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Objective:To observe the effects of Traditional Chinese Medicine (TCM)ultrasound drug permeation electrotherapy device on the inflammatory response of rats with cerebral ischemia, and to provide an experimental basis for the clinical application of TCM ultrasound drug permeation electrotherapy device in the treatment of cerebral ischemia.Methods:A total of 72 SD rats were randomly divided into sham-operation group (12 rats) and modeling group (60 rats). The middle cerebral artery occlusion (MCAO) model was prepared by thread embolism in the model group. The rats were divided into model group, Chinese medicine tablet group, blank tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "blank tablet + electrotherapy group"), Chinese medicine tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "Chinese medicine tablet + electrotherapy group") and butylphthalide group according to the random number table method, with 12 rats in each group. The corresponding treatment was given continuously for 7 days. The neurological function was scored using Longa method evaluation criteria; TTC staining was used to observe the infarct volume and calculate the percentage of infarct volume; HE staining was used to observe the cell morphology of cortical area in each group of rats; ELISA was used to detect the serum TNF-α and IL-1β levels in each group of rats; TLR4, MyD88 and NF-κBp65 protein expressions in hippocampal tissue of each group of rats on the infarct side were detected by Western blot method.Results:Compared with the model group, the neurological function scores of rats in the blank tablet + electrotherapy group, the herbal tablet + electrotherapy group, and the butylphthalein group significantly decreased ( P<0.05), the percentage of cerebral infarct volume significantly decreased ( P<0.05), the contents of serum TNF-α and IL-1β significantly decreased ( P<0.05), and the expressions of TLR4 (0.42±0.07, 0.31±0.07, 0.19±0.04 vs. 0.68±0.14), MyD88 (0.39±0.12, 0.30±0.07, 0.23±0.11 vs. 0.67±0.10), NF-κBp65 (0.32±0.03, 0.27±0.02, 0.17±0.03 vs. 0.57±0.12) protein in hippocampal tissue significantly decreased ( P<0.05). Conclusion:The TCM ultrasound drug permeation electrotherapy device can inhibit TLR4, MyD88, NF-κBp65 protein expressions and reduce the release of serum inflammatory factors TNF-α and IL-1β, thus exerting cerebral ischemic protective effects.
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CRISPR, as an emerging gene editing technology, has been widely used in multiple fields due to its convenient operation, less cost, high efficiency and precision. This robust and effective device has revolutionized the development of biomedical research at an unexpected speed in recent years. The development of intelligent and precise CRISPR delivery strategies in a controllable and safe manner is the prerequisite for translational clinical medicine in gene therapy field. In this review, the therapeutic application of CRISPR delivery and the translational potential of gene editing was firstly discussed. Critical obstacles for the delivery of CRISPR system in vivo and shortcomings of CRISPR system itself were also analyzed. Given that intelligent nanoparticles have demonstrated great potential on the delivery of CRISPR system, here we mainly focused on stimuli-responsive nanocarriers. We also summarized various strategies for CIRSPR-Cas9 system delivered by intelligent nanocarriers which would respond to different endogenous and exogenous signal stimulus. Moreover, new genome editors mediated by nanotherapeutic vectors for gene therapy were also discussed. Finally, we discussed future prospects of genome editing for existing nanocarriers in clinical settings.
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New drug discovery is under growing pressure to satisfy the demand from a wide range of domains, especially from the pharmaceutical industry and healthcare services. Assessment of drug efficacy and safety prior to human clinical trials is a crucial part of drug development, which deserves greater emphasis to reduce the cost and time in drug discovery. Recent advances in microfabrication and tissue engineering have given rise to organ-on-a-chip, an in vitro model capable of recapitulating human organ functions in vivo and providing insight into disease pathophysiology, which offers a potential alternative to animal models for more efficient pre-clinical screening of drug candidates. In this review, we first give a snapshot of general considerations for organ-on-a-chip device design. Then, we comprehensively review the recent advances in organ-on-a-chip for drug screening. Finally, we summarize some key challenges of the progress in this field and discuss future prospects of organ-on-a-chip development. Overall, this review highlights the new avenue that organ-on-a-chip opens for drug development, therapeutic innovation, and precision medicine.
