plus Atractylodes group (the ratios of Coptidis to Atractylodes were 9꞉1, 8꞉2, 7꞉3, 6꞉4, 5꞉5, or 4꞉6, respectively). Gastric ulcer models were established by intragastric administration of anhydrous ethanol after 6 days of preventive infusion. The mice were killed 6 days after the treatments. The whole stomach was opened to observe gross morphology of gastric mucosa. The pathological changes of gastric tissue were observed under microscope, and serum samples were collected to detect the contents of superoxide dimutase (SOD), malondialdehyde (MDA), NO, and endothelin-1 (ET-1).@*RESULTS@#The Rhizoma Coptidis and Atractylodes decoction significantly decreased ulcer area (<0.001), and the effects of compound formula are better than those of Coptidis and Atractylodes alone (<0.05, <0.01, or <0.001). The anti-ulcer effect of compound formula (Coptidis꞉Atractylodes=6꞉4) was the best one, and the anti-gastric ulcer effect of the high-dose group was significantly better than that of the ranitidine-positive group (<0.001). The ranitidine positive drug group, the high-dose group of Rhizoma Coptidis, the high-dose group of Atractylodes, and the high-dose group of Rhizoma Coptidis-Atractylodes (6꞉4) significantly reduced MDA, ET-1 (<0.01 or <0.001), and significantly increased SOD, NO in serum (<0.01 or <0.001).@*CONCLUSIONS@#Rhizoma Coptidis and Atractylodes decoction exerts the effect on preventing ethanol-induced gastric ulcer in mice in a ratio-dependent and dose-dependent manner. The mechanism might be related to anti-oxidation and relaxion of blood vessels. The combination of the two drugs shows a synergistic effect.
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Animais , Camundongos , Atractylodes , Medicamentos de Ervas Chinesas , Mucosa Gástrica , Úlcera GástricaRESUMO
Objective: To investigate the effect of the Periplaneta Americana polypeptide on the angiogenesis. Method:Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cell scratch assay were used to observe effect of different concentration (6.25,12.5,25,50,100 mg·L-1) of the Periplaneta Americana polypeptide, CⅡ-3 and skimmed cream on the proliferation and migration of human umbilical vein endothelial cells (HUVECs), and a normal group and a thalidomide group were also established in this study. The tubule formation assay was used to detect the effect of different concentration (25, 50, 100 mg·L-1) of the Periplaneta Americana extracts on the formation of tubules in HUVECs cells. The adhesion between HepG2 cells and HUVECs cells was observed by cell adhesion assay. The expression of vascular endothelial growth factor (VEGF) proteins in HUVECs was detected by immunocytochemical staining and enzyme linked immunosorbent assay (ELISA). Result:MTT results showed that the Periplaneta Americana polypeptide could inhibit the proliferation of HUVECs in a dose-dependent manner (PPPPPPPPPConclusion:The Periplaneta Americana polypeptide can inhibit the invasion, metastasis and tube formation of HUVECs, and down-regulate the expression of VEGF in HUVECs. The effect of Periplaneta Americana polypeptide is better than CⅡ-3 and skimmed cream, and the among the polypeptide, the effect of PAP-2 is superior to the other two.
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OBJECTIVE@#To evaluate the practical value of nearinfrared light transillumination (NILT, DIAGNOcam) on detection of incipient proximal caries in primary molars.@*METHODS@#In this study, 4-9-year-old children with suspicious proximal lesions in primary molars were recruited. The target teeth were examined with clinical examination, bitewing radiograph and DIAGNOcam. And the caries indexes were rated as 0 (no caries), 1 (caries reaching outer 1/2 enamel), 2 (caries reaching inner 1/2 enamel), and 3 (dentine caries). Those primary molars with at least one of the adjacent molars according with the criteria of invasive treatment, i.e. the teeth were diagnosed as caries reaching inner 1/2 enamel or dentine with at least two detection methods, were included. The target teeth with caries index ≥2 accepted the invasive treatment. And the caries status of the adjacent teeth was observed under direct vision and those teeth of caries index ≥2 would also accept the invasive treatment. The relationship of lesions to enamel-dentine junction (EDJ) were recorded and used as reference standard to compare the sensitivity, specificity and accuracy of those three methods in detecting the incipient proximal caries in primary molars.@*RESULTS@#In this study, 36 children with 104 suspicious proximal lesions in primary molars were recruited, of which 5 were diagnosed as no caries, 20 were diagnosed as superficial enamel caries, and 79 accepted invasive treatment, of which 22 were diagnosed as EDJ caries, and 57 as dentine caries. The sensitivity of clinical examination, bitewing radiograph, and DIAGNOcam were 68.69%, 81.82%, 78.79%, respectively. The specificity were 100.00%, 100.00%, 80.00%, and the accuracy were 75.96%, 85.65%, 78.85%, respectively. As for superficial enamel caries, the sensitivity of clinical examination was lowest, only 10%, while DIAGNOcam was 60%, higher than 40% of bitewing radiograph. As for EDJ caries, the sensitivity of bitewing radiograph was highest as 86.36%, followed by DIAGNOcam 72.73% and clinical examination 68.18%. As for dentine caries, the sensitivity of the three detection methods were higher than 85%, of which bitewing radiograph was the highest as 94.74%.@*CONCLUSION@#The accuracy of DIAGNOcam in detecting incipient proximal caries in primary molars was comparable to that of bitewing radiograph. It was safe, and convenient with no exposure to radiation, which was applicable to be used in pediatric dentistry.