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Objective: To investigate the effects of Zhongfeng capsule on the autophagy-related proteins expression in rats with cerebral ischemia/reperfusion injury (CI/ RI), and to explore its neural protection mechanisms of the decoction. Methods: Rat middle cerebral artery ischemia/reperfusion injury model (ischemia for 2 h, reperfusion for 24 h) was prepared by the improved line plug method. Sixty male SD rats were randomly divided into sham operation group, model group, butylphthalide group(0.054 g/kg), Zhongfeng capsule high-dose groups (1.08 g/kg), Zhongfeng capsule middle-dose groups (0.54 g/kg), Zhongfeng capsule low-dose groups (0.27 g/kg), with 10 rats in each group. Rats were treated with Zhongfeng capsule by gavage once a day for 10 days. The rats were sacrificed and the brain tissue was obtained after the experiment in each group. Score neurological deficit was evaluated after 24 h of the last intervention in rat of each group. The pathological changes of brain tissue were observed by HE staining. The serum levels of estradiol (E2) and follicle stimulating hormone (FSH) were determined by ELISA. The expressions of key genes and proteins of PI3K/Akt/Beclin1 signaling pathway in brain tissue were detected by qRT-PCR and Western blot respectively. Results: Compared with the sham operation group, the body weight and protein expressions of p-PI3k and p-Akt in brain tissue of rats were decreased significantly in the model group, while the brain index, neurological deficit score, gene and protein expressions of Beclin1 and LC3 were increased markedly in the model group(P＜0.05 or P＜0.01). In the model group, nerve cells of brain tissue were loosely packed, interstitial edema, triangular in shape, nuclear pyknosis and dark-blue staining were observed. Compared with the model group, the body weight of rats was increased obviously, the neurological deficit score was decreased significantly and the pathological injury of brain tissue was alleviated evidently in high-dose of Zhongfeng capsule group (P＜0.05). The brain index, the gene and protein expressions of Beclin1 and LC3 were decreased apparently in Zhongfeng capsule treatment groups(P＜0.05 or P＜0.01), while the expressions of p-PI3k and p-Akt in brain tissue were increased evidently in Zhongfeng capsule treatment groups(P＜0.05 or P＜0.01). Conclusion: Zhongfeng capsule can inhibit autophagy and improve brain neurons lesion of CIRI rats, the mechanism may be related to regulate the expression of Beclin1 and LC3 in PI3K/Akt/Beclin1 signaling pathway.
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Animals , Male , Rats , Autophagy-Related Proteins/pharmacology , Beclin-1/metabolism , Body Weight , Brain , Brain Ischemia/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Rats, Sprague-Dawley , Reperfusion Injury/drug therapyABSTRACT
ObjectiveTo develop a quantitative analysis of multi-components by single marker （QAMS） for determination of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills， and to provide a method for improving the national standard of the pills. MethodHigh performance liquid chromatography （HPLC） was developed for simultaneous determination of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills and the methodology validation was carried out. The chromatographic separation was performed on a Nucleosil 100-5 C18 column （4.6 mm×250 mm， 5 μm） with the mobile phase of acetonitrile -0.1% potassium dihydrogen phosphate aqueous solution （pH adjusted to 3.2 with phosphoric acid） （48∶52）， and the flow rate was 0.6 mL·min-1， the detection wavelength was set at 296 nm and the column temperature was 35 ℃. Taking cinobufagin as the internal standard， the relative correction factors （RCFs） of bufalin and resibufogenin were calculated， and the key influencing factors of RCFs were investigated. Relative retention time was used for the chromatographic peak location of the analyte， combining with the on-line ultraviolet spectroscopy and accurate relative molecular weight obtained by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-QTOF/MS）. The external standard method was used to verify the contents of three components obtained by QAMS. ResultQAMS was established for the determination of bufalin， cinobufagin and resibufogenin in the samples， and RCFs of cinobufagin to bufalin and resibufogenin were 0.922 and 1.01， respectively. The total content of the three marker compounds in 11 batches of Shexiang Baoxin pills was 33.7-36.0 µg per pill. There was no significant difference between the quantitative results of QAMS and external standard method. ConclusionThe established method can be used for the quality control of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills. It is suggested that bufalin should be considered as one of three marker compounds， and the sum of bufalin， cinobufagin and resibufogenin should be used for the content limit of this preparation.