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ObjectiveTo investigate the mechanism of Xumingtang in Gu Jin Lu Yan （《古今录验》） in regulating cell pyroptosis through the hypoxia-inducible factor-1α （HIF-1α）/NOD-like receptor pyrin domain-containing protein 3 （NLRP3） pathway in ischemic stroke （IS）. MethodSD rats were randomly divided into a sham operation group， a model group， low- and high-dose Xumingtang groups， and a metformin group， with 20 rats in each group. Oral administration was performed for 3 days， and tissue samples were collected. Differential messenger RNA （mRNA） was screened using high-throughput sequencing， and Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analyses were performed on key differentially expressed genes. The modified neurological severity score （mNSS） and 2，3，5-triphenyltetrazolium chloride （TTC） staining were used to evaluate the effect of brain infarction. Hematoxylin-eosin （HE） staining was used for pathological morphological observation of brain tissue. Enzyme-linked immunosorbent assay （ELISA） was used to compare the levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in the ischemic cortical region. Double staining immunohistochemistry was used to detect the co-localization of HIF-1α and NLRP3. Real-time quantitative polymerase chain reaction （PCR） was performed to detect the mRNA expression of NLRP3， HIF-1α， Caspase-1 （CASP-1）， and gasdermin D （GSDMD）. Western blot was used to detect the protein expression of HIF-1α， NLRP3， CASP-1， and GSDMD. ResultA total of 5 705 differentially expressed genes （2 733 downregulated and 2 972 upregulated） were obtained by mRNA sequencing. After conversion to homologous genes and intersection with the pyroptosis gene set， 95 key differentially expressed pyroptosis genes were obtained. Compared with the sham operation group， the model group showed significantly increased mNSS scores， larger brain infarction areas （P<0.01）， diverse neuronal morphology， disordered arrangement， widened cell gaps， significantly increased levels of IL-1β and IL-18 in the ischemic cortical region （P<0.01）， enhanced co-localization fluorescence intensity， and significantly increased mRNA and protein expression levels of HIF-1α， NLRP3， CASP-1， and GSDMD （P<0.01）. Compared with the model group， the high-dose Xumingtang group showed the most significant improvement in neurological function scores and brain infarction areas （P<0.01）. The neuronal integrity and arrangement were more complete， and the cell gaps were narrower in all groups with drug treatment， with significantly reduced co-localization fluorescence intensity. Xumingtang could reduce the levels of IL-1β， IL-18， and the mRNA and protein expression of HIF-1α， NLRP3， CASP-1， and GSDMD （P<0.05， P<0.01）， with the high-dose Xumingtang group showing the most significant effect （P<0.01）. ConclusionXumingtang in Gu Jin Lu Yan can inhibit cell pyroptosis and promote neurological function recovery after IS， which may be related to the inhibition of the HIF-1α/NLRP3 pathway.
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Objective:To investigate the effects of peripartum administration of low-dose corticosteroids or intravenous immunoglobulin (IVIG) on delivery outcomes in pregnant patients with primary immune thrombocytopenia (ITP).Methods:This prospective cohort study involved pregnant women (≥34 gestational weeks) who were diagnosed with ITP in Peking University People's Hospital from January 2017 to December 2021. Their platelet counts were between 20×10 9/L to 50×10 9/L without bleeding and none of them had been treated with any medications. All patients were divided into medication group (prednisone or IVIG) and platelet transfusion group based on their preference. Differences in vaginal delivery rate, postpartum hemorrhage rate and platelet transfusion volume between the two groups were compared using t-test, Wilcoxon rank sum test and Chi-square test. Binary logistic regression was used to investigate the factors influencing the rates of vaginal delivery and postpartum hemorrhage. Multiple linear regression was used to analyze the factors influencing the platelet transfusion volume. Results:A total of 96 patients with ITP were recruited with 70 in the medication group and 26 in the platelet transfusion group. The vaginal delivery rate in the medication group was higher than that in the platelet transfusion group [60.0% (42/70) vs 30.8% (8/26), χ 2=6.49, P=0.013]. After adjusted by the proportion of multiparae and the gestational age at delivery, binary logistic regression showed that the increased vaginal delivery rate in patients undergoing the peripartum treatment ( OR=4.937, 95% CI: 1.511-16.136, P=0.008). The incidence of postpartum hemorrhage in the two groups was 22.9% (16/70) and 26.9% (7/26), respectively, but no significant difference was shown ( χ 2=0.17, P=0.789). The median platelet transfusion volume was lower in the medication group than in the platelet transfusion group [1 U(0-4 U) vs 1 U(1-3 U), Z=－2.18, P=0.029]. After adjustment of related factors including the platelet count at enrollment, obstetrical complications and anemia, multiple linear regression showed that the platelet transfusion volume was also lower in the medication group (95% CI:0.053-0.911, P=0.028). Ninety-six newborns were delivered without intracranial hemorrhage. The overall incidence of neonatal thrombocytopenia was 26.0% (25/96). There was no significant difference in birth weight, and incidence of neonatal asphyxia or thrombocytopenia between the two groups. Conclusion:Peripartum therapy in ITP patients may increase vaginal delivery rate and reduce platelet transfusion volume without causing more postpartum hemorrhage.
