Objective:To study the inhibitory effects and mechanism of Armeniacae Semen Amarum alcohol extract in melanogenesis in zebrafish embryos.Methods:48 HPF healthy zebrafish embryos were randomly divided into blank control group, arbutin group, high-, medium- and low-dosage groups. The blank control group was added with 6 ml embryo culture medium, the arbutin group was added with 8 000 μg/ml arbutin solution 6 ml, and the high-, medium- and low-dosage groups were added with 2 925, 1 950 and 1 300 μg/ml Armeniacae Semen Amarum alcohol extract, respectively. After 24 hours of intervention, the melanin area of zebrafish embryos was observed by stereomicroscope, and the inhibition rate of ethanol extract of bitter almond on tyrosinase (TYR) activity and melanin content in zebrafish embryos were detected by spectrophotometry; the antioxidant activity was evaluated by DPPH radical scavenging capacity, ABTS radical scavenging capacity and CAT activity. RT-qPCR was used to detect the mRNA levels of MITF, Tyr and TYRP2 in zebrafish embryos.Results:Compared with the arbutin group, the low-, medium-, and high-dosage groups showed an increase in melanin area ( P<0.01 or P<0.05), a decrease in TYR activity and melanin content inhibition rate ( P<0.01), and IC50 values for DPPH and ABTS free radical scavenging rates of 23.636 and 3.195 mg/ml, respectively. The CAT activity increased in the medium- and high-dosage groups ( P<0.05 or P<0.01), while the mRNA levels of TYR and MITF decreased in the low-, medium-, and high-dosage groups ( P<0.01). Conclusion:Armeniacae Semen Amarum alcohol extract can effectively inhibit the production of melanin in zebrafish embryos, and its mechanism may be closely related to TYR activity and MITF related signaling pathway.

OBJECTIVE To establish a liquid chromatography massspectrometry(LC-MS/MS)method for quantitatively determining the concentration of cepharanthine in rat plasma and tissue samples after intravenous injection of cepharanthine hydrochloride.METHODS ①The LC-MS/MS method was adopted.A Phenomenex C18(3.0 mm×50 mm,2.6 μm)column was employed with a mobile phase consisting of 0.05%formic acid-2 mmol·L-1 ammonium acetate-water solution and 0.1%formic acid-acetonitrile solution under gradient elution at a flow rate of 0.6 mL·min-1.The determination was performed using positive ion multiple reaction monitoring mode assays:cepharanthine(m/z:607.3→365.1)and buspirone(IS)(m/z:386.4→122.2).② Blood samples were collected from 6 SD rats at different time points following a single iv administration of cepharanthine to determine the concentration of the drug.The main pharmacokinetic parameters were calculated using a non-compartmental model.③72 SD rats were subjected to tissue distribution experiments after a single and multiple iv administra-tion of cepharanthine,and tissue samples were collected at six different time points(n=6)for the quanti-fication of drug concentrations.④ The whole blood plasma distribution ratio(Rb/p)of cepharanthine hydrochloride(7.5 mg·kg-1)in 3 SD rats was determined 2 h after iv administration.⑤The protein binding of cepharanthine to rat plasma and lung tissue homogenates was determined by equilibrium dialysis before the concentration of the free drug within the lungs was calculated.RESULTS ① An LC-MS/MS method for quantitatively determining cepharanthine in rat plasma and tissue homogenates was devel-oped,which demonstrated an excellent linear relationship(r2>0.999)within the concentration range of 2 to 1000 μg·L-1,with a lower limit of quantification at 2 μg·L-1.The obtained results met all the require-ments for accurate quantitative detection.②The main pharmacokinetic parameters of cepharanthine in rats following a single iv administration were as follows:C0=(686.91±238.43)μg·L-1,t1/2=(29.70±6.29)h,Vz=(62.70±7.93)L·kg-1,Vss=(62.55±11.28)L·kg-1,CL=(1.50±0.23)L·h-1·kg-1 and AUC(0-t)=(4.52±0.61)h·mg·L-1.③ Concentrations in tissues exceeded those in plasma after both a single and multiple iv administration,with the highest levels in the lung.The values of AUC(0-t)in lungs were(2 547.35±156.56)and(4 481.35±479.21)h·mg·L-1 after a single and multiple iv administration,respectively.④ The content of cepharanthine in blood cells was higher than that in plasma,and Rb/p was 3.5±0.8.⑤ After correction by the protein-binding rate,the minimum concentration of free drugs in the lungs(95.04 μg·L-1)exceeded the reported antiviral activity threshold against coronaviruses(EC50=60.67 μg·L-1).CONCLUSION An LC-MS/MS method has been established to rapidly and sensitively determine the concentration of cepharanthine in rat plasma and tissues.Following intravenous administration of ceph-aranthine hydrochloride,the pulmonary exposure level of the drug is significantly higher in plasma and other tissues,providing data for evaluating its in vivo pharmacological activities.

