Structure-activity relationship of Lycium barbarum polysaccharides / 中国中药杂志

As a traditional Chinese herb and functional food, the fruits of Lycium barbarum has been widely used for thousands of years in China. L. barbarum polysaccharides(LBPs) are predominant active components, which have immunomodulatory, antioxidant, hypoglycemic, neuroprotective, anti-tumor, and prebiotic activities. The molecular weight, monosaccharide composition, glycosidic bond, branching degree, protein content, chemical modification, and spatial structure of LBPs are closely related to their biological activity. Based on the previous studies of this research team, this paper systematically combed and integrated the research progress of structure, function, and structure-activity relationship of LBPs. At the same time, some problems restricting the clarification of the structure-activity relationship of LBPs were considered and prospected, hoping to provide references for the high value utilization of LBPs and in-depth exploration of their health value.

Effects of porcine acellular dermal matrix combined with human epidermal stem cells on wound healing of full-thickness skin defect in nude mice / 中华烧伤杂志

Objective: To explore the effects of porcine acellular dermal matrix (ADM) combined with human epidermal stem cells (ESCs) on wound healing of full-thickness skin defect in nude mice. Methods: The morphology of porcine ADM was analyzed by photograph of digital camera, the cell residues in porcine ADM were observed by hematoxylin-eosin (HE) staining, the surface structure of porcine ADM was observed by scanning electron microscope, the secondary structure of porcine ADM was analyzed by infrared spectrometer, the porcine ADM particle size was analyzed by dynamic light scattering particle size analyzer, and the porcine ADM potential was analyzed by nano-particle size potentiometer. The morphology of porcine ADM was observed by inverted fluorescence microscope when it was placed in culture medium for 30 min, 1 d, and 5 d (n=2). The porcine ADM was divided into 5 min group, 10 min group, 20 min group, 30 min group, 60 min group, and 120 min group according to the random number table (the same grouping method below) in static state at normal temperature for the corresponding time to calculate the water absorption by weighing method (n=3). Swiss white mouse embryonic fibroblasts (Fbs) were divided into blank control group (culture medium only), and 50.0 g/L ADM extract group, 37.5 g/L ADM extract group, 25.0 g/L ADM extract group, 12.5 g/L ADM extract group, and 6.5 g/L ADM extract group which were added with the corresponding final concentrations of ADM extract respectively. At post culture hour (PCH) 24, 48, and 72, the cell survival rate was detected by cell counting kit 8 and the cytotoxicity was graded (n=5). The erythrocytes of a 6-week-old male Sprague-Dawley male rat were divided into normal saline group, ultra-pure water group, and 5 mg/mL ADM extract group, 10 mg/mL ADM extract group, and 15 mg/mL ADM extract group which were treated with the corresponding final concentrations of porcine ADM extract respectively. After reaction for 3 h, the absorbance value of hemoglobin was detected by microplate reader to represent the blood compatibility of porcine ADM (n=3). ESCs were isolated and cultured from the discarded prepuce of a 6-year-old healthy boy who was treated in the Department of Urology of the First Affiliated Hospital of Army Medical University (the Third Military Medical University) in July 2020, and then identified by flow cytometry. The porcine ADM particles of composite ESC (hereinafter referred to as ESC/ADM) were constructed by mixed culture. After 3 days of culture, the composite effect of ESC/ADM was observed by HE staining and laser scanning confocal microscope. Thirty-six 7-8-week-old male non-thymic nude mice were divided into phosphate buffer solution (PBS) alone group, ADM alone group, ESC alone group, and ESC/ADM group, with 9 mice in each group, and the wound model of full-thickness skin defect was established. Immediately after injury, the wounds were treated with the corresponding reagents at one time. On post injury day (PID) 1, 7, 11, and 15, the wound healing was observed and the wound