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Objective:To prepare vorinostat encapsulated hydroxypropyl-β-cyclodextrin (SAHA-CD) eye drops and investigate its inhibitory effect on corneal neovascularization (CNV) induced by alkali burns in mouse.Methods:The SAHA-CD eye drops at concentrations of 0.1%, 0.2%and 0.4%were prepared by inclusion technology with hydroxypropyl-β-cyclodextrin, and the content was assayed by high performance liquid chromatography.Seventy-five SPF mice with alkali burn-induced CNV were randomized into 0.1%SAHA-CD group, 0.2%SAHA-CD group, 0.4%SAHA-CD group, dexamethasone group and normal control group according to a random number table, 15 for each group, among which the SAHA-CD groups and dexamethasone group were treated with corresponding drugs, and model control group was treated with normal saline immediately after modeling, four times a day and five microliters each time, lasting for six days.The healing of corneal epithelium was examined with a slit lamp microscope after fluorescein sodium staining, and the areas of cornea epithelial defects were calculated using Eyestudio software.The corneal flat mount was prepared, and the length and areas of CNV were calculated with ImageJ software.The histology of mouse corneas was observed through hematoxylin and eosin staining.The expression level of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and matrix metalloproteinase-9 (MMP-9) in cornea were measured with enzyme linked immunosorbent assay (ELISA) kits.The use and care of animals complied with the ARVO statement and this study protocol was approved by the Experimental Animal Ethics Committee of Henan Eye Institute (No.HNEECA-2020-01).Results:The actual drug contents of the 0.1%, 0.2% and 0.4%SAHA-CD eye drops were 97.62%, 98.33%and 98.14%of the labeled amount.The cornea showed edema and opacification after modeling.On the sixth day after treatment, significant differences were found in the length and areas of CNV among various groups ( F=7.655, 8.802; both at P<0.01).The areas of CNV in 0.2%SAHA-CD, 0.4%SAHA-CD and dexamethasone groups were significantly smaller than model control group, and the length of CNV in 0.1%SAHA-CD, 0.2%SAHA-CD and dexamethasone groups were significantly smaller than model control group (all at P<0.05).On the third and sixth day following modeling, significant differences in the expression levels of VEGF, bFGF and MMP-9 were found among the five groups (third day: F=6.345, 7.149, 18.650; all at P<0.01; sixth day: F=6.749, 5.105, 5.023; all at P<0.01), and the expression levels of VEGF, bFGF and MMP-9 in 0.2%SAHA-CD group were significantly lower than those in 0.1%SAHA-CD group, 0.4%SAHA-CD group and model control group (all at P<0.05). Conclusions:SAHA-CD eye drops can inhibit alkali burn-induced CNV in mouse.
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Objective@#To understand the online learning related screen use duration and screen types in school aged children in Shanghai during the COVID-19 epidemic.@*Methods@#Random clustering sampling was used to select 5 591 parents of students from 8 primary and junior schools that are in the sampling pool of the national myopia survey in districts of Jiading, Pudong and Baoshan in Shanghai in April 2020. Electronic questionnaire was administered to parents regarding their child s online learning related screen use.@*Results@#On average, the median weekly duration of online learning related screen use was 13.33 hours, the curricular and extracurricular parts of which were 10(8.75，16.67) and 0(0，3.33) hours, respectively. About 29.44% of investigated school aged children only used small size screen for online learning. Children in higher grades, being myopic and parents neither being myopic were associated with reporting higher weekly duration( P <0.05); children in higher grades of primary school and parents neither being myopic were associated with a higher likelihood of using small size screen for online learning( P <0.05).@*Conclusion@#At the early stage of the COVID-19 epidemic, the burden associated with online learning related screen use was high in school aged children in Shanghai. Health education regarding online learning related screen use should be addressed in parents to guide their children to use screen appropriately.