Objective:To analyze the prevalence and risk factors of blindness and moderate to severe visual impairment among Han and Kazakh residents aged 50 years and older in Tacheng City, Xinjiang Uygur Autonomous Region.Methods:This study is a cross-sectional survey conducted using cluster random sampling from October 2015 to June 2018 in Emin County, Tacheng City, Xinjiang Uygur Autonomous Region. The study included individuals aged 50 years and older to survey blindness and moderate to severe visual impairment. Ophthalmological examinations combined with questionnaires were conducted to gather basic information. The data collected from the questionnaires included general demographic information and health conditions. The results of the eye examinations were used to diagnose a total of 12 risk factors including cataracts, glaucoma, pterygium, suspected glaucoma, glaucoma, and refractive errors. These risk factors were analyzed in relation to blindness and moderate to severe visual impairment. Univariate analysis was conducted first, followed by logistic regression to identify the significant factors.Results:A total of 2 114 patients were included in the final analysis, among which the prevalence of moderate to severe visual impairment was 18.54% (392/2 114), and the prevalence of blindness was 2.74% (58/2 114). Univariate analysis showed that blindness and moderate to severe visual impairment were associated with age ( χ2 = 32.97, P < 0.05), hypertension ( χ2 = 3.48, P < 0.05), age-related cataract ( χ2 = 17.43, P < 0.05), glaucoma ( χ2 = 3.90, P < 0.05), macular degeneration ( χ2 = 16.04, P < 0.05), diabetes ( χ2 = 3.09, P < 0.05), pterygium ( χ2 = 2.57, P < 0.05), and fundus arteriosclerosis ( χ2 = 2.31, P < 0.05). Multivariate logistic regression analysis indicated that moderate to severe visual impairment was correlated with age (50 to < 60 years: OR = 2.91, 95% CI: 0.44-13.45; 60 to < 70 years: OR = 3.52, 95% CI: 0.73-8.77; 70 to < 80 years: OR = 4.31, 95% CI: 0.85-8.96), ethnicity ( OR = 4.45, 95% CI: 0.56-5.95), sex ( OR = 0.47, 95% CI: 0.34-0.64), age-related cataract ( OR = 1.67, 95% CI: 1.05-2.65), glaucoma ( OR = 2.97, 95% CI: 1.67-5.30), and coronary heart disease ( OR = 2.56, P < 0.05). Blindness was correlated with age (70-79 years: OR = 1.54, 95% CI: 1.12-2.11), sex ( OR = 0.67, 95% CI: 0.34-0.64), glaucoma ( OR = 1.65, 95% CI: 0.42-6.49), diabetes ( OR = 2.05, 95% CI: 1.35-3.09), and coronary heart disease ( OR = 1.92, 95% CI: 1.07-3.43). Among these, age (70-79 years), glaucoma, diabetes, and coronary heart disease were identified as risk factors for blindness, while sex was observed as a protective factor against blindness in this region. Based on univariate and multivariate analyses as well as clinical practice, it was concluded that age (50 to < 60 years: OR = 4.42, 95% CI: 1.31-14.92; 60 to < 70 years: OR = 4.49, 95% CI: 1.70-11.84; 70 to < 80 years: OR = 3.19, 95% CI: 1.29-7.87), age-related cataract ( OR = 1.67, 95% CI: 1.05-2.65), and glaucoma ( OR = 2.97, 95% CI: 1.67-5.30) were identified as significant risk factors for moderate to severe visual impairment. Glaucoma ( OR = 1.65, 95% CI: 0.42-6.49) and diabetes ( OR = 2.05, 95% CI: 1.35-3.09) were identified as the main risk factors for blindness in this region (both P < 0.05). Conclusions:In Tacheng City, Xinjiang Uygur Autonomous Region, the prevalence rates of moderate to severe visual impairment and blindness among Han and Kazakh residents are relatively high. Age-related cataracts and glaucoma are the primary causes, while age and diabetes are the main risk factors.

