Effect Mechanism of Modified Baitouwengtang Treating Colorectal Cancer by Regulating CSF1R/STING/TBK1 Signaling to Polarize Phenotype of Tumor-associated Macrophages （TAMs） / 中国实验方剂学杂志

ObjectiveThis study aims to investigate the effect of modified Baitouwengtang （MBTWD） on tumor growth and the number of tumor-associated macrophages （TAMs） in tumor tissue of MC38 cell tumor-bearing mice with colorectal cancer and explores whether MBTWD mediates the remodeling of TAM phenotype to play an immunologically antitumor effect. MethodFirstly， The C57BL/6 mouse tumor model grafted subcutaneously was established， and then model mice were classified into a model group， positive control group（3 mg·kg-1）， and MBTWD groups with high and low dosages（23.43、46.86 g·kg-1）， with 10 mice in each group. In addition， 10 healthy mice were set as the blank group， and the changes in body weight， tumor volume， and survival status of mice in each group were observed. Tumor tissue， spleen， and peripheral blood were collected to calculate the tumor volume change， tumor inhibition rate， and spleen mass. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of tumor tissue， and an immunofluorescence assay was used to detect the expression levels of CD4， CD8， and CD206 in tumor tissues of tumor-bearing mice. The secretion levels of transforming growth factor （TGF）-β， interleukin （IL）-6， and chemokine （C-C Motif） ligand 2 （CCL2） in peripheral serum were measured by using enzyme-linked immunosorbent assay （ELISA）. Secondly， a co-culture model induced by IL-4 in vitro of MC38 cells and murine monocytic macrophage RAW264.7 cells was established. Cell proliferation and activity assay （CCK-8） was used to detect the inhibitory effect of MBTWD containing serum on cell proliferation. A transwell experiment was used to detect the effect of IL-4-induced M2 macrophages on the invasion of MC38 cells. Flow cytometry was used to detect the expression of CD86 on the membrane of M2 macrophages induced by IL-4 with MBTWD containing serum. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the effect of MBTWD containing serum on the mRNA expression levels of M1 macrophage-related polarization factors CD86， nitric oxide synthase （iNOS）， and IL-12， as well as M2 macrophage-related polarization factors CD206， CD163， and IL-10 after co-cultivation. Finally， the protein expression levels of colony-stimulating factor 1 receptor （CSF1R）， stimulator of interferon genes （STING）， and TANK binding kinase 1 （TBK1） in tumor tissues of tumor-bearing mice were detected by Western blot. ResultIn vivo experimental results show that compared with the model group， the MBTWD can significantly inhibit the tumor growth of tumor-bearing mice. Immunofluorescence experiments show that the MBTWD can increase the number of CD8+ T cell infiltration in tumor tissue of tumor-bearing mice， reduce the number of CD206+ TAMs infiltration， and down-regulate the secretion levels of cytokines IL-6， TGF-β， and CCL2 in peripheral blood of tumor-bearing mice. The results of in vitro experiments show that the MBTWD containing serum has no obvious inhibitory effect on cell proliferation， but the cell supernatant after co-cultivation with RAW264.7 cells can inhibit the proliferation activity of MC38 cells， and the invasion ability of MC38 cells is enhanced by IL-4-induced M2 macrophages. However， this effect can be inhibited in a concentration-dependent manner by the MBTWD containing serum. At the same time， the results of Real-time PCR show that the MBTWD containing serum can up-regulate the mRNA expression levels of M1 macrophage-related polarization factors CD86， iNOS， and IL-12 and down-regulate those of M2 macrophage-related polarization factors CD206， CD163， and IL-10. Flow cytometry results also confirm that the MBTWD containing serum can increase the number of repolarized CD86+ M1 macrophages， indicating that MBTWD can induce M2 macrophages to repolarized M1 macrophages to play an anti-tumor growth role. Finally， Western blot results show that MBTWD can down-regulate the expression of CSF1R protein and up-regulate that of STING and TBK1 proteins in tumor tissue of tumor-bearing mice. ConclusionMBTWD can down-regulate the infiltration number of CD206+ TAMs and increase the infiltration of CD8+ T cells， thereby playing an immunologically antitumor effect on the growth inhibition of colorectal cancer， which may be related to regulating CSF1R signaling and then activating STING/TBK1 signaling pathway to induce phenotypic remodeling of TAMs.