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Objective@#To analyze the effect of sunlike spectrum LED illumination on retinal blood flow perfusion, and to explore the the correlation between sunlike spectrum LED illumination and eye health indicators in children and adolescents.@*Methods@#A randomized control double blind trial was conducted. The ordinary LED table lamp in the control group(11) and the sunlike spectrum LED table lamp in the experimental group(12) had a fitting degree of 87% and 95% with the daylighting spectrum, respectively. Two sample independent t test and multivariable linear regression model were applied to compare the changes of retinal blood perfusion before and after the trial.@*Results@#After near reading for 1 hour, the retinal capillary density in the superficial and deep layers of the subjects in the ordinary LED illumination group decreased (superficial layer: -3.05±2.04 , P <0.01; deep layer: -4.03± 4.94, P =0.02), no significant decrease was found in the sunlike spectrum LED illumination group (superficial layer: -0.59± 1.44, P =0.18; deep layer: -0.49±4.27, P =0.70). Multivariable regression analysis found that compared with ordinary LED illumination, sunlike spectrum LED illumination could significantly alleviate the decrease in capillary density in the superficial and deep retinal layers, respectively ( β =2.83, 95% CI =1.54-4.12, P <0.01; β =4.21,95% CI =0.58-7.84, P =0.02).@*Conclusion@#Sunlike spectrum LED illumination can alleviate the decrease in retinal blood perfusion caused by near work among children and adolescents, suggesting that it may delay the onset and development of myopia. Prevention and control of myopia needs to pay attention to the spectral power distribution of artificial illumination.
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Astrocytes are increasingly recognized to play an active role in learning and memory, but whether neural inputs can trigger event-specific astrocytic Ca2+ dynamics in real time to participate in working memory remains unclear due to the difficulties in directly monitoring astrocytic Ca2+ dynamics in animals performing tasks. Here, using fiber photometry, we showed that population astrocytic Ca2+ dynamics in the hippocampus were gated by sensory inputs (centered at the turning point of the T-maze) and modified by the reward delivery during the encoding and retrieval phases. Notably, there was a strong inter-locked and antagonistic relationship between the astrocytic and neuronal Ca2+ dynamics with a 3-s phase difference. Furthermore, there was a robust synchronization of astrocytic Ca2+ at the population level among the hippocampus, medial prefrontal cortex, and striatum. The inter-locked, bidirectional communication between astrocytes and neurons at the population level may contribute to the modulation of information processing in working memory.
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Animals , Humans , Mice , Astrocytes , Hippocampus/physiology , Memory, Short-Term/physiology , Neurons/physiology , Population DynamicsABSTRACT
Objective:To understand the progress, maternal morbidity, and maternal and infant outcomes in pregnant women with non-severe primary immune thrombocytopenia (ITP) during two consecutive pregnancies.Methods:This study retrospectively analyzed the clinical data of 40 patients with non-severe ITP who had two pregnancies and were treated at Peking University People's Hospital between June 2010 and June 2020. Platelet counts at different stages of pregnancy, treatments, maternal complications and neonatal outcomes were compared with Chi-square test, Fisher's exact test, paired sample t-test, non-parametric Wilcoxon signed rank test, independent sample t-test or non-parametric Mann-Whitney U test. Results:Among the 40 patients, 18 were diagnosed before and 22 were first diagnosed during the first gestation. Platelet counts and treatments in the 18 patients prior to their first conception were not significantly different from those in the 40 patients before their second pregnancy (all P>0.05). No significant difference in the average platelet count and thrombocytopenia severity at each stage of pregnancy, and maternal bleeding score or drug treatment was observed between the two pregnancies (all P>0.05), neither in the incidence of gestational hypertension, gestational diabetes, premature rupture of membranes, premature delivery, or anemia (all P>0.05). The incidences of postpartum hemorrhage and severe postpartum hemorrhage in the second pregnancy were 30.0%(12/40) and 22.5%(9/40), respectively, which were both higher than those in the first gestation [(7.5%(3/40) and 5.0%(2/40); χ2=6.64, 5.17; P=0.010, 0.023]. The amount of postpartum hemorrhage was higher in the second pregnancy than in the first [500 ml(213-795 ml) vs 300 ml(163-400 ml), Z=－2.34, P=0.019]. There was no significant difference in birth weight, the incidence of passive ITP or intracranial hemorrhage, or mortality between the neonates of the first and second pregnancy group (all P>0.05). The lowest platelet count in neonates within one week after birth in the second pregnancy group was (202.2±106.7)×10 9/L, which was lower than that of the first [(222.5±91.8)×10 9/L, Z=－2.04, P=0.041]. Conclusions:Non-severe ITP is not worse in the second pregnancy than in the first. In women with non-severe ITP, the incidence of maternal complications is not increased in the second pregnancy, but the risk of postpartum hemorrhage and the incidence of neonatal passive immune thrombocytopenia are raised.