Objective:To analyze the prevalence and risk factors of blindness and moderate to severe visual impairment among Han and Kazakh residents aged 50 years and older in Tacheng City, Xinjiang Uygur Autonomous Region.Methods:This study is a cross-sectional survey conducted using cluster random sampling from October 2015 to June 2018 in Emin County, Tacheng City, Xinjiang Uygur Autonomous Region. The study included individuals aged 50 years and older to survey blindness and moderate to severe visual impairment. Ophthalmological examinations combined with questionnaires were conducted to gather basic information. The data collected from the questionnaires included general demographic information and health conditions. The results of the eye examinations were used to diagnose a total of 12 risk factors including cataracts, glaucoma, pterygium, suspected glaucoma, glaucoma, and refractive errors. These risk factors were analyzed in relation to blindness and moderate to severe visual impairment. Univariate analysis was conducted first, followed by logistic regression to identify the significant factors.Results:A total of 2 114 patients were included in the final analysis, among which the prevalence of moderate to severe visual impairment was 18.54% (392/2 114), and the prevalence of blindness was 2.74% (58/2 114). Univariate analysis showed that blindness and moderate to severe visual impairment were associated with age ( χ2 = 32.97, P < 0.05), hypertension ( χ2 = 3.48, P < 0.05), age-related cataract ( χ2 = 17.43, P < 0.05), glaucoma ( χ2 = 3.90, P < 0.05), macular degeneration ( χ2 = 16.04, P < 0.05), diabetes ( χ2 = 3.09, P < 0.05), pterygium ( χ2 = 2.57, P < 0.05), and fundus arteriosclerosis ( χ2 = 2.31, P < 0.05). Multivariate logistic regression analysis indicated that moderate to severe visual impairment was correlated with age (50 to < 60 years: OR = 2.91, 95% CI: 0.44-13.45; 60 to < 70 years: OR = 3.52, 95% CI: 0.73-8.77; 70 to < 80 years: OR = 4.31, 95% CI: 0.85-8.96), ethnicity ( OR = 4.45, 95% CI: 0.56-5.95), sex ( OR = 0.47, 95% CI: 0.34-0.64), age-related cataract ( OR = 1.67, 95% CI: 1.05-2.65), glaucoma ( OR = 2.97, 95% CI: 1.67-5.30), and coronary heart disease ( OR = 2.56, P < 0.05). Blindness was correlated with age (70-79 years: OR = 1.54, 95% CI: 1.12-2.11), sex ( OR = 0.67, 95% CI: 0.34-0.64), glaucoma ( OR = 1.65, 95% CI: 0.42-6.49), diabetes ( OR = 2.05, 95% CI: 1.35-3.09), and coronary heart disease ( OR = 1.92, 95% CI: 1.07-3.43). Among these, age (70-79 years), glaucoma, diabetes, and coronary heart disease were identified as risk factors for blindness, while sex was observed as a protective factor against blindness in this region. Based on univariate and multivariate analyses as well as clinical practice, it was concluded that age (50 to < 60 years: OR = 4.42, 95% CI: 1.31-14.92; 60 to < 70 years: OR = 4.49, 95% CI: 1.70-11.84; 70 to < 80 years: OR = 3.19, 95% CI: 1.29-7.87), age-related cataract ( OR = 1.67, 95% CI: 1.05-2.65), and glaucoma ( OR = 2.97, 95% CI: 1.67-5.30) were identified as significant risk factors for moderate to severe visual impairment. Glaucoma ( OR = 1.65, 95% CI: 0.42-6.49) and diabetes ( OR = 2.05, 95% CI: 1.35-3.09) were identified as the main risk factors for blindness in this region (both P < 0.05). Conclusions:In Tacheng City, Xinjiang Uygur Autonomous Region, the prevalence rates of moderate to severe visual impairment and blindness among Han and Kazakh residents are relatively high. Age-related cataracts and glaucoma are the primary causes, while age and diabetes are the main risk factors.