Progress of Different Programmed Cell Death Pathways in Kidney Cancer / 肿瘤防治研究

Programmed cell death (PCD) is a genetically determined, active and orderly cell death in the organism, and it affects the evolution of the organism, maintenance of its homeostasis, and development of several tissues and organs. The abnormal regulation of this process is closely related to various human diseases, including cancer. The identified pathways of PCD include apoptosis, autophagy, necroptosis, pyroptosis, and ferroptosis, which can be activated when cells are stimulated by various internal and external environmental factors. These pathways can induce cell death or maintain cell survival in kidney cancer cells under the regulation of various signaling molecules, thus affecting tumor progression or therapeutic efficacy. In this paper, the role of these PCD pathways in the development of kidney cancer was reviewed in light of recent research advances to provide new directions for the in-depth study of the pathogenesis of kidney cancer and the development of targeted antitumor drugs.

Pyridine pigments from functional Monascus rice / 药学学报

The trace chemical components in functional Monascus rice were studied to explore the potential bioactive substances. MCI column, Sephadex LH-20 gel, and preparative liquid chromatography were used to purified the ethyl acetate extract from functional Monascus rice. Two novel pyridine Monascus pigments were isolated and identified, named monascopyridine G (1) and monascopyridine H (2), respectively based on extensive mass spectrometry (MS), infrared radiation (IR), and nuclear magnetic resonance (NMR) analysis. The molecular docking experiments between compounds 1 and 2 and peroxisome proliferators-activated receptor-gamma (PPARγ) showed that they exhibited obvious binding force with the receptor protein. Besides, the biosynthetic pathways of the two compounds were proposed, which provide a valuable reference for the selective production of these potential bioactive substances.

Mid-term results of endovascular intervention for transplant renal artery stenosis / 中华普通外科杂志

Objective:To evaluate the mid-term results of endovascular treatment for transplant renal artery stenosis (TRAS).Methods:The clinical and follow-up data of TRAS patients undergoing endovascular treatment at the First Affiliated Hospital of Zhengzhou University from Jan 2014 to Jan 2021 were retrospectively analyzed.Results:A total of 2 230 patients underwent kidney transplantation, 78 cases(3.6%) developed TRAS, among those 27 patients received endovascular treatment and followed-up from 12 to 80 months(mean 36 months). Thirteen patients (48.1%) underwent renal graft angiography and balloon dilatation, of which 2 patients underwent stent placement, 14 patients (51.9%) underwent renal graft angiography with balloon dilatation and stenting. The serum creatinine 2 weeks postoperatively and 12 months postoperatively were 127.6 μmol/L (47-220 μmol/L) and 103.4 μmol/L (63-166 μmol/L), respectively, significantly lower than the preoperative 217.1 μmol/L (98-541 μmol/L), ( P<0.05). Glomerular filtration rate (GFR) before surgery was 8.3-105.3 ml/min, 2 weeks and 12 months after surgery compared to 24.6-132.2 ml/min and 47.3-113.9 ml/min( P<0.05). The preoperative peak systolic velocity (PSV) of the transplanted renal artery during the systolic phase was 234 cm/s (75-457 cm/s), compared to 129 cm/s (52-290 cm/s) ( P<0.05) 2 weeks and 118 cm/s (57-300 cm/s) 12 months postoperatively ( P<0.05). During the follow-up period, 2 patients (7.4%) died of multiple organ failure. Conclusions:TRAS is the most common vascular complication after kidney transplantation. Endovascular treatment has a high success rate and low complication rate.

Chloroplast genome phylogeny and codon preference of Docynia longiunguis / 生物工程学报