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Objective:To explore the progression of paroxysmal nocturnal hemoglobinuria (PNH) during pregnancy and treatment for improving maternal and infant outcomes.Methods:Nine pregnant women with PNH were admitted to the Obstetrics Department of Peking University People's Hospital from September 2010 to September 2020. The clinical data of these patients were retrospectively collected and analyzed. Relevant literature was reviewed to summarize the progression, treatment, complications, perinatal outcomes, and follow-up of PNH during pregnancy. Descriptive methods were used for statistical analysis.Results:Among the nine patients, six were classic PNH, and three combined with bone marrow failure disease. Eight cases received blood transfusion/low-dose corticosteroids or combination therapy during pregnancy, and four of them were also received anticoagulants. In seven out of the eight patients diagnosed prenatally, the disease worsened during pregnancy. Complications were noted in eight patients during pregnancy, including fetal growth restriction in seven cases, hypertension and premature delivery in four cases each, thrombosis and intrauterine fetal death in one case each. No maternal deaths were reported, with a live birth rate of 8/9 between 33-38 gestational weeks, with the median at 37 weeks. The median time of postpartum follow-up was 50 months (4-92 months), during which the patients' conditions were all stable, and no abnormalities were found in the growth and development of the babies.Conclusions:Pregnant women with PNH tend to worsen with an increased incidence of perinatal complications and adverse outcomes. Multidisciplinary management is recommended for this population may help improve maternal and infant outcomes.
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Objective:To investigate the correlation of interferon (IFN) -γ rs2430561 single nucleotide polymorphisms (SNPs) and serum IFN-γ levels with susceptibility to herpes zoster.Methods:Blood samples were collected from 74 patients with herpes zoster and 40 healthy controls in General Hospital of Southern Theatre Command and the Fifth People Hospital of Hainan Province from November 2019 to November 2020. PCR and Sagner resequencing were conducted to detect the IFN-γ rs2430561 SNPs in the subjects, fluorescence-based quantitative PCR was performed to determine the copy number of varicella-herpes zoster virus (VZV) DNA in the serum of the patients with herpes zoster, and enzyme-linked immunosorbent assay was conducted to detect the serum IFN-γ level. Measurement data were compared by using t test or non-parametric test, and enumeration data by using chi-square test or Fisher′s exact test. Results:As rs2430561 genotyping showed, there were 4 patients with AA genotype, 37 with TT genotype and 33 with TA genotype in the herpes zoster group, as well as 13 subjects with TA genotype and 27 with TT genotype in the healthy control group, and the frequency of A allele of rs2430561 was significantly higher in the herpes zoster group (27.70%) than in the control group (16.25%, P=0.036) . The serum IFN-γ level ( M[ P25, P75]) was significantly lower in the herpes zoster group (33.45[0.80, 95.01]pg/ml) than in the control group (67.83[2.74, 318.35]pg/ml, U=1 822, P=0.028) . The VZV DNA copy number (expressed as a logarithm to the base of 10) per milliliter was 3.23 ± 0.71 in the serum of the patients with herpes zoster, and the serum IFN-γ level was negatively correlated with the VZV DNA copy number ( r=-0.302, P=0.009) . Conclusion:The carriage of rs2430561 A allele may affect the expression of IFN-γ, leading to higher susceptibility to herpes zoster.
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Cardiac hypertrophy is a common physiological or pathological process, and pathological cardiac hypertrophy can lead to heart failure, sudden death, etc. The role of microRNA (miRNA or MIR) in myocardial hypertrophy has gradually attracted public attention. miR-1 plays a certain protective role in the occurrence of cardiac hypertrophy. miR-133 is a key factor in the establishment of mast gene program, which is very important for the development of myocardial hypertrophy. Carvedilol and other drugs can regulate the expression of miR-133. miR-208a plays an important physiological role in the cardiovascular system, and its expression level changes dynamically in a variety of cardiovascular diseases such as cardiac hypertrophy, which is closely related to the progression and prognosis of the disease. The expression of miR-199a is up-regulated in pressure-overload cardiac hypertrophy, and it is found that miR-199a can inhibit autophagy of cardiomyocytes and induce the occurrence of cardiac hypertrophy. miR-200c can protect cardiomyocytes through a variety of pathways. miRNA may become an important biomarker or drug therapeutic target for cardiac hypertrophy. With the deepening of the research on non-coding RNAs including miRNA, its regulation on the occurrence of cardiac hypertrophy and the pathological process of heart failure will be further revealed.