healing rate was counted (n=3). On PID 7, the epithelialization of wounds was observed by HE staining and the length of un-epithelialized wound was measured (with this and the following sample numbers of 4). On PID 11, the dermal area and collagen deposition of wounds were observed by Masson staining and the dermal area of wound section was calculated, the number of cells expressing CD49f, a specific marker of ESC, was calculated with immunofluorescence staining, the mRNA expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in ESC after wound transplantation was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with independent sample t test, one-way analysis of variance, analysis of variance for repeated measurement, and least significant difference t test. Results: The porcine ADM was white particles and composed of reticular structure, with no cells inside, disordered structure, and rough surface. The absorption peak of porcine ADM appeared at the wave numbers of 1 659, 1 549, and 1 239 cm-1, respectively. The main particle size distribution of porcine ADM in solution was 500 to 700 nm, with negative charge on the surface. The morphology of porcine ADM in static state at 30 min and on 1 and 5 d was relatively stable. The water absorption of porcine ADM remained relatively high level in static state from 30 min to 120 min. The cytotoxicity of mouse embryonic Fbs in 6.5 g/L ADM extract group, 12.5 g/L ADM extract group, and 25.0 g/L ADM extract group was grade 1 at PCH 24, and the cytotoxicity of the other groups was 0 grade at each time point. After reaction for 3 h, the absorbance value of hemoglobin of erythrocytes in ultra-pure water group was significantly higher than the values in normal saline group and 15 mg/mL ADM extract group (with t values of 8.14 and 7.96, respectively, P<0.01). After 3 days of culture, the cells of the fourth passage showed pebble-like morphology, with low expression of CD71 and high expression of CD49f, which were identified as ESCs. There was ESC attachment and growth on porcine ADM particles. On PID 1, the wound sizes of nude mice were almost the same in PBS alone group, ADM alone group, ESC alone group, and ESC/ADM group. On PID 7, 11, and 15, the wound contraction of nude mice in each group was observed, especially in ADM alone group, ESC alone group, and ESC/ADM group. On PID 7, the wound healing rates of nude mice in ESC alone group and ESC/ADM group were significantly higher than the rate in PBS alone group (with t values of 2.83 and 4.72 respectively, P<0.05 or P<0.01). On PID 11, the wound healing rate of nude mice in ESC/ADM group was significantly higher than that in PBS alone group (t=4.86, P<0.01). On PID 15, the wound healing rates of nude mice in ADM alone group, ESC alone group, and ESC/ADM group were significantly higher than the rate in PBS alone group (with t values of 2.71, 2.90, and 3.23 respectively, P<0.05). On PID 7, the length of un-epithelialized wound of nude mice in ADM alone group, ESC alone group, and ESC/ADM group was (816±85), (635±66), and (163±32) μm, respectively, which were significantly shorter than (1 199±43) μm in PBS alone group (with t values of 5.69, 10.19, and 27.54 respectively, P<0.01). On PID 11, the dermal areas of wound section of nude mice in ADM alone group, ESC alone group, and ESC/ADM group were significantly larger than the area in PBS alone group (with t values of 27.14, 5.29, and 15.90 respectively, P<0.01); the collagen production of nude mice in ADM alone group and ESC/ADM group was more obvious than that in PBS alone group, and the collagen production of nude mice in ESC alone group and PBS alone group was similar. On PID 11, in the wounds of nude mice in ESC alone group and ESC/ADM group, the cells with positive expression of CD49f were respectively 135±7 and 185±15, and the mRNA expressions of GAPDH were positive; while there were no expressions of CD49f nor mRNA of GAPDH in the wounds of nude mice in PBS alone group and ADM alone group. Conclusions: ESC/ADM particles can promote the wound healing of full-thickness skin defects in nude mice, which may be related to the improved survival rate of ESCs after transplantation and the promotion of dermal structure rearrangement and angiogenesis by ADM.