Docynia longiunguis is a plant uniquely present in China and is of high edible and medicinal value. The analysis of its chloroplast genome will help clarify the phylogenetic relationship among Docynia and facilitate the development and utilization of D. longiunguis resources. Based on the alignment of chloroplast genome sequences of related species, the phylogeny and codon preference were analyzed. The total length of D. longiunguis chloroplast genome sequence was 158 914 bp (GenBank accession number is MW367027), with an average GC content of 36.7%. The length of the large single-copy (LSC), the small single-copy (SSC), and inverted repeats (IRs) are 87 020 bp, 19 156 bp, and 26 369 bp, respectively. A total of 102 functional genes were annotated, including 72 protein-coding genes, 26 tRNA genes, and 4 rRNA genes. The best model for constructing phylogenetic tree was TVM+F+R2. D. longiunguis and Docynia indica were clustered into a single group, while Docynia and Malus were clustered into a single group. Comparison of the chloroplast genome sequences of D. longiunguis and its five related species revealed that trnY (GUA)-psbD, ndhC-trnV (UAC), accD-psaI, psbZ-trnfM (CAU), ndhF-trnL gene regions varied greatly. The nucleic acid diversity analysis showed that there were 11 high variation areas with nucleotide variability > 0.01, all were located in the LSC and SSC regions. Except for D. longiunguis, the trnH genes in other sequences were located at the IRs/LSC junction and did not cross the boundary. Codon preference analysis showed that D. longiunguis chloroplast genome has the largest number of isoleucine (Ile) codons, up to 1 205. D. longiunguis has the closest genetic relationship with Malus baccata, Malus sieboldii, Malus hupehensis and Chaenomeles sinensis. Its chloroplast genome codon prefers to end with A/T. The chloroplast genome of D. longiunguis and other Rosaceae chloroplast genomes showed great differences in gene distribution in four boundary regions, while relatively small differences from the chloroplast genomes of Docynia delavayi and D. indica of the same genus were observed. The genome annotation, phylogenetic analysis and sequence alignment of chloroplast genome of D. longiunguis may facilitate the identification, development and utilization of this species.

Genomic Epidemiology of Imported Cases of COVID-19 in Guangdong Province, China, October 2020 - May 2021 / 生物医学与环境科学（英文）

Objective@#The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been engendering enormous hazards to the world. We obtained the complete genome sequences of SARS-CoV-2 from imported cases admitted to the Guangzhou Eighth People's Hospital, which was appointed by the Guangdong provincial government to treat coronavirus disease 2019 (COVID-19). The SARS-CoV-2 diversity was analyzed, and the mutation characteristics, time, and regional trend of variant emergence were evaluated.@*Methods@#In total, 177 throat swab samples were obtained from COVID-19 patients (from October 2020 to May 2021). High-throughput sequencing technology was used to detect the viral sequences of patients infected with SARS-CoV-2. Phylogenetic and molecular evolutionary analyses were used to evaluate the mutation characteristics and the time and regional trends of variants.@*Results@#We observed that the imported cases mainly occurred after January 2021, peaking in May 2021, with the highest proportion observed from cases originating from the United States. The main lineages were found in Europe, Africa, and North America, and B.1.1.7 and B.1.351 were the two major sublineages. Sublineage B.1.618 was the Asian lineage (Indian) found in this study, and B.1.1.228 was not included in the lineage list of the Pangolin web. A reasonably high homology was observed among all samples. The total frequency of mutations showed that the open reading frame 1a (ORF1a) protein had the highest mutation density at the nucleotide level, and the D614G mutation in the spike protein was the commonest at the amino acid level. Most importantly, we identified some amino acid mutations in positions S, ORF7b, and ORF9b, and they have neither been reported on the Global Initiative of Sharing All Influenza Data nor published in PubMed among all missense mutations.@*Conclusion@#These results suggested the diversity of lineages and sublineages and the high homology at the amino acid level among imported cases infected with SARS-CoV-2 in Guangdong Province, China.

Kinetic Characteristics of Neutralizing Antibody Responses Vary among Patients with COVID-19 / 生物医学与环境科学（英文）

Objective@#The coronavirus disease 2019 (COVID-19) pandemic continues to present a major challenge to public health. Vaccine development requires an understanding of the kinetics of neutralizing antibody (NAb) responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).@*Methods@#In total, 605 serum samples from 125 COVID-19 patients (from January 1 to March 14, 2020) varying in age, sex, severity of symptoms, and presence of underlying diseases were collected, and antibody titers were measured using a micro-neutralization assay with wild-type SARS-CoV-2.@*Results@#NAbs were detectable approximately 10 days post-onset (dpo) of symptoms and peaked at approximately 20 dpo. The NAb levels were slightly higher in young males and severe cases, while no significant difference was observed for the other classifications. In follow-up cases, the NAb titer had increased or stabilized in 18 cases, whereas it had decreased in 26 cases, and in one case NAbs were undetectable at the end of our observation. Although a decreasing trend in NAb titer was observed in many cases, the NAb level was generally still protective.@*Conclusion@#We demonstrated that NAb levels vary among all categories of COVID-19 patients. Long-term studies are needed to determine the longevity and protective efficiency of NAbs induced by SARS-CoV-2.