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Objective:To determine the change in the number of family planning technical services in Pudong New Area of Shanghai from 2011 through 2018， and provide scientific evidence for improving reproductive health care for women of childbearing age. Methods:Trend analysis was performed according to the data of family planning technical service and obstetric service records by all medical and health care facilities in Pudong New Area from 2011 through 2018. Results:Top four family planning operations were vacuum aspiration， intrauterine device （IUD） removal， IUD insertion and medical abortion. The average annual number of family planning operations was 77 320， which declined over years with an average reduction rate of 3.32%. The average reduction rate of contraceptive and induced abortion was 5.87% and 1.6%， respectively. The reduction rate of IUD insertion was higher than that of IUD removal. Vacuum aspiration was the leading method of induced abortion， accounting for 77.9%， in which 79.27% was performed with intravenous anesthesia. Among the women receiving induced abortion， majority of them were married and non-local residents. The ratio of induced abortion to the live birth decreased over years from 2011 through 2018. Conclusion:Number of family planning technical services in Pudong New Area of Shanghai decreases from 2011 through 2018. Vacuum aspiration and IUD removal are the main operations in family planning surgery. With the decrease of long-term medically controlled contraceptive and birth control measures， it is necessary to strengthen the correct and effective use of self-controlled contraceptive measures for women of childbearing age， especially for those non-local married women， so as to reduce unwanted pregnancies and repeated induced abortions.
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As a new strategy capable of uncovering the characteristics of traditional Chinese medicines, the quantitative analysis of multi-components by single-marker(QAMS) has been widely employed for the quality evaluation of Chinese medicinal materials, slices, and extracts. However, its application in the assessment of Chinese patent medicines is yet to be explored. By referring to the determination of three bufogenins in Bufonis Venenum by QAMS described in Chinese Pharmacopoeia(2020 Edition), this paper selected seven representative preparations containing Bufonis Venenum and explored whether the relative correction factors(RCFs) of cinobufagin(CB) to bufalin(BF) and resibufogenin(RB) could be directly used for the quality control of Bufonis Venenum-contained preparations. Based on the qualitative analyses under the same chromatographic conditions as used for toad venom, combing specificity test, five preparations such as Yatong Yili Pills, Houzheng Pills, Xiongdan Jiuxin Pills, Liushen Pills and Niuhuang Xiaoyan Pills, were expected to use validated RCFs for the direct determination of three components. Taking Houzheng Pills as an example, the methodological validation of bufalin, cinobufagin and resibufogenin was carried out, and the recoveries of bufalin, cinobufagin and resibufogenin were 90.64%-106.1%. The obvious difference was not observed between the contents of bufalin and resibufogenin in 24 batches of preparation samples by QAMS and external reference method. In the tested samples, the content of bufalin, cinobufagin and resibufogenin were 1.27-2.61, 2.44-5.66 and 0.988-3.16 mg·g~(-1) in 10 batches of Liushen Pills samples. The contents of bufalin, cinobufagin and resibufogenin were 0.760-1.32, 1.35-2.39 and 0.600-1.55 mg·g~(-1) in 10 batches of Houzheng Pills samples from three manufacturers. The obtained data contribute to improving the quality standard of Bufonis Venenum-contained preparations, and they also provide some ideas for the application of QAMS in the quality evaluation and control of Chinese patent medicines.
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China , Chromatography, High Pressure Liquid , Medicine, Chinese Traditional , Nonprescription Drugs , Quality ControlABSTRACT
OBJECTIVE@#To investigate whether ginsenoside Rb1 (Rb1) can protect human umbilical vein endothelial cells (HUVECs) against high glucose-induced apoptosis and examine the underlying mechanism.@*METHODS@#HUVECs were divided into 5 groups: control group (5.5 mmol/L glucose), high glucose (HG, 40 mmol/L) treatment group, Rb1 (50 µ mol/L) treatment group, Rb1 plus HG treatment group, and Rb1 and 3-(@*RESULTS@#Rb1 ameliorated survival in cells in which apoptosis was induced by high glucose (P<0.05 or P<0.01). Upon the addition of Rb1, mitochondrial and intracellular reactive oxygen species generation and malondialdehyde levels were decreased (P<0.01), while the activities of antioxidant enzymes were increased (P<0.05 or P<0.01). Rb1 preserved the mitochondrial membrane potential and reduced the release of Cyt-c from the mitochondria into the cytosol (P<0.01). In addition, Rb1 upregulated mitochondrial biogenesis-associated proteins (P<0.01). Notably, the cytoprotective effects of Rb1 were correlated with SIRT3 signalling pathway activation (P<0.01). The effect of Rb1 against high glucose-induced mitochondria-related apoptosis was restrained by 3-TYP (P<0.05 or P<0.01).@*CONCLUSION@#Rb1 could protect HUVECs from high glucose-induced apoptosis by promoting mitochondrial function and suppressing oxidative stress through the SIRT3 signalling pathway.