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Objective:To investigate the pharmacokinetics of econazole solid lipid nanoparticles (E-SLNs) after administration of one single dose in rabbit eyes.Methods:E-SLNs with 0.2% econazole was prepared by microemulsion method.Its antifungal activity against Fusarium isolated from the eyes of patients with fungal keratitis was evaluated in vitro and was compared with natamycin eye drops.Four healthy New Zealand white rabbits were assigned to the blank control group without any drug interference during the experimental period, and other matched 21 rabbits were randomized into 7 groups according to the specimen-collected time, with 3 rabbits in each group.E-SLNs of 50 μl was singly applied to conjunctival sac in both eyes in the 21 rabbits, and tear was collected using a filter paper at 5, 15, 30, 60, 90, 120 and 180 minutes following administration of the drug.The cornea specimen was collected at above-mentioned time points respectively.The drug levels in each sample were assayed by high performance liquid chromatography.The accuracy, recovery rate, stability and antifungal activity of the drugs in tear fluid and cornea were detected.This study protocol was approved by the Life Science Ethics Review Committee of Henan Eye Hospital (No.HENNCA-2017-22). Results:For the tear samples and corneal tissue samples, the relative standard deviation ( RSD) of the accuracy of the drug was 2.34%-4.04%; the stability analysis result showed that the RSD of the drugs was less than 10%.The 50% minimum inhibitory concentration (MIC 50) and 90% minimum inhibit concentration (MIC 90) of E-SLNs were 0.37 μg/ml and 0.89 μg/ml, respectively.The MIC 50 and MIC 90 of natamycin were 1.15 μg/ml and 1.70 μg/ml, respectively.After one single dose application of E-SLNs eye drops, the peak time of the drug in tears fluids and cornea of rabbits were 5 minutes and maximum concentrations in tears and cornea were 597.64 μg/g and 33.15 μg/g, respectively. Conclusions:The drug levels in tears and cornea achieved are higher than MIC against Fusarium.
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Objective:To study the pharmacokinetics of the broad-spectrum antifungal drug butenafine nanomicelles (BTF-NM) after topical instillation.Methods:The self-assembly method was used to prepare BTF-NM.The particle size, Zeta potential, and polydispersity index (PDI) of BTF-NM were measured by a nano-particle size-Zeta potential analyzer, and the encapsulation efficiency was determined by high-performance liquid chromatography (HPLC). Forty-two healthy New Zealand white rabbits without eye disease were randomly divided into the BTF-NM group and the BTF suspension (BTF-S) group.The corresponding drugs were instilled in the conjunctival sac with a single instillation of 50 μl.The 7.5 mm filter paper was placed in the conjunctival sac of rabbit eye for 1 minute at 5, 15, 30, 60, 120, 180, 240 minutes after the administration.Then the rabbits were sacrificed by intravenous injection of 4% sodium pentobarbital solution through the ears of the rabbits.The aqueous humor was extracted and the corneal tissue was dissected.The drug concentration of BTF in different tissues was measured by HPLC.The study was approved by the Life Science Ethics Review Committee of Henan Eye Hospital (No.HNEECA-2019-01).Results:The particle size and PDI of BTF-NM were (15.65±0.04)nm and 0.11±0.01, respectively, the Zeta potential was (-0.29±0.36)mV, the encapsulation rate was (98.38±0.29)%.The peak time of the drug both in tears and corneal tissues after BTF-NM application was 5 minutes.The peak concentrations of the drug in tears and corneas of the BTF-NM group were (485.21±66.29) μg/g and (12.53±2.32) μg/g, which were 5.6 and 78 times than that of the BTF-S group, respectively.Within the observation time, the mass fractions of the drug in tears and corneas of the BTF-NM group at each time point were significantly higher than those of BTF-S group at corresponding time points (all at P<0.01), respectively.The area under the concentration-time curve (AUC) 0-240 minutes in tears and corneas of the BTF-NM group was 7 488.90 (μg/g)·minute and 829.01 (μg/g)·minute, which was 7.2 and 52 times than that of the BTF-S group, respectively.No drugs were detected in the aqueous humor of the BTF-NM group and the BTF-S group. Conclusions:BTF-NM is an ideal agent with a simple preparing process, high drug encapsulation efficiency and small particle size.Compared with BTF suspension, BTF-NM can significantly improve the bioavailability of BTF in rabbit corneas.