Potential effect of EGCG on the anti-tumor efficacy of metformin in melanoma cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Metformin, a first-line drug for type 2 diabetes mellitus, has been recognized as a potential anti-tumor agent in recent years. Epigallocatechin-3-gallate (EGCG), as the dominant catechin in green tea, is another promising adjuvant agent for tumor prevention. In the present work, the potential effect of EGCG on the anti-tumor efficacy of metformin in a mouse melanoma cell line (B16F10) was investigated. Results indicated that EGCG and metformin exhibited a synergistic effect on cell viability, migration, and proliferation, as well as signal transducer and activator of transcription 3/nuclear factor-κB (STAT3/NF-κB) pathway signaling and the production of inflammation cytokines. Meanwhile, the combination showed an antagonistic effect on cell apoptosis and oxidative stress levels. The combination of EGCG and metformin also differentially affected the nucleus (synergism) and cytoplasm (antagonism) of B16F10 cells. Our findings provide new insight into the potential effects of EGCG on the anti-tumor efficacy of metformin in melanoma cells.

Expression of bone marrow stromal antigen 2 gene in patients with glioblastoma and its correlation with non-CpG island DNA methylation and prognosis / 肿瘤研究与临床

Objective:To investigate the expression level of bone marrow stromal antigen 2 (BST2) in patients with glioblastoma (GBM) gene and its correlation with DNA methylation level and prognosis.Methods:The datasets of GBM samples from the Cancer Genome Atlas (TCGA) database (35 cases), GSE22891 cohort (50 cases) within Gene Expression Omnibus (GEO) database, and China Glioma Genome Atlas (CGGA) database (105 cases), and non-tumor brains (NTB) (10 cases in TCGA database, 6 cases in GSE22891 cohort, 5 cases in CGGA database were used to make comparisons of gene expression level; 25 cases in GSE63347 cohort, 4 cases in GSE22891 cohort, 8 cases in CGGA database were used to make comparisons of methylation data). Based on gene expression chip and DNA methylation microarray data from public GBM databases, the expression level of BST2 gene in GBM and the association with non-CpG island DNA methylation, GBM molecular subtypes [CpG island methylation phenotype (G-CIMP) and non-G-CIMP proneural, neural, classical, mesenchymal], overall survival (OS) time and functional gene expression profiles were obtained by using intra-group comparison of BST2 gene expression level between GBM and NTB, survival analysis, and bioinformatic analysis.Results:Compared with NTB samples, BST2 mRNA was highly expressed in GBM (mRNA expression data were based on Z-score standardization) (TCGA database vs. GSE63347 cohort: -0.97±1.14 vs. -2.32±0.21, t = 3.74, P < 0.05; GSE22891 cohort: 9.03±1.28 vs. 7.18±0.22, t = 3.42, P < 0.05; CGGA database: -0.43±1.11 vs. -0.62±0.35, t = 2.09, P < 0.05). In TCGA database, BST2 mRNA was highly expressed in different tumor molecular subgroups (G-CIMP: -1.96±0.94; non-G-CIMP proneural: -1.74±0.88; neural: -0.83±0.98; classical: 0.71±1.18; mesenchymal: -0.55±1.01), which were compared with NTB samples (-2.32±0.21), and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in the expressions of BST2 mRNA in the neural, classical, mesenchymal tumors (all P > 0.05), but the expression of BST2 mRNA in above three groups was higher than that in G-CIMP group and non-G-CIMP proneural group (all P < 0.05). Correlation analysis showed that non-CpG island DNA methylation level of BST2 was negatively correlated with the expression of its mRNA (TCGA database: r = -0.30, P < 0.05; GSE22891 cohort: r = -0.54, P < 0.05; CGGA database: r = -0.29, P > 0.05). Survival analysis showed that BST2 mRNA expression level of non-G-CIMP and non-proneural patients was negatively associated with OS ( HR = 1.18, 95% CI 1.00-1.39, P < 0.05); among those tumors with G-CIMP or proneural subtypes, BST2 mRNA expression was not associated with OS ( HR = 1.08, 95% CI 0.84-1.40, P > 0.05). Bioinformatic analysis showed that among non-G-CIMP and non-proneural samples of TCGA database, GBM samples with higher BST2 expression were enriched with functional gene sets related to negative regulation of immune responses and activation of nuclear factor κB (NF-κB) pathway. Conclusions:The upregulated expression of BST2 gene in GBM may be associated with non-CpG island DNA hypomethylation alteration. BST2 gene may become a potential prognostic biomarker for non-G-CIMP and non-proneural GBM.