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OBJECTIVE Cranial radiotherapy is clinically used in the treatment of brain tumors;however, the conse?quent cognitive and emotional dysfunctions seriously impair the life quality of patients. LW-AFC, an active fraction combi?nation extracted from classical traditional Chinese medicine prescription Liuwei Dihuang decoction, can improve cogni?tive and emotional dysfunctions in many animal models;however, the protective effect of LW-AFC on cranial irradiation-induced cognitive and emotional dysfunctions has not been reported. Recent studies indicate that impairment of adult hippocampal neurogenesis (AHN) and alterations of the neurogenic microenvironment in the hippocampus constitute crit?ical factors in cognitive and emotional dysfunctions following cranial irradiation. Here, our research further investigated the potential protective effects and mechanisms of LW-AFC on cranial irradiation-induced cognitive and emotional dys?functions in mice. METHODS LW-AFC (1.6 g·kg-1) was intragastrically administered to mice for 14 d before cranial irra?diation (7 Gyγ-ray). AHN was examined by quantifying the number of proliferative neural stem cells and immature neu?rons in the dorsal and ventral hippocampus. The contextual fear conditioning test, open field test, and tail suspension test were used to assess cognitive and emotional functions in mice. To detect the change of the neurogenic microenvi?ronment, colorimetry and multiplex bead analysis were performed to measure the level of oxidative stress, neurotrophic and growth factors, and inflammation in the hippocampus. RESULTS LW-AFC exerted beneficial effects on the contex?tual fear memory, anxiety behavior, and depression behavior in irradiated mice. Moreover, LW-AFC increased the num?ber of proliferative neural stem cells and immature neurons in the dorsal hippocampus, displaying a regional specificity of neurogenic response. For the neurogenic microenvironment, LW-AFC significantly increased the contents of superox?ide dismutase, glutathione peroxidase, glutathione, and catalase and decreased the content of malondialdehyde in the hippocampus of irradiated mice, accompanied by the increase in brain-derived neurotrophic factor, insulin-like growth factor-1, and interleukin-4 content. Together, LW-AFC improved cognitive and emotional dysfunctions, promoted AHN preferentially in the dorsal hippocampus, and ameliorated disturbance in the neurogenic microenvironment in irradiated mice. CONCLUSION LW-AFC ameliorates cranial irradiation-induced cognitive and emotional dysfunctions, and the underlying mechanisms are mediated by promoting AHN in the dorsal hippocampus and improving the neurogenic micro?environment. LW-AFC might be a promising therapeutic agent to treat cognitive and emotional dysfunctions in patients receiving cranial radiotherapy.
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@#Abstract】 【Objective】To investigate the effect and mechanism of ginsenoside Rb1 attenuating human umbilical vein endothelial cells(HUVEC) senescence induced by high glucose through Sirt3/SOD2 pathway.【Methods】The senescence of HUVEC induced by high glucose(40 mmol/L)was assessed by senescence-associated β-galactosidase(SA-β-Gal)staining,and the expression of plasminogen activator inhibitor 1(PAI-1)and P16. Annexin V-FITC/PI was performed to measure apoptotic effect. The expression of sirtuins 3(Sirt3)and superoxide dismutase 2(SOD2)was detected by western blot. Meanwhile,the level of intracellular malondialdehyde(MDA)and the activity of SOD2 were measured.【Results】Treatment of HUVEC with high glucose for 24 hours induced premature senescence instead of apoptosis,as indicated by a larger proportion of the cells stained with SA-β-Gal and the up-regulated expression of PAI-1 and P16. Pretreatment of HUVEC with ginsenoside Rb1(40 μmol/L)could reverse endothelial cell senescence,as indicated by the reduced SA-β-Gal positive cells and the down-regulated expression of PAI-1 and P16. Furthermore,ginsenoside Rb1 pretreatment upregulated the protein expression of Sirt3 and SOD2,and eventually increased the activity of SOD2 and decreased the level of MDA.【Conclusion】Ginsenoside Rb1 could antagonize high glucose-induced premature senescence of HUVEC via Sirt3/SOD2 signaling pathway.