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Objective@#To observe changes in expression of autophagy proteins in peripheral CD4+ T lymphocytes and the epidermis of skin lesions, as well as generation of autophagy vesicles in epidermal cells in skin lesions of patients with herpes zoster, and to explore the relationship between varicella-herpes zoster virus (VZV) infection and autophagy.@*Methods@#Totally, 35 patients with herpes zoster were enrolled from Department of Dermatology, General Hospital of Southern Theater Command of PLA between December 2017 and December 2018, including 20 males and 15 females. Their age ranged from 18 to 79 (59.23 ± 9.27) years, pain duration was 5.14 ± 2.28 days, and lesion duration (from the onset of the lesion to the clinic visit) was 3.45 ± 1.77 days. Flow cytometry was performed to determine the expression of autophagy proteins including microtubule-associated protein 1 light chain 3B (LC3B) , Beclin-1 and p62 in peripheral blood CD4+ T lymphocytes of these patients. Thirty healthy adults served as control group. Lesional skin tissues were obtained from 12 patients with herpes zoster, and perilesional normal skin tissues of the same patient served as the control. Immunohistochemical study was conducted to determine the expression of autophagy proteins LC3B, Beclin-1 and p62 in epidermal tissues, and transmission electron microscopy to observe the generation of autophagy vesicles in epidermal cells. Two independent-sample t-test was carried out for intergroup comparison.@*Results@#The expression rates of autophagy proteins LC3B and Beclin-1 in peripheral CD4+ T lymphocytes were significantly higher in the herpes zoster group (61.23% ± 7.61%, 35.84% ± 4.22%, respectively) than in the control group (36.56% ± 4.27%, 15.34% ± 1.89%, respectively; t = 15.75, 24.56 respectively, both P < 0.01) , while the expression rate of p62 (5.75% ± 0.67%) was significantly lower in the herpes zoster group than in the control group (10.03% ± 1.15%, t = 18.65, P < 0.01) . Among the 12 patients with herpes zoster, the expression levels of LC3B and Beclin-1 in the epidermis were significantly higher in the skin lesions than in the perilesional normal skin tissues (t = 2.86, 4.58, P < 0.05) , but the expression level of p62 was significantly lower in the skin lesions than in the perilesional normal skin tissues (t = 2.43, P < 0.05) . Transmission electron microscopy showed formation of autophagy vesicles containing virus particles in epidermal cells in the skin lesions of 12 patients with herpes zoster, and vesicle counts were significantly higher in the skin lesions than in perilesional normal skin tissues (t = 9.67, P < 0.01) .@*Conclusion@#The autophagy level was elevated in peripheral CD4+ T lymphocytes and epidermis of skin lesions of patients with herpes zoster.