Willingness for downward referral and its influencing factors among rural elderlies with hospital stay in Shandong province / 中华医院管理杂志

Objective@#To understand the referral status and the willingness for downward referral among rural elderlies with hospital stay experiences in the past year in Shandong province, and to explore its influencing factors on the willingness for downward referral.@*Methods@#Three prefecture-level cities in Shandong province were sampled by multi-stage stratified random sampling in August 2017. Questionnaire survey was conducted among 910 rural elderlies(over 60 years old)who had been hospitalized in the past year. The study included the basic information, the hospitalization experience, and perception of essential medicines system, ego-resiliency, the referral status and willingness for downward referral. Rank sum test, Chi-square test and t test were used for univariate analysis, and logistic regression was applied for influencing factors.@*Results@#Among 910 rural elderlies who had been hospitalized in the past year, 53(5.8%) were referred to other medical institutions in their recent hospitalization, and 597(65.6%)were willing to be referred downward. The main reason for their reluctance for downward referral was that the medical competency of primary medical institutions was low; and the main reason for willingness for downward referral was being close to home. The results showed that marital status, impression for the national essential medicines system and ego-resiliency were the influencing factors of their willingness to downward referral among rural elderlies who had been hospitalized in the past year(P<0.05).@*Conclusions@#The referral rate and willingness for downward referral among the rural elderlies in Shandong province are relatively low. We should further enhance the service capacity of primary medical institutions, strengthen the publicity and implementation of the national essential medicines system, pay attention to help with the elderlies′ negative emotions caused by diseases, and improve their ego-resiliency.

Utilization of inpatient health care and its influencing factors among elderlies in Shandong province / 中华医院管理杂志

Objective@#To investigate the utilization of inpatient health care among the elderlies in Shandong province, and to analyze the factors affecting the inpatient services utilization, so as to provide reference for the elderlies to utilize the inpatient services reasonably.@*Methods@#The survey was conducted in Shandong province in August 2017. Multi-stage stratified cluster random sampling method was used to select 7 070 residents aged 60 and above in 6 counties and districts of Shandong province as the objects of the survey. The survey included the basic family and personal information of the elderlies as well as the utilization of hospitalization services. Chi-square test and rank sum test were used for univariate analysis, and logistic regression was applied for influencing factors.@*Results@#The annual hospitalization rate of the elderlies in Shandong province was 18.1%, and 9.6% of those in need of hospitalization failed to enjoy the service. The annual hospitalization rate of the elderlies aged 80 years and over was 19.9%, and 5.5% of the patients in need had not been hospitalized. Among the inpatient institutions, the proportion of township health centers/community health service centers, county-level(district) medical institutions, prefecture-level medical institutions and provincial-level medical institutions was 29.2%, 29.1%, 37.7% and 1.4%, respectively.Factors influencing the utilization of hospitalization services for the elderlies included age, self-assessment of health, physical examination, chronic diseases, type of medical insurance and income level.@*Conclusions@#More attention should be paid to the hospitalization services for the elderlies aged 80 years and over. Effective measures should be taken to guide the elderlies to fully use primary medical resources. The prevention and control of chronic diseases should be strengthened to promote the rational use of inpatient health services among the elderlies. In addition, more attention should be paid to low-income elderlies to meet their hospitalization needs.

The effect of STC1 gene on radiosensitivity of human lung cancer A549 cell line / 中华放射肿瘤学杂志