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OBJECTIVE To explore the effect and mechanisms of LW-AFC,a new formula derived from Liu Wei Di Huang Tang,on irradiation-induced reduction of mice adult hippocampal neurogenesis. METHODS C57BL/6J mice were randomly divided into seven groups (n=10): control group, LW-AFC group (1.6 g·kg-1), Liu Wei Di Huang Tang (LW) group (10 g·kg-1), brain derived neurotrophic factor (BDNF) group, irradiation group, irradiation+LW group, and irradiation+LW-AFC group. Reduction of mice adult hippocampal neurogenesis was induced by cranial irradiation.LW-AFC was administered by oral gavage for 30 d after cranial irradiation treatment. Immunofluorescence and Nissl′s staining were performed for histological morphology assessment. Bromodeoxyuridine (BrdU) staining was used in the detection of proliferation cells. The peripheral blood and hippocampal homogenate were collected to measure the content of tumor necrosis factor-α (TNF-α), interferon-gamma (INF-γ), interleukin-1β (IL-1β),IL-4 and IL-10.The hippocampal homogenate was used for Western blot to detect the BDNF-TrkB signal pathway, including extracellular regulated protein kinases1/2 (ERK1/2) and BDNF target protein. Morris water maze and new object recognition test were performed to examine the cognitive function of mice.The mice forced swimming and tail suspension test were used to assess alteration in depressive behavior. Long term potentiation was used to examine the synaptic plasticity change of mice. RESULTS Adult hippocampal neurogenesis was significantly reduced after irradiation of 20 Gray dose (10 Gray per day, total 2 d). LW-AFC treatment increased the BrdU number of irradiated mice (P<0.05). In Morris water maze test, LW-AFC group showed decreased escape latency in the learning period (P<0.05), while increased the number of crossing the platform in the memory period. LW-AFC can also reduce the immobility time of mice in the tail suspension test (P<0.01). CONCLU-SION LW-AFC modulates adult neurogenesis to ameliorate cognitive impairment and reduce depres-sive behavior in radiation injury mice.
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Objective:To explore the diagnostic value of the Montreal cognitive assessment combined with delayed recall in time-limited screening for screening the elderly mild cognitive impairment (MCI) population.Method:A total of 178 elderly patients were enrolled in this study,and EADC was used to diagnose MCI.The patients were divided into normal group (n=92) and case group (n=86).178 cases were accepted by the conventional method of Montreal cognitive assessment and the Montreal cognitive assessment combined with delayed recall part of the time limit application,with the application of the receiver operating characteristic curve (ROC) for analysis,and getting the best cut-off point.Result:①The area under the ROC curve of the Montreal cognitive assessment was 0.851,the area of the Montreal cognitive assessment combined with the delayed recall in time-limited Screening was 0.917.The area under the ROC curve of the Montreal cognitive assessment combined with the delayed recall in time-limited screening was higher than that of the conventional method.The area under the ROC curve between the two ways was statistically significant,and the difference was statistically significant.②The best cut-off value of the Montreal cognitive assessment was 25,the sensitivity was 88.37％ and the specificity was 85.37％.The best cut-off value of the Montreal cognitive assessment combined with the delayed recall in time-limited screening was 25,the sensitivity was 95.35％ and the specificity was 78.26％.Conclusion:①The Montreal cognitive assessment Combined with delayed recall in time-limited screening can be applied to the diagnosis of mild cognitive impairment in the elderly.The diagnostic value of the Montreal cognitive assessment combined with the d elayed recall in time-limited screening was higher than that of the conventional method.②The cut-off point established by the ROC curve can be used as a diagnostic reference for the evaluation of mild cognitive impairment in the elderly as a part of the Montreal cognitive assessment combined with delayed recall in time-limited screening.