OBJECTIVE To establish a liquid chromatography massspectrometry(LC-MS/MS)method for quantitatively determining the concentration of cepharanthine in rat plasma and tissue samples after intravenous injection of cepharanthine hydrochloride.METHODS ①The LC-MS/MS method was adopted.A Phenomenex C18(3.0 mm×50 mm,2.6 μm)column was employed with a mobile phase consisting of 0.05%formic acid-2 mmol·L-1 ammonium acetate-water solution and 0.1%formic acid-acetonitrile solution under gradient elution at a flow rate of 0.6 mL·min-1.The determination was performed using positive ion multiple reaction monitoring mode assays:cepharanthine(m/z:607.3→365.1)and buspirone(IS)(m/z:386.4→122.2).② Blood samples were collected from 6 SD rats at different time points following a single iv administration of cepharanthine to determine the concentration of the drug.The main pharmacokinetic parameters were calculated using a non-compartmental model.③72 SD rats were subjected to tissue distribution experiments after a single and multiple iv administra-tion of cepharanthine,and tissue samples were collected at six different time points(n=6)for the quanti-fication of drug concentrations.④ The whole blood plasma distribution ratio(Rb/p)of cepharanthine hydrochloride(7.5 mg·kg-1)in 3 SD rats was determined 2 h after iv administration.⑤The protein binding of cepharanthine to rat plasma and lung tissue homogenates was determined by equilibrium dialysis before the concentration of the free drug within the lungs was calculated.RESULTS ① An LC-MS/MS method for quantitatively determining cepharanthine in rat plasma and tissue homogenates was devel-oped,which demonstrated an excellent linear relationship(r2>0.999)within the concentration range of 2 to 1000 μg·L-1,with a lower limit of quantification at 2 μg·L-1.The obtained results met all the require-ments for accurate quantitative detection.②The main pharmacokinetic parameters of cepharanthine in rats following a single iv administration were as follows:C0=(686.91±238.43)μg·L-1,t1/2=(29.70±6.29)h,Vz=(62.70±7.93)L·kg-1,Vss=(62.55±11.28)L·kg-1,CL=(1.50±0.23)L·h-1·kg-1 and AUC(0-t)=(4.52±0.61)h·mg·L-1.③ Concentrations in tissues exceeded those in plasma after both a single and multiple iv administration,with the highest levels in the lung.The values of AUC(0-t)in lungs were(2 547.35±156.56)and(4 481.35±479.21)h·mg·L-1 after a single and multiple iv administration,respectively.④ The content of cepharanthine in blood cells was higher than that in plasma,and Rb/p was 3.5±0.8.⑤ After correction by the protein-binding rate,the minimum concentration of free drugs in the lungs(95.04 μg·L-1)exceeded the reported antiviral activity threshold against coronaviruses(EC50=60.67 μg·L-1).CONCLUSION An LC-MS/MS method has been established to rapidly and sensitively determine the concentration of cepharanthine in rat plasma and tissues.Following intravenous administration of ceph-aranthine hydrochloride,the pulmonary exposure level of the drug is significantly higher in plasma and other tissues,providing data for evaluating its in vivo pharmacological activities.

The extraordinary advantages associated with mRNA vaccines, including their high efficiency, relatively low severity of side effects, and ease of manufacture, have enabled them to be a promising immunotherapy approach against various infectious diseases and cancers. Nevertheless, most mRNA delivery carriers have many disadvantages, such as high toxicity, poor biocompatibility, and low efficiency in vivo, which have hindered the widespread use of mRNA vaccines. To further characterize and solve these problems and develop a new type of safe and efficient mRNA delivery carrier, a negatively charged SA@DOTAP-mRNA nanovaccine was prepared in this study by coating DOTAP-mRNA with the natural anionic polymer sodium alginate (SA). Intriguingly, the transfection efficiency of SA@DOTAP-mRNA was significantly higher than that of DOTAP-mRNA, which was not due to the increase in cellular uptake but was associated with changes in the endocytosis pathway and the strong lysosome escape ability of SA@DOTAP-mRNA. In addition, we found that SA significantly increased the expression of LUC-mRNA in mice and achieved certain spleen targeting. Finally, we confirmed that SA@DOTAP-mRNA had a stronger antigen-presenting ability in E. G7-OVA tumor-bearing mice, dramatically inducing the proliferation of OVA-specific CLTs and ameliorating the antitumor effect. Therefore, we firmly believe that the coating strategy applied to cationic liposome/mRNA complexes is of potential research value in the field of mRNA delivery and has promising clinical application prospects.