Objective To investigate effect of stanniocalcin-1 (STC1) gene on the proliferation,apoptosis and radiotherapy sensitivity of non-small cell lung cancer.Methods The STC1 siRNA (STC 1-siRNA) and the non-interfering siRNA (negative control group) were transfected into the human lung cancer A549 cells by LipofectamineTM2000,and the blank control group was established.The expression level of STC1 protein was detected after transfection for 48 h by Western blotting.Clone forming test was adopted to detect the proliferation of A549 cells after STC1-siRNA and irradiation treatment.CCK8 assay was performed to detect the cell viability after treatment with STC1-siRNA and STC1-siRNA+8 Gy.The cell apoptosis was detected by flow cytometry.The expression levels of Ki67,Bax,STAT3 and p-STAT3 proteins were quantitatively measured by Western blotting.Results The expression level of STC1 protein in the A549 cells transfected with STC1-siRNA was significantly down-regulated than that in the blank control group (P＜ 0.05).Compared with the blank control group,the sensitization ratio was significantly enhanced after STC1-siRNA transfection.Compared with the blank control group,the cell viability and the expression levels of Ki67 and p-STAT3 protein were significantly decreased,whereas the apoptosis rate and the expression of Bax protein were significantly increased in the STC1-siRNA group.Compared with the STC1-siRNA group,the cell viability and the expression levels of Ki67 and p-STAT3 proteins were significantly decreased,whereas the cell apoptosis rate and the expression of Bax protein were remarkably increased in the STC1-siRNA+ 8 Gy group (all P＜0.05).Conclusion Inhibition of STC1 gene expression can enhance the radiotherapy sensitivity and down-regulate the STAT3 signaling pathway in non-small cell lung cancer.

Protective effects of 17β-estradiol on lens epithelial cells against H2O2-induced oxidative stress / 眼科新进展

Objective To investigate the protective effects of 17β-estradiol on human lens epithelial (HLE) cells in oxidative damage induced by H2 O2 and its involved mechanisms.Methods HLE cells cultured in vitro were collected and divided into 4 groups;cells in negative control group were cultured with normal medium,cells with 80％ fusion in n2O2 damage group was treated with 100 μmol · L-1 H2O2 for 12 h,cells in 17β-estradiol low dose group were incubated with 10 μmol · L-1 17β-estradiol for 24 h then subjected to 100 μmol · L-1 H2O2 for 12 h and ceEs in 17β-estradiol high dose group were cultured in 100 μmol · L-1 17β-estradiol for 24 h then treated with 100 μrnol · L-1 H2O2 for 12h.Next,cell viability was tested by CCK-8 colorimetric assay,while apoptotic rate was detected by flow cytometry,and intracellular reactive oxygen species (ROS) level was detected by H2 DCFDA fluorescence probe labeling method,as well as the contents of catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined by 721 D spectrophotometer.Results When compared with the negative control group,the survival rate of HLE ceils in the H2 O2 damage group was significantly decreased,and there was significantly different between both groups (P ＜0.05).Moreover,the survival rate of the 17β-estradiol low dose group (67.44％) and high dose group (78.52％) was obviously higher than that of the H2O2 damage group (59.34％),and the difference was statistically significant (P ＜ 0.05).After H2O2-induced injury,there was significant difference in the apoptotic rate of HLE cells between the H2O2 damage group (41.30 ±3.21)％ and the negative control group (1.67+0.32)％.In addition,the apoptotic rates of the 17β-estradiol low dose group and high dose group was (20.97 + 1.13)％ and (14.27 + 0.90)％ respectively,which was statistically different from the H2 O2 damage group (all P ＜ 0.05).H2 DCFDA fluorescent labeling test results showed that ROS fluorescence signal intensity gradually weakened after treated with 17β-estradiol.Besides,17β-estradiol significantly increased the expression of CAT、SOD and GSH-Px in HLE cells.Conclusion 17β-estradiol has obvious protective effects on HLE cells from the damage induced by H2O2.

Protective effects of 17β-estradiol on lens epithelial cells against H2O2-induced oxidative stress / 眼科新进展