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[ ABSTRACT] AIM:To investigate the effects of sinapic acid ( SA) on the proliferation and apoptosis of rat vas-cular smooth muscle cells (VSMCs) induced by high glucose (HG).METHODS:Cultured A7r5 cells were randomly di-vided and treated as indicated.The cell viability was determined by MTT assay.DNA synthesis was measured by BrdU as-say.Cell cycle progression and cell apoptotic rate were determined by flow cytometry analysis.The levels of reactive oxygen species (ROS) were detected by ELISA.The protein levels of cyclin D1, P21, P27, phosphorylated protein kinase C (p-PKC), p-P38 andβ-actin were evaluated by Western blot.RESULTS:Compared with control group, the viability of A7r5 cells was significantly enhanced, the DNA synthesis was increased, the cell cycle progression was promoted, the levels of ROS were elevated, the cell apoptotic rate was reduced, the protein expression of P21 and P27 was decreased, and the pro-tein levels of cyclin D1, p-PKC and p-P38 were increased in HG group (all P<0.05).These effects were reversed by SA (0.1, 1 and 10 μmol/L) treatment in a dose-dependent manner (all P<0.05).Both P38 inhibitor SB203580 and PKC inhibitor chelerythrine significantly inhibit HG-induced PKC/P38 activation and cell viability ( P <0.05).CONCLU-SION:SA inhibits HG-induced VSMCs proliferation and promotes cell apoptosis via reducing PKC/P38 activation.
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The present study was designed to isolate and evaluate the antibacterial activity of the compounds from the whole plant of Euphorbia helioscopia L.. Various chromatographic techniques were used to isolate and purify the compound. The structure of the compound was elucidated on basis of spectral data ((1)H NMR, (13)C NMR, (1)H-(1)H COSY, HSQC, HMBC, NOESY, IR, and HR-ESI-MS). A new jatrophone-type diterpenoid (14α,15β-diacetoxy-3β-benzoyloxy-7β-nicotinoyloxy-9-oxo-jatropha-5E,11E-diene), named euphoheliosnoid E (1), was isolated from the whole plant of E. helioscopia L. Compound 1 showed significant anti-microbial activity against oral pathogens.
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Anti-Infective Agents , Pharmacology , Diterpenes , Chemistry , Pharmacology , Euphorbia , Chemistry , Molecular Structure , Mouth Diseases , Microbiology , Niacin , Chemistry , Pharmacology , Plant Extracts , Chemistry , PharmacologyABSTRACT
BACKGROUND:5-Fluorouracil occupies an important position in the treatment of gastric cancer, but its long-term use can easily induce adverse reactions such as myelosuppression and leukopenia. Polylactic acid and its copolymers have a higher biocompatibility, and their decomposer cannot gather in the body. OBJECTIVE:To investigate the in vitro cytotoxicity mechanism of 5-fluorouracil-loaded polylactic acid nanoparticles on gastric cancer cel lines. METHODS:Ten mice were selected in this study. 5-fluorouracil-loaded polylactic acid nanoparticles (1×10-7, 1×10-6, 1×10-5, 1×10-4 mol/L) were prepared using ultrasonic emulsification method. Kiling effect of polylactic acid nanoparticles on gastric cancer cel lines in vitrowere detected. Then, the inhibition rate was calculated at different concentrations. RESULTS AND CONCLUSIONS: Under the transmission electron microscope, 5-fluorouracil-loaded polylactic acid nanoparticles had good shape and relatively evenly distributed with no adhesions. After drug administration, the drug concentration was 50% at 24 hours and 62.9% at 72 hours. After 48 hours co-culture with single 5-fluorouracil or 5-fluorouracil-loaded polylactic acid nanoparticles, the viability of gastric cancer cels showed a decrease trend with the increase of drug concentrations, and moreover, 5-fluorouracil-loaded polylactic acid nanoparticles had a better cel inhibition ability than the single 5-fluorouracil (P < 0.05). The IC50value of 5-fluorouracil-loaded polylactic acid nanoparticles was significantly lower than that of 5-fluorouracil (P < 0.05). These findings indicate that polylactic acid nanoparticles as good drug carriers have a strong drug loading capacity and increase drug concentration in the body, but cannot reduce the biological activity of 5-fluorouracil, which provide new ideas for the treatment of gastric cancer.