Identification of key genes in nonarteritic anteriorischemic optic neuropathy through weighted gene co-expression network analysis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: We sought to identify key genes related to nonarteritic anterior ischemic optic neuropathy(NAION)and provide bioinformatics support for elucidating the pathogenesis of NAION.METHODS: Based on rat GSE43671 dataset, which was acquired from GEO, we identified modular genes with highly correlated clinical phenotype by WGCNA package in the R language. Then Gene Ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)analysis were performed with ClusterProfiler package. In addition, Cytoscape was used to screen potential key genes and establish miRNA-key genes network.RESULTS: There were 22 modules identified from the GSE43671 dataset by the WGCNA method, among which the blue module has the highest correlation coefficient. GO enrichment analysis suggested that the genes in the module mainly manifest in the epithelial tube morphogenesis and other biological processes, receptor complex and other cell components, and structural constituent of eye lens and other molecular functions. KEGG suggested that the genes in the module mainly relate to signaling pathways including neuroactive ligand-receptor interaction, human papillomavirus, MAPK and PI3K/Akt. There were 10 key genes screened by PPI network and Cytoscape including Psmb9, Psma7, Map3k14, Psme1, Nfkb1, Rela, Psma5, Relb, Psmb4 and Nfkb2, and 6 miRNA were predicted as miR-383-5p, miR-9a-5p, miR-155-5p, miR-223-3p, miR-495 and miR-325-3p.CONCLUSION: Using the WGCNA method to screen out the relevant pathways, key genes, and microRNA for NAION, it provides a theoretical basis for exploring pathogenesis and treatment methods of NAION, however, more animal and cell experiments are needed to further validate.

Comparison of Murraya exotica and Murraya paniculata by fingerprint analysis coupled with chemometrics and network pharmacology methods / 中国天然药物

There are two source plants for the traditional Chinese medicine Murrayae Folium et Cacumen (MFC) in Chinese Pharmacopoeia, i.e. Murraya exotica L. and M. paniculata (L.) Jack. Herein, a chemical comparison of M. exotica and M. paniculata by high performance liquid chromatography (HPLC) fingerprint analysis coupled with chemometrics and network pharmacology was performed. The main peaks in the fingerprints were identified by liquid chromatography coupled with ion trap/time-of-flight mass spectrometry (LC-IT-TOF-MS) and authenticated by references. The chemometrics results showed that the HPLC fingerprints of these two species were clearly divided into two categories using hierarchical cluster analysis (HCA) and principal component analysis (PCA), and a total of 13 significantly differentiated markers were screened out by orthogonal partial least squares-discriminant analysis (OPLS-DA). However, the following network pharmacology analysis showed that these discriminated markers were found to act via many common targets and metabolic pathways, indicating the possibly similar pharmacological effects and mechanisms for M. exotica and M. paniculata. The above results provide valuable evidence for the equivalent use of these two plants in clinical settings. Moreover, the chromatographic fingerprint analysis coupled with chemometrics and network pharmacology supplies an efficient approach for the comparative analysis of multi-source TCMs like MFC.

Historical evolution of Salviae Miltiorrhizae Radix et Rhizoma processing methods / 中国中药杂志

China has a long history of Salviae Miltiorrhizae Radix et Rhizoma processing with multiple methods available. The pre-sent study collated and summarized the Salviae Miltiorrhizae Radix et Rhizoma processing methods recorded in 23 related herbal medicine books, all editions of Chinese Pharmacopoeia, the 1988 edition of National Regulations for Processing of Chinese Medicine, and 20 current local processing specifications and standards. The results demonstrated various processing methods of Salviae Miltiorrhizae Radix et Rhizoma, such as removing residual part of stem, plantlet, or soil, smashing, filing, cutting, decocting, washing with wine, soaking in wine, and stir-frying with wine or blood from pig heart, while raw and wine-processed products are mainly used in modern times. Due to the lack of unified standards, the phenomena of multiple methods adopted in one place and different methods in different places have led to uneven quality of Salviae Miltiorrhizae Radix et Rhizoma pieces, even affecting the safety and effectiveness of its clinical medication. This study is expected to provide a reference for the development of Salviae Miltiorrhizae Radix et Rhizoma processing and its rational medication.