Objective To investigate the protective effects of 17β-estradiol on human lens epithelial (HLE) cells in oxidative damage induced by H2 O2 and its involved mechanisms.Methods HLE cells cultured in vitro were collected and divided into 4 groups;cells in negative control group were cultured with normal medium,cells with 80％ fusion in n2O2 damage group was treated with 100 μmol · L-1 H2O2 for 12 h,cells in 17β-estradiol low dose group were incubated with 10 μmol · L-1 17β-estradiol for 24 h then subjected to 100 μmol · L-1 H2O2 for 12 h and ceEs in 17β-estradiol high dose group were cultured in 100 μmol · L-1 17β-estradiol for 24 h then treated with 100 μrnol · L-1 H2O2 for 12h.Next,cell viability was tested by CCK-8 colorimetric assay,while apoptotic rate was detected by flow cytometry,and intracellular reactive oxygen species (ROS) level was detected by H2 DCFDA fluorescence probe labeling method,as well as the contents of catalase (CAT),superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined by 721 D spectrophotometer.Results When compared with the negative control group,the survival rate of HLE ceils in the H2 O2 damage group was significantly decreased,and there was significantly different between both groups (P ＜0.05).Moreover,the survival rate of the 17β-estradiol low dose group (67.44％) and high dose group (78.52％) was obviously higher than that of the H2O2 damage group (59.34％),and the difference was statistically significant (P ＜ 0.05).After H2O2-induced injury,there was significant difference in the apoptotic rate of HLE cells between the H2O2 damage group (41.30 ±3.21)％ and the negative control group (1.67+0.32)％.In addition,the apoptotic rates of the 17β-estradiol low dose group and high dose group was (20.97 + 1.13)％ and (14.27 + 0.90)％ respectively,which was statistically different from the H2 O2 damage group (all P ＜ 0.05).H2 DCFDA fluorescent labeling test results showed that ROS fluorescence signal intensity gradually weakened after treated with 17β-estradiol.Besides,17β-estradiol significantly increased the expression of CAT、SOD and GSH-Px in HLE cells.Conclusion 17β-estradiol has obvious protective effects on HLE cells from the damage induced by H2O2.

Based on in vivo fluorescence imaging technology, to extablish a fluorescence modification method for amphipathic block polymers / 药学学报

Rhodamine B (RhB) was used to decorate an amphipathic block polymers (β-CD-[P(AA-co-MMA)-b-PVP]4) in this study. First, after activated by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, rhodamine B was marked with hydroxyethyl methacrylate (HEMA) through ester exchange reaction. Second, the labeled amphipathic block polymers (β-CD-[P(AA-(HEMA-RhB)-MMA)-b-PVP]4) were synthesized after polymerization reaction of double bones between RhB-HEMA and other reactants. Finally, the structure of product was measured by FT-IR spectra and fluorospectro photometer (FLUORO). The critical micelle concentration of RhB-labeled and unlabeled amphipathic block polymers were 4.96×10-3, 5.09×10-3 mg·L-1, respectively, indicating no change of their micellization behavior. In vivo tissue distribution and whole-body fluorescent imaging were studied by vinpocetine (VP)-loaded polymeric micelles which were prepared through a solvent evaporation method. Compared to the result of in vivo tissue distribution and whole-body fluorescence imaging, a similar bio-distribution behavior of VP-loaded polymeric micelles was found. Those proved the successful fluorescence modification with a labeling yield of 4.13%. With in vivo fluorescence imaging technology, we established a fluorescence method for modification of amphipathic block polymers.

Relation between drug release and the drug status within curcumin-loaded microsphere / 药学学报

To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21) %, (11.71 ± 0.39) %, (15.41 ± 0.40) %, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64) % and (5.66 ± 0.07) % curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.

Relation between drug release and the drug status within curcumin-loaded microsphere / 药学学报

To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.

Tetrahydrobiopterin Protects against Radiation-induced Growth Inhibition in H9c2 Cardiomyocytes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tetrahydrobiopterin (BH4) is an essential cofactor of nitric oxide synthases (NOSs) for the synthesis of nitric oxide (NO). BH4 therapy can reverse the disease-related redox disequilibrium observed with BH4 deficiency. However, whether BH4 exerts a protective effect against radiation-induced damage to cardiomyocytes remains unknown.</p><p><b>METHODS</b>Clonogenic assays were performed to determine the effects of X-ray on H9c2 cells with or without BH4 treatment. The contents of lactate dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA) in H9c2 cells were measured to investigate oxidative stress levels. The cell cycle undergoing radiation with or without BH4 treatment was detected using flow cytometry. The expression levels of proteins in the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT)/P53 signaling pathway, inducible NOS (iNOS), and endothelial NOS (eNOS) were examined using Western blotting.</p><p><b>RESULTS</b>X-ray radiation significantly inhibited the growth of H9c2 cells in a dose-dependent manner, whereas BH4 treatment significantly reduced the X-ray radiation-induced growth inhibition (control group vs. X-ray groups, respectively, P< 0.01). X-ray radiation induced LDH release, apoptosis, and G0/G1 peak accumulation, significantly increasing the level of MDA and the production of NO, and decreased the level of SOD (control group vs. X-ray groups, respectively, P < 0.05 or P < 0.01). By contrast, BH4 treatment can significantly reverse these processes (BH4 treatment groups vs. X-ray groups, P < 0.05 or P < 0.01). BH4 reversed the X-ray radiation-induced expression alterations of apoptosis-related molecules, including B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein, and caspase-3, and molecules of the PI3K/Akt/P53 signaling pathway. BH4 enhanced the production of NO in 2 Gy and 4 Gy radiated groups by upregulating eNOS protein expression and downregulating iNOS protein expression.</p><p><b>CONCLUSIONS</b>BH4 treatment can protect against X-ray-induced cardiomyocyte injury, possibly by recoupling eNOS rather than iNOS. BH4 treatment also decreased oxidative stress in radiated H9c2 cells.</p>