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Objective To observe the clinical efficacy ofXing Nao Kai Qiao (brain-awakening and orifice-opening) needling plus thunder-fire moxibustion in treating vertebrobasilar ischemia.Method Fifty-five patients with vertebrobasilar ischemia were randomized into a treatment group of 26 cases and a control group of 29 cases. The treatment group was intervened byXing Nao Kai Qiao needling plus thunder-fire moxibustion, while the control group was by oral administration ofYangxue Qingnao granules. The blood flow of vertebrobasilar arteries were observed by using ultrasonic Doppler blood-flow detector before and after intervention, and the clinical efficacies were compared between the two groups.Result The recovery and markedly-effective rate and total effective rate were respectively 61.5% and 96.2% in the treatment group, versus 34.5% and 89.7% in the control group, and the differences were statistically significant (P<0.01,P<0.05). The VS and VD of vertebral arteries and VS of basilar arteries were significantly changed after intervention in the treatment group (P<0.05). The VS of basilar arteries was significantly changed after intervention in the control group (P<0.05). After treatment, The VS of vertebral and basilar arteries in the treatment group was significantly different from that in the control group (P<0.05).ConclusionXing Nao Kai Qiao needling plus thunder-fire moxibustion is an effective approach in treating vertebrobasilar ischemia.
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Objective To investigate the safety and efficacy of mechanical thrombectomy (MT) compared with In?tra-arterial Thrombolysis (IAT) treatment in patients with severe acute ischemic stroke (AIS) caused by large cerebral ar?tery occlusion. Method The patients with AIS caused by large cerebral artery occlusion and underwent MT or IAT from 2005 May to 2014 May was included. A retrospective analysis was conducted on the onset to emergency(OTE)time, emergency to acupuncture(ETA)time, acupuncture to recanalization (ATR) time, stroke severity as measured by the Na?tional Institutes of Health Stroke Scale (NIHSS) score, and site of arterial occlusion on magnetic resonance angiography (MRA). A comparison was made between MT and IAT patients in rates of recanalization, symptomatic intracranial bleed?ing (SIB), mortality, and functional outcome. Three-month favourable outcome was defined as a modified Rankin Scale (mRS) score≤2. Result One hundred and two AIS patients were treated with MT and 50 with IAT. There was no differ?ence between MT and IAT groups with regard to demographics, onset NIHSS score (13.37±6.95 vs. 12.70±6.11;P=0.572) and discharge NIHSS score (8.40 ± 6.69 vs. 7.53 ± 7.28, P= 0.522) and the change of NIHSS score (3.87 ± 7.14 vs. 4.26 ± 5.42, P=0.766). There were significantly differences between MT and IAT groups in the OTE time (Median 300 min vs. 120 min,Z=-5.704,P=0.000) , ATR time (Median 30 min vs. 65 min,Z=-5.011,P=0.001) ,recanalization (91.2%vs. 60.0%,P =0.01),the rate of AIB(21.7% vs. 36.0%,P =0.046),3-month mortality (16.6% vs. 26.0%,P =0.043). The above parameters were better in MT group than in the IAT group. There were no significant differences between MT and IAT groups in the rate of SIB (12% vs. 16%,P =0.055), the NIHSS change(Median 3 vs. 4,Z =-0.236,P =0.823) and mRS score on 90d ( 48.2%vs. 46.0%, P=0.823). MT patients had significantly higher percentages of stent use (22.5%vs. 8%,P=0.018) . The Recanalization for ICA(81.8%vs. 55.6%,P=0.048),BA(93.1%vs. 55.6%,P=0.032)and MCA( 97.5% vs. 60.0%,P =0.026)was higher in MT group than in IAT group .The SIB rate for ICA(13.8% vs. 33.3%,P =0.000),BA(13.8%vs. 33.3%,P=0.000)was lower in MT group than in IAT group . The mortality rate of was significant?ly lower in MT than in IAT group for MCA (2.5%vs. 20.0%,P=0.000) . the good outcome rate for BA was higher in MT group than in IAT group(41.3%vs. 22.2%,P﹤0.01). Conclusions Compared to IAT,MT can provide broader time win?dow,higher recanalization rate and better outcome in patients with severe acute ischemic stroke (AIS) caused by large ce?rebral artery occlusion.