Objective:To investigate the effect of ionizing radiation on the N 6-methyladenine (m 6A) modification profile of circular RNA (circRNA) in mouse bone marrow cells and provide scientific basis for revealing the relationship between RNA epigenetic modification and hematopoietic radiation injury. Methods:A total of twenty four C57BL/6 J mice were randomly divided into two groups: the healthy control group ( n=12), and ionizing radiation group ( n=12) irradiated in total body with 4 Gy of 137Cs γ-rays. At 5 min after irradiation, mice were killed and bone marrow cells were collected from the femur. Total RNAs were extracted and the changes in circRNA m6A modification profiles were investigated by RNA immunoprecipitation-high-throughput sequencing (MeRIP-Seq) technology and bioinformatics analysis. The representative alterations of m 6A peaks were validated by MeRIP-PCR assay. Results:325 and 455 m 6A sites were identified on circRNAs in the healthy control group and ionizing radiation group (178 common sites, 147 specific sites in the healthy control group and 277 specific sites in ionizing radiation group), respectively. 1 275 and 1 017 deriving genes of m 6A-circRNAs were identified in the healthy control group and ionizing radiation group (767 common genes, 508 specific genes in the healthy control group and 250 specific genes in ionizing radiation group), respectively. Compared with the control healthy group, 414 (178) m 6A peaks was significantly up- (down-) regulated in the ionizing radiation group( P < 10 -10; fold-change cut-off > 5). Moreover, Gene Ontology (GO) assay revealed that the deriving genes of circRNAs with differentially methylated m 6A sites between two groups involves various functions including chromatin regulation, ciliary transition fiber and poly (A)-specific ribonuclease activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) assay revealed that the deriving genes of circRNAs with differentially methylated m 6A sites between two groups included numerous pathways such as platelet activation, Fc γ R-mediated phagocytosis and B cell receptor signaling pathway. Conclusions:Ionizing radiation triggers rapid alterations in the m 6A modification profile of circRNA in mouse bone marrow cells. The deriving genes of differentially methylated circRNAs are associated with a variety of functions and signaling pathways of hematopoietic radiobiology.

Intestinal schistosomiasis is caused by infestation with schistosome, which is easy to be confused with inflammatory bowel disease, especially Crohn′s disease or malignant tumor. Such a case that was misdiagnosed as Crohn′s disease in Zhongnan hospital is described. Schistosome eggs, granuloma and eosinophil infiltration observed in resected specimens can help reaching the correct diagnosis.

Intestinal schistosomiasis is caused by infestation with schistosome, which is easy to be confused with inflammatory bowel disease, especially Crohn′s disease or malignant tumor. Such a case that was misdiagnosed as Crohn′s disease in Zhongnan hospital is described. Schistosome eggs, granuloma and eosinophil infiltration observed in resected specimens can help reaching the correct diagnosis.

Objective To detect the protein expression change in the proliferation of human retinal microvascular endothelial cells (hRMECs) stimulated with 4-Hydroxynonenal (4-HNE).Methods hRMECs were in a logarithmic growth phase, and then were separated into 4-HNE-stimulated group and negative control group. The concentration of 4-HNE included 5, 10, 20 and 50 μmol/L in 4-HNE-stimulated group, while the negative control group was added in the same volume of ethanol (the solvent of 4-HNE). Then the cells were stimulated with 4-HNE for 24 hours following by CCK-8 kits incubating for 4 hours to detect absorbance. It was found that 10 μmol/L 4-HNE had the most obvious effect on the proliferation of hRMECs. Therefore, the cellular proteins from 10 μmol/L 4-HNE-stimulated group and negative control group were acquired and prepared by FASP sample preparation method. Data independent acquisition was used for data acquisition, and the GO analysis and pathway enrichment were performed for analysis of differentially expressed proteins. Results CCK-8 kits detection results showed that theA value of the 10 and 20 μmol/L 4-HNE-stimulated groups were significantly higher than negative control group and 5 μmol/L 4-HNE-stimulated group (F=25.42, P<0.01), while there were no differences between 10 and 20 μmol/L 4-HNE-stimulated groups, and theA value of 50 μmol/L 4-HNE-stimulated groups was lower than negative control. A total of 2710 quantifiable proteins were identified by peoteomics,and 118 proteins were differentially expressed (fold change>1.5,P<0.05). Seventy-two proteins were up-regulated after 4-HNE stimulation, whereas 46 proteins were down-regulated. Particularly, the expressions of Heme oxygenase-1, Sulfoxdoxin-1, Heat shock protein A1B, Thioredoxin reductase-1, Glutathione reductase, ATPase and prothrombin were increased when cells were added in 4-HNE, whereas the expressions of apolipoprotein A1 and programmed cell death protein 4 were decreased. These differentially expressed proteins were mainly involved in the biological processes such as oxidative stress, cell detoxification, and ATPase-coupled membrane transport.Conclusion After stimulated with 4-HNE, the oxidative stress, cell detoxification, and ATPase-coupled membrane transport protein expression may change in hRMECs in order to regulate oxidative stress and growth situation.