Sox2 function as a negative regulator to control HAMP expression

BACKGROUND: Hepcidin, encoding by HAMP gene, is the pivotal regulator of iron metabolism, controlling the systemic absorption and transportation of irons from intracellular stores. Abnormal levels of HAMP expression alter plasma iron parameters and lead to iron metabolism disorders. Therefore,itis animportant goal to understand the mechanisms controlling HAMP gene expression. RESULTS: Overexpression of Sox2 decrease basal expression of HAMP or induced by IL-6 or BMP-2, whereas, knockdown of Sox2 can increase HAMP expression, furthermore, two potential Sox2-binding sites were identified within the human HAMP promoter. Indeed, luciferase experiments demonstrated that deletion of any Sox2-binding site impaired the negative regulation of Sox2 on HAMP promoter transcriptional activity in basal conditions. ChIP experiments showed that Sox2 could directly bind to these sites. Finally, we verified the role of Sox2 to negatively regulate HAMP expression in human primary hepatocytes. CONCLUSION: We found that Sox2 as a novel factor to bind with HAMP promoter to negatively regulate HAMP expression, which may be further implicated as a therapeutic option for the amelioration of HAMP-overexpression-related diseases, including iron deficiency anemia.

Activity and expression of aldehyde dehydrogenases in the ocular tissue of diabetic rats / 中华实验眼科杂志

Background Diabetic complication is associated with lipid peroxidation.Aldehyde dehydrogenases (ALDH) catalyze the irreversible oxidation of a variety of biological aldehydes,including lipid-derived aldehydes (LDAs),and thus protect organs and tissues from toxic LDAs.Understanding the activity of ALDH in different ocular tissues in diabetic subjects is very important for prevention and treatment of diabetic ocular complications.Objective This research aimed to investigate the activity and expression of ALDH in different ocular tissues in diabetic rats and to explore the mechanism of ALDH in diabetes-induced eye disease.Methods Twenty-eight healthy SPF male Sprague-Dawley(SD) rats weighted 170-180 g were randomly divided into the normal control group and diabetic group.The diabetic animal model was established by intraperitonial injection of 4％ streptozotocin at 65 mg/kg.Isometric citric acid buffer was injected in the rats of the normal control group.The rats were sacrificed in each group 2 and 4 months after the establishment of the diabetic models,and eyeballs were obtained for the preparation of corneal,lens and retinal homogenates.ALDH activity was detected using a multifunctional microplate reader SpectraMax M5,and ALDH content was measured by ELISA at the wavelength of 450 nm with the SpectraMax M5 ELISA reader.Results The blood glucose level in diabetic rats was significantly elevated at various time points compared with the normal control group(P=0.000),and body weights were evidently lower in the diabetic group than in the normal group (P =0.000).The activities of ALDH (A340) in corneal,lens and retinal tissues in the diabetic group were increased in comparison with the normal control group (F =396.601,P=0.000),and showed an enhancement with the lapsing of time (F =53.139,P =0.000).In addition,the highest level of ALDH was found in the cornea and the lowest level in the lens(F =6973.000,P=0.000).The expression level of ALDH in the corneal,lens and retinal homogenates was significantly higher in the diabetic group compared with the normal control group (F=312.985,P =0.000) and showed a considerable increase over the course (F =19.203,P=0.000).The highest expression level was seen in the cornea and the lowest was in the lens,with a significant difference among these three kinds of tissues (F =3243.000,P =0.000).Conclusions ALDH can protect ocular tissue from the damage of lipid peroxidation.Thess results suggest that ALDH plays a role in preventing diabetes-related ocular complications.