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Objective To investigate the effect of Terpinen-4-ol on the proliferation of lung adenocarcinoma A-549 cells and its related mechanism. Methods A-549 cells were treated with different concentrations of Terpinen-4-ol. The inhibitory effect of Terpinen-4-ol on A-549 cells was tested by MTT method. Cell grow ability was determined by CCK-8 colorimetry. The ultrastructure of A549 cells were observed by transmission electron microscopy before and after Terpinen-4-ol treatment. The changes of cell cycle, apoptosis, and the level of intracel-lular calcium were inspected by flow cytometry. Inoculated the lung adenocarcinoma A-549 cells on the nude mice to form transplantation tumor. The experimental nude mice with transplantation tumors were divided into three groups:negative control group,high dose positive con-trol group and low dose positive control group. The mice were given continuously intraperitoneal injection for 10 days, and then the transplan-tation tumors were taken and the size and weight of them were detected. Results After Terpinen-4-ol treatment for 24 h,MTT assay showed that the IC50 value of A549 cells was 0. 067% v/v. The growth curves of positive control groups were significantly smooth than the negative control group. The formation of autophagosome increased after treatment with Terpinen-4-ol. The results of flow cytometry showed that the cell cycle was arrested in S phase,Terpinen-4-ol could induce apoptosis of A549 cell, The intracellular calcium concentrations in positive control groups were significantly higher than the negative control group(P<0. 05). Low dose group and high dose group restrained the growth of the transplantation tumor obviously, and the tumor inhibitory rate were 53. 33% and 77. 76% respectively. Conclusion Terpinen-4-ol has inhibitory effect on the proliferation of A-549 cells in vitro and in vivo.
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Objective Xerophthalmia is a common disease of the ocular surface .Lupus nephritis ( LN) may induce changes in the ocular surface , which has aroused increasing attention in ophthalmology .This study was to observe the morphological changes of the tear meniscus in LN patients with xerophthalmia after treated with artificial tears . Methods This clinical trial included 60 LN patients with xerophthalmia (120 eyes) and another 60 without.The xerophthalmia group received artificial tear eye-drops for 1 month. The tear meniscus height (TMH), tear meniscus depth (TMD), and tear meniscus area (TMA) were measured by anterior segment optical coherence tomography ( AS-OCT) before and after intervention . Results There were significant differences between the xe-rophthalmia and control groups in the baseline TMH ([0.159 ±0.051] vs [0.269 ±0.053] mm, P<0.01), TMD ([0.154 ± 0.012] vs [0.198 ±0.007] mm, P<0.01), and TMA ([0.021 ±0.069] vs [0.052 ±0.021] mm2, P<0.01).After 1 month of intervention with artificial tears, the xerophthalmia patients showed dramatically increased TMH ([0.179 ±0.014] vs [0.159 ± 0.051] mm, P<0.01), TMD ([0.163 ±0.015] vs [0.154 ±0.012] mm, P<0.01), and TMA ([0.040 ±0.017] vs [0.021 ± 0.069] mm2, P<0.01). Conclusion AS-OCT, with its advantages of clearly and objectively imaging and measuring the cross section of the tear meniscus , can be applied to the diagnosis and assessment of xerophthalmia in LN patients , and artificial tear eye-drops can effectively relieve their xerophthalmia symptoms .
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Obstructive sleep apnea (OSA) is characterized by repeated intermittent hypoxia (IH), hypercapnia, sleep fragmentation and intrathoracic pressure change. IH is related to the clinical pathophysiological processes of hypertension, atherosclerosis, coronary heart disease, arrhythmia, stroke, heart failure and sudden death. IH from OSA can lead to metabol-ic dysregulation, endothelial dysfunction, systemic inflammation, oxidative stress and the change of nerve body fluids, which has been shown to increase the risk of cardiovascular diseases. This study mainly describes the pathogenesis of IH leading to the various cardiovascular diseases.