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Objective To observe changes in expression of autophagy proteins in peripheral CD4+ T lymphocytes and the epidermis of skin lesions,as well as generation of autophagy vesicles in epidermal cells in skin lesions of patients with herpes zoster,and to explore the relationship between varicella-herpes zoster virus (VZV) infection and autophagy.Methods Totally,35 patients with herpes zoster were enrolled from Department of Dermatology,General Hospital of Southern Theater Command of PLA between December 2017 and December 2018,including 20 males and 15 females.Their age ranged from 18 to 79 (59.23 ± 9.27) years,pain duration was 5.14 ± 2.28 days,and lesion duration (from the onset of the lesion to the clinic visit) was 3.45 ± 1.77 days.Flow cytometry was performed to determine the expression of autophagy proteins including microtubule-associated protein 1 light chain 3B (LC3B),Beclin-1 and p62 in peripheral blood CD4 + T lymphocytes of these patients.Thirty healthy adults served as control group.Lesional skin tissues were obtained from 12 patients with herpes zoster,and perilesional normal skin tissues of the same patient served as the control.Immunohistochemical study was conducted to determine the expression of autophagy proteins LC3B,Beclin-1 and p62 in epidermal tissues,and transmission electron microscopy to observe the generation of autophagy vesicles in epidermal cells.Two independent-sample t-test was carried out for intergroup comparison.Results The expression rates of autophagy proteins LC3B and Beclin-1 in peripheral CD4 + T lymphocytes were significantly higher in the herpes zoster group (61.23％ ± 7.61％,35.84％ ± 4.22％,respectively) than in the control group (36.56％ ± 4.27％,15.34％ ± 1.89％,respectively;t =15.75,24.56 respectively,both P ＜ 0.01),while the expression rate of p62 (5.75％ ± 0.67％) was significantly lower in the herpes zoster group than in the control group (10.03％ ± 1.15％,t =18.65,P ＜ 0.01).Among the 12 patients with herpes zoster,the expression levels of LC3B and Beclin-1 in the epidermis were significantly higher in the skin lesions than in the perilesional normal skin tissues (t =2.86,4.58,P ＜ 0.05),but the expression level of p62 was significantly lower in the skin lesions than in the perilesional normal skin tissues (t =2.43,P ＜ 0.05).Transmission electron microscopy showed formation of autophagy vesicles containing virus particles in epidermal cells in the skin lesions of 12 patients with herpes zoster,and vesicle counts were significantly higher in the skin lesions than in perilesional normal skin tissues (t =9.67,P ＜ 0.01).Conclusion The autophagy level was elevated in peripheral CD4+ T lymphocytes and epidermis of skin lesions of patients with herpes zoster.
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Yeast autolysis under solid-state fermentation can effectively promote the release of various active substances, thereby improving the quality of yeast products. The optimal process for yeast autolysis under solid-state fermentation was obtained by optimizing the autolysis temperature, autolysis time and the zinc ion concentration. We analyzed the indexes of free amino acid, soluble protein and α-amino nitrogen in the fermentation material, as well as A₂₆₀/A₂₈₀ ratio to determine yeast autolysis process conditions in the solid-state fermentation. On the basis of the obtained data, L₉ (3³) orthogonal test was designed to optimize the solid-state fermentation parameters for yeast autolysis: temperature at 40, 50 and 55 °C; time 12, 18 and 24 h; zinc ion concentration 2, 4 and 8 mg/kg. The optimum process conditions for yeast autolysis were: autolysis temperature 55 °C, time 18 h, zinc ion concentration 2 mg/kg, and soluble protein content reached 9.31 mg/g, free amino acid 14.36 mg/g, α-amino nitrogen 10.16 μg/g and A₂₆₀/A₂₈₀ 1.73. After optimization of the process, the soluble protein, free amino acid and α-amino nitrogen contents of the yeast autolysis production can be significantly increased, thereby obviously improving the quality of the composite culture.
Subject(s)
Humans , Amino Acids , Autolysis , Fermentation , Hydrogen-Ion Concentration , Nitrogen , Saccharomyces cerevisiae , TemperatureABSTRACT
Overexpression of CCND1 and t (11;14) (q13;q32) chromosomal translocation are important markers in mantle cell lymphoma (MCL). However, part of MCL lacks the expression of CCND1. SOX11 can be used as a biomarker for its overexpression in CCND1-negative MCL. SOX11 is a neurogenic transcription factor, and its overexpression is closely related to histone modification and DNA methylation. Differential expression of SOX11 in MCL is closely related with the plasma cell differentiation, and there is a relationship between the prognosis and survival time of MCL. SOX11 cDNA is more sensitive than conventional methods for the detection of minimal residual disease (MRD). With further research of SOX11, it will become an important basis of diagnosis and prognosis in MCL, and can provide a new method for targeted therapy of MCL.
ABSTRACT
Objective Discuss the effect of the Rhythmic auditory stimulation with treadmill training on freezing of gait in patients with parkinson's disease.Method 68 patients with parkinson's disease randomly divided into 2 groups.The control group uses conventional drug treatment in parkinson's disease patients according to the condition of patients.The comprehensive training group using body weight support treadmill training combined with rhythmic stimulation training for 8 weeks.The two group subjects were proceeded gait tests by freezing of gait questionnaire 、functional independence measure、five time sit to stand test、timed up and go test、berg balance Scale eight weeks after training.Result The comprehensive training group had a significant difference compared with the control group (P ＜ 0.01)in decreased freezing of gait questionnaire score,decreased functional independence measure score,decreased five time sit to stand test times,decreased timed up and go test score,increased berg balance Scale score.Conclusion Rhythmic auditory stimulation with treadmill training can improve freezing of gait function of patients with parkinson's disease.