Effects of Renshenjian Decoction on blood and urine metabonomics in type 2 diabetes rats based on ~1H-NMR / 中国中药杂志

Proton nuclear magnetic resonance(~1H-NMR) is used to investigate the effect of Renshenjian Decoction on serum and urine metabolism of type 2 diabetic rats with insulin resistance induced by high-sugar and high-fat diet combined with low-dose streptozotocin(STZ). After the successful establishment of the insulin resistance model of type 2 diabetes, administration for 35 days, the serum and urine of rats were taken. Once the ~1H-NMR data have been collected and processed, PCA and OPLS-DA were used to analyze them. The results show that: compared with the blank group, the contents of methionine, taurine, α-glucose and β-glucose in the serum of the model group increased significantly(P<0.001), while the contents of 3-hydroxybutyric acid, lactic acid and unsaturated fatty acids decreased significantly(P<0.01). In the model group, the contents of trimethylamine oxide, glycine, α-glucose, β-glucose, taurine and phosphocholine in urine increased significantly(P<0.05), while the contents of creatine, lactic acid, acetic acid and citric acid decreased significantly(P<0.05). Compared with the model group, the contents of 3-hydroxybutyric acid and unsaturated fatty acids in serum of rats in the treatment group increased significantly(P<0.05), while the contents of taurine, α-glucose and β-glucose decreased significantly(P<0.01). In the treatment group, the contents of lactic acid, taurine and creatine in urine increased significantly(P<0.05), while the contents of trimethylamine oxide, glycine, α-glucose, β-glucose and phosphocholine decreased significantly(P<0.01). The results show that Renshenjian Decoction can regulate metabolic disorder and promote the metabolic phenotype to return to the normal range. It displayed therapeutic effect on type 2 diabetic rats with insulin resistance and provided a certain scientific basis for the biological basic research of Renshenjian Decoction by improving insulin resistance in diabetes mellitus.

Effects of RNA Targeting mPGES-1 on Proliferation and Apoptosis of K562/A Cells / 中山大学学报(医学科学版)

@#【Objective】 To explore the effects and the possible mechanism of RNA targeting membrane-bound prostaglandin E2 synthase l（mPGES- 1）on proliferation，apoptosis and drug resistance of leukemia cell line K562/A.【Methods】RNA interference was used to inhibit the expression of mPGES-1 of K562/A cells. Four groups were set up as follows：untreated group（K562/A），negative control group after interference（K562/A-NC），group after interference（K562/ A-KD），and group after interference with exogenous PGE2（K562/A-KD+PGE2）.Cell viability was assessed by CCK-8 assay. Cell apoptosis was analyzed by flow cytometry. Concentration of PGE2 was detected by ELISA. Proteins expression was detected by western blot.【Results】The expression of mPGES- 1 in K562/A cells was significantly down- regulated and the synthesis of PGE2 decreased（P < 0.000 1）after RNA interference. After RNA interference，the proliferation of K562/A cells was inhibited and apoptosis increased，and the sensitivity to chemotherapy drugs was enhanced（P < 0.05）. Meanwhile，the expression of β-catenin and MDR1 was decreased（P < 0.01）. Exogenous PGE2 could reverse the effect of RNA interference on proliferation ，apoptosis and drug sensitivity in K562/A cells（P < 0.05），and up-regulate the expression of β-catenin and MDR1（P < 0.01）. XAV939，an inhibitor of β-catenin，could down-regulate the expression of β- catenin and MDR1 in an dose- dependent pattern in K562/A cells（P < 0.05）.【Conclusions】RNA interference of mPGES- 1 could inhibit proliferation，induce apoptosis and reverse drug resistance in K562/A cells. The mechanism was related to reducing the synthesis of PGE2 and thus down- regulating the expression of β- catenin and MDR1. Wnt/β- catenin signal pathway may participate in the regulation of MDR1 by mPGES-1/PGE2.

Research progress of receptor protein FUNDC1 in mitophagy / 医学研究生学报

Mitochondria, which play an important role in cell metabolism, stress response and cell death, are the key organelles that regulate the energy balance of cells. Under the influence of internal and external environment, damaged or senescent mitochondria pose a serious threat to cell survival. Mitophagy refers to the selective elimination of dysfunctional mitochondria to maintain the homeostasis of the intracellular environment. FUN14 domain containing 1 (FUNDC1) is a newly discovered mitophagy receptor protein, which plays an important regulatory role in mediating mitophagy. This paper mainly reviews the recent research progress of FUNDC1 regulation mechanism and its pathophysiological significance in mitophagy.

Relationship between depressive symptoms andblood biochemical indices in elderly inpatients / 中国心理卫生杂志

Objective:To investigate the relationship between the depressive symptoms and blood biochemical indexes in elderly inpatients.Methods:Totally 201 inpatients without depression were enrolled in this study.Blood tests and the Geriatric Depression Scale-5 (GDS-15) were performed on all of the patients.The patients were divided into non-depressive symptoms group and depressive symptoms group according to GDS-15 score.Differences between two groups were compared, and multiple linear regression analysis was used to screen for risk factors by SPSS 22.0.Results:Serum total protein (TP) [ (60.7±4.3) vs. (63.5±5.6) g/L] and hemoglobin [ (121.6±20.1) vs. (129.7±19.4) g/L] were lower in the depressive symptoms group than in the non-depressive symptoms group (Ps＜0.05).There were no differences between the two groups in age, BMI, fasting blood glucose, total cholesterol, triglyceride, high density lipoprotein cholesterol and low density lipoprotein cholesterol (Ps＞0.05).Multiple linear regression analysis showed that TP was the predictive variable for GDS-15 score (β=-0.4, P＜0.05).Conclusion:Serum total protein and hemoglobin levels in patients with depressive symptoms may be significant lower than those in patients without depressive symptom.There is association between serum total protein and depressive symptoms.

Relationship between the CT features of colorectal cancer metastases calcification and tumor response to chemotherapy / 北京大学学报(医学版)

OBJECTIVE@#To investigate the relationship between CT features of metastatic calcification and the response to chemotherapy in colorectal cancer metastases.@*METHODS@#A total of 27 patients with 30 sites of calcified metastases who underwent chemotherapy combined with targeted therapy (cetuximab) between January 2011 and December 2016 comprised this retrospective study population. Two radiologists independently evaluated the occurrence of tumor calcification before and after treatment, and evaluated the tumor response after therapy. According to the response evaluation criteria in solid tumors (version 1.1), the best curative effect evaluation of the patients was recorded. The patients were divided into groups as below: (1) Patients who showed complete response (CR) and partial response (PR) were assigned to the response group, and the stable disease (SD) and progressive disease (PD) were assigned to the non-response group. (2) Patients showed CR or PR, or patients showed SD with longer progress free survival (PFS) were assigned to the benefit group, and the remaining patients were assigned to the no benefit group. The difference of different imaging calcification features (morphology, maximum density, and density-time slope) were analyzed.@*RESULTS@#The most common site of metastases calcification was liver (63.3%), followed by lymph nodes (26.7%). There were 12 cases in the response group, 15 cases in the non-response group; and 13 cases in the benefit group, 14 cases in the no benefit group. The density time growth slope was higher in the response group when compared with the non-response group (P=0.025). The proportion of thhe patients with increased number of calcified foci in the benefit group (61.5%) was higher than that in the no benefit group (14.3%), P=0.018. There was no significant difference in the maximum density between the groups. The calcification of liver metastases were all amorphous calcification, with central calcification (36.8%), eccentric calcification (36.8%), garland calcification (15.8%) and diffuse calcification (10.6%). The lymph node metastases could be diffuse (75.0%), and curve or eggshell calcification (25.0%). There was no statistical difference between the groups.@*CONCLUSION@#In patients with advanced colorectal cancer metastases treated with cetuximab combined chemotherapy, rapid growth of calcification density and increased calcification number may be valuable imaging features of therapeutic efficacy. The maximal calcification density and morphology of calcification are not related to the therapeutic efficacy.

Effect of NEU3 on biological behaviors of human prostate cancer DU145 cells / 中国病理生理杂志

AIM:To explore the effects of neuraminidase 3 (NEU3) on the viability, invasion and apoptosis of human prostate cancer DU145 cells and the molecular mechanism.METHODS:The human prostate cancer DU145 cells were divided into blank control group and treatment group.The cells in treatment group were treated with either neuraminidase inhibitor DANA, or NEU3 small interfering RNA (siRNA) to knock down the expression of NEU3.The cell viability was measured by CCK-8 assay.The cell invasion ability was detected by Transwell assay.The effects of the treatments on the mRNA level of Bcl-2 were detected by q PCR.The effects of the treatments on the protein levels of matrix metalloproteinase 2 (MMP2) and apoptotic inhibitory protein Bcl-2 were determined by Western blot.Apoptosis of the cells was analyzed by flow cytometry.RESULTS:The protein level of NEU3 and the apoptotic rate in DANA group were not significantly different from those in blank control group.The viability of DANA-treated DU145 cells was increased, and the invasion ability, MMP2 protein level, and Bcl-2 mRNA and protein levels were all decreased in these cells, compared with blank control group.On the other hand, the levels of NEU3 protein and Bcl-2 mRNA and protein in NEU3 siRNA group were significantly decreased compared with blank control group, while the viability and apoptotic rate of the cells with NEU3 siRNA transfection were increased (P<0.05).However, the protein expression of MMP2 and the invasion ability of the cells were not significantly changed after NEU3 siRNA treatment.CONCLUSION:The inhibition of NEU3 in enzyme activity and expression decreases the viability, and enhances the apoptosis of human prostate cancer DU145 cells.However, it has no obvious effect on the invasion ability of DU145 cells.

Cryo-EM structure of an early precursor of large ribosomal subunit reveals a half-assembled intermediate

Assembly of eukaryotic ribosome is a complicated and dynamic process that involves a series of intermediates. It is unknown how the highly intertwined structure of 60S large ribosomal subunits is established. Here, we report the structure of an early nucleolar pre-60S ribosome determined by cryo-electron microscopy at 3.7 Å resolution, revealing a half-assembled subunit. Domains I, II and VI of 25S/5.8S rRNA pack tightly into a native-like substructure, but domains III, IV and V are not assembled. The structure contains 12 assembly factors and 19 ribosomal proteins, many of which are required for early processing of large subunit rRNA. The Brx1-Ebp2 complex would interfere with the assembly of domains IV and V. Rpf1, Mak16, Nsa1 and Rrp1 form a cluster that consolidates the joining of domains I and II. Our structure reveals a key intermediate on the path to establishing the global architecture of 60S subunits.

Prediction of the therapeutic response after target-combined chemotherapy treatment for patients with liver metastasis from colorectal cancer using computed tomography texture analysis / 生物医学工程学杂志

This study aims to investigate the value of pre-treatment computed tomography (CT) texture analysis in predicting therapeutic response of liver metastasis from colorectal cancer after combined targeting chemotherapy. A total of 82 patients with colorectal cancer liver metastases who underwent chemotherapy combined with targeted therapy (cetuximab) between March 2011 and October 2017 comprised this retrospective study population. According to the RECIST1.1, the best curative effect evaluation of patients was recorded. Complete response (CR) and partial response (PR) were assigned to the response group, and the stable disease (SD) and progressive disease (PD) were assigned to the non-response group. The CT texture analysis was based on the Omini-Kinetics software, and the three-dimensional (3D) texture analysis was performed on the marked lesion on portal phase. The differences of texture parameters between the response group and the non-response group were compared. The receiver operating characteristic (ROC) curves were depicted on the parameters which with statistically difference, to characterize value in predicting the response to target-combined chemotherapy. The differences of Entropy, Energy, Variance, std. Deviation, Quantile95 and sumEntropy between the two groups in pre-treatment lesions were significant ( 0.867, good diagnostic efficiency could be obtained, with sensitivity of 60.5% and specificity of 79.5%, respectively. In conclusion, texture parameters derived from baseline CT images of colorectal cancer liver metastasis have the potential value acting as imaging biomarkers in predicting tumor response to combined target chemotherapy.

Effect of salvianolic acid B on vasodilatory function, NF-κB activation and inflammatory cytokine expression in diabetic rats / 中国病理生理杂志

AIM:To investigate the ameliorative effect of salvianolic acid B on vasodilatory function in diabetic rats and the possible mechanisms.METHODS:SD rats(n=40)were fed on high-sugar and high-fat diet for 4 weeks, followed by a single intraperitoneal injection of streptozotocin(40 mg/kg).The rats with random blood glucose level over 16.7 mmol/L were considered diabetic and randomly allocated to 3 groups, namely model group, low dose(80 mg· kg-1· d-1)of salvianolic acid B group and high dose(160 mg· kg-1· d-1)of salvianolic acid B group.The rats in salvianolic acid B groups were intragastrically administered with corresponding doses of salvianolic acid B for 6 weeks. Vasodilatory function was measured as endothelium-dependent and-independent vasodilation of the aortic rings.The primary histopathological changes of aorta were observed by HE staining.Serum levels of interleukin-6(IL-6),tumor necrosis fac-tor-α(TNF-α)and C-reactive protein(CRP)were measured by ELISA.The levels of total antioxidant capacity,malondi-aldehyde(MDA)and nitric oxide(NO)in aortic tissues were evaluated by colorimetric assays.The protein levels of inter-cellular adhesion molecule-1(ICAM-1)and monocyte chemotactic protein-1(MCP-1), and the activation of nuclear fac-tor-κB(NF-κB)were determined by Western blot.RESULTS: Treatment with salvianolic acid B evidently ameliorated endothelium-dependent diastolic function and pathological changes of aorta in diabetic rats(P<0.05 or P<0.01).Sup-plementation with salvianolic acid B resulted in significant increases in NO content and total antioxidant capacity in aortic tissues,accompanied by marked decreases in the level of MDA in aorta tissues and the serum levels of IL -6, TNF-αand CRP(P<0.05 or P<0.01).Salvianolic acid B markedly down-regulated NF-κB p65 nuclear translocation and protein expression of ICAM-1 and MCP-1 in aorta tissues(P<0.05 or P<0.01).CONCLUSION:Salvianolic acid B effectively ameliorates endothelium-dependent diastolic function of aorta in diabetic rats, which might be attributed to suppression of NF-κB activation and subsequent expression of inflammatory cytokines.The beneficial effect of salvianolic acid B on vascu-lar endothelium might be derived from its antioxidant capacity.

Expression of matrix metalloproteinase-9, tissue inhibitor of metallopro-teinase-1 and collagen type Ⅳ in lung of rats with multiple organ dysfunc-tion syndrome / 中国病理生理杂志

AIM:To determine the expression of matrix metalloproteinase-9(MMP-9),tissue inhibitor of me-talloproteinase-1(TIMP-1)and collagen type Ⅳ(Ⅳ-C)in the lung of rats with multiple organ dysfunction syndrome(MODS)and to investigate the mechanism of lung injury in MODS.METHODS:Adult male Sprague-Dawley(SD)rats(n=40)were randomly divided into sham control group and cecal ligation and puncture(CLP)model group.The rats in CLP group were divided into 4 subgroups as different intervals(6 h,12 h,24 h and 48 h),and there were 8 rats in each group.The rat model of MODS was established by CLP.All rats were sacrificed at various intervals.The functions of the liver,kidney and lung were determined by blood biochemical and blood gas analysis.The morphological changes of the lung tissues were observed with HE staining.The serum levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,MMP-9 and TIMP-1 were measured by ELISA.The expression of MMP-9 and TIMP-1 in the lung tissues was detected by RT-PCR and immunohistochemistry ,and the expression of Ⅳ-C in the lung tissues was detected by immunofluorescence and Western blot.RESULTS:Compared with sham control group ,the functions of the liver ,kidney and lung were damaged at different degrees in model groups.No histopathological change in the lung tissues of sham control group was found ,and the lung injury was serious in model groups.Compared with sham control group ,the serum levels of TNF-α,IL-1β,MMP-9 and TIMP-1 in model groups increased significantly(P<0.05)and peaked at the interval of 12 ～24 h after modeling(P<0.01).The expression of MMP-9 and TIMP-1 in the lung tissues of model groups increased ,and peaked at 12 and 24 h,respectively(P<0.01).The protein level of Ⅳ-C in MODS 6 h group was not changed as compared with control group,while that at the interval of 12～48 h after modeling was significantly decreased and dropped to the lowest at 24 h(P<0.01).CONCLUSION:MMP-9 and TIMP-1 play important roles in lung injury of MODS rats by regulating the syn-thesis and decomposition of IV-C which is the main component of extracellular matrix.

Diagnostic Value of Cerebrospinal Fluid T-SPOT.TB for Tuberculousis Meningitis in China / 生物医学与环境科学（英文）

The aim of this study was to evaluate the diagnostic value of the cerebrospinal fluid (CSF) T-SPOT.TB test for the diagnosis of TB meningitis (TBM). A retrospective analysis of 96 patients with manifested meningitis was conducted; T-SPOT.TB test was performed for diagnosing TBM to determine the diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). A receiver operating characteristic (ROC) curve was also drawn to assess the diagnostic accuracy. The sensitivity, specificity, PPV, and NPV of CSF T-SPOT.TB test were 97.8%, 78.0%, 80.3%, and 97.5%, respectively, for 52 patients (54.2%) of the 96 enrolled patients. The area under the curve (AUC) was 0.910, and the sensitivities of CSF T-SPOT.TB for patients with stages I, II, and III of TBM were 96.7%, 97.2%, and 98.9%, respectively. CSF T-SPOT.TB test is a rapid and accurate diagnostic method with higher sensitivity and specificity for diagnosing TBM.

Structural characterization of coatomer in its cytosolic state

Studies on coat protein I (COPI) have contributed to a basic understanding of how coat proteins generate vesicles to initiate intracellular transport. The core component of the COPI complex is coatomer, which is a multimeric complex that needs to be recruited from the cytosol to membrane in order to function in membrane bending and cargo sorting. Previous structural studies on the clathrin adaptors have found that membrane recruitment induces a large conformational change in promoting their role in cargo sorting. Here, pursuing negative-stain electron microscopy coupled with single-particle analyses, and also performing CXMS (chemical cross-linking coupled with mass spectrometry) for validation, we have reconstructed the structure of coatomer in its soluble form. When compared to the previously elucidated structure of coatomer in its membrane-bound form we do not observe a large conformational change. Thus, the result uncovers a key difference between how COPI versus clathrin coats are regulated by membrane recruitment.

4.4 Å Resolution Cryo-EM structure of human mTOR Complex 1

Mechanistic target of rapamycin (mTOR) complex 1 (mTORC1) integrates signals from growth factors, cellular energy levels, stress and amino acids to control cell growth and proliferation through regulating translation, autophagy and metabolism. Here we determined the cryo-electron microscopy structure of human mTORC1 at 4.4 Å resolution. The mTORC1 comprises a dimer of heterotrimer (mTOR-Raptor-mLST8) mediated by the mTOR protein. The complex adopts a hollow rhomboid shape with 2-fold symmetry. Notably, mTORC1 shows intrinsic conformational dynamics. Within the complex, the conserved N-terminal caspase-like domain of Raptor faces toward the catalytic cavity of the kinase domain of mTOR. Raptor shows no caspase activity and therefore may bind to TOS motif for substrate recognition. Structural analysis indicates that FKBP12-Rapamycin may generate steric hindrance for substrate entry to the catalytic cavity of mTORC1. The structure provides a basis to understand the assembly of mTORC1 and a framework to characterize the regulatory mechanism of mTORC1 pathway.

Structural dynamics of the yeast Shwachman-Diamond syndrome protein (Sdo1) on the ribosome and its implication in the 60S subunit maturation

The human Shwachman-Diamond syndrome (SDS) is an autosomal recessive disease caused by mutations in a highly conserved ribosome assembly factor SBDS. The functional role of SBDS is to cooperate with another assembly factor, elongation factor 1-like (Efl1), to promote the release of eukaryotic initiation factor 6 (eIF6) from the late-stage cytoplasmic 60S