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This study sought to investigate the in vivo antiviral effect of amantadine (AM) and biphenyl dimethyl dicarboxylate (DDB) on hepatitis B virus (HBV) in HBV replication mice. HBV replication-competent plasmid was transferred into male BALB/c mice by using hydrodynamics-based in vivo transfection procedure to develop HBV replication mouse model. The model mice were matched by body weigh, age and serum levels of hepatitis B e antigen (HBeAg) and were divided into four groups: AM group, DDB group, AM+DDB group and NS group, with the last one as control, and the mice of each group were administered corresponding agent orally twice a day, in a medication course lasting 3 d. On the third day, the mice were sacrificed 4-6 h after the last oral intake. HBV DNA replication intermediates in liver were analyzed by Southern blot hybridization. The serum hepatitis B surface antigen (HBsAg) and HBeAg were detected by enzyme linked immunosorbent assay (ELISA). Compared to the animals in the control group, HBV DNA replication intermediates in liver and HBsAg and HBeAg in serum from the AM and AM plus DDB group of mice decreased, and there was no difference between these two groups of mice. The levels of HBV DNA intermediate from liver and the serum HBsAg and HBeAg between the control and DDB group, however, were not obviously different. In conclusion, the inhibition effect of AM on HBV was detected, but treatment with DDB for 3 days did not influence the viral replication and expression of HBV in the HBV replication mice.
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Animals , Male , Mice , Amantadine , Pharmacology , Antiviral Agents , Pharmacology , DNA Replication , DNA, Viral , Dioxoles , Pharmacology , Disease Models, Animal , Hepatitis B , Virology , Hepatitis B Surface Antigens , Blood , Hepatitis B e Antigens , Blood , Hepatitis B virus , Physiology , Mice, Inbred BALB C , Plasmids , Transfection , Virus ReplicationABSTRACT
Background Phlyctenular ophthalmia is a delayed hypersensitivity reaction to some microprotein and affected mainly by adolescent in high incidence.Objective This study was to investigate the microscopy findings of phlyctenular ophthalmia and evaluate the histological changes by laser scanning confocal microscope.Methods Twenty-nine eyes suffered from phlyctenular ophthalmia and twenty normal eyes were examined using laser scanning confocal microscope.The pictures were taken by a CCD camera.All the cases had initially chest X-ray,tuberculin test,bacterial and mycobacteria culture.Results Dendritic and inflammatory cells were increased and concentrated in conjunctiva,and epithelial cells were deformed and squamatizated.The capillaries engorged and the goblet cells were injured.The corneoscleral Vogt meshing of the phlyctenular keratitis was obscured and dendritic cells were intruded into the corneas.The corneal epithelium of phlyctenular keratitis was absent and the subepithelial nerve plexus were bended and fractured,and the dendritic and inflammatory cells were intruded.Scarring of corneal stroma was seen under the laser scanning confocal microscope.Conclusions Laser scanning confocal microscopy is valuable for basic research and clinical diagnosis of phlyctenular ophthalmia.
ABSTRACT
<p><b>BACKGROUND</b>Molecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms (SNPs) of blood group antigens can be determined by the polymerase chain reaction with sequence specific priming (PCR-SSP) assay. Commercial high-throughput platforms can be expensive and are not approved in China. The genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE blood group antigens in Jiangsu province were unknown. The aim of this study is sought to detect the genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE antigens in Jiangsu Chinese Han using molecular methods with laboratory developed tests.</p><p><b>METHODS</b>DNA was extracted from EDTA-anticoagulated blood samples of 146 voluntary blood donors collected randomly within one month. Standard serologic assay for red blood cell antigens were also performed except the Scianna blood group antigens. PCR-SSP was designed to work under one PCR program to identify the following SNPs: JK1/JK2, KEL1/KEL2, FYA/FYB, SC1/SC2, C/c and E/e.</p><p><b>RESULTS</b>Serologic antigen results were identical to the phenotypes that were predicted from genotyping results. The allele frequencies for Jk*01 and Jk*02 were 0.51 and 0.49, respectively; for Fy*A and Fy*B 0.94 and 0.06; for RHCE*C and RHCE*c 0.68 and 0.32; and for RHCE*E and RHCE*e 0.28 and 0.72. Among 146 blood donors, all were KEL*02/KEL*02 and SC*01/SC*01, indicating allele frequencies for KEL*02 and SC*01 close to 1.00.</p><p><b>CONCLUSIONS</b>The use of PCR-SSP working under the same condition for testing multiple antigens at the same time is practical. This approach can be effective and cost-efficient for small-scale laboratories and in developing counties. These molecular tests can be also used for identifying rare blood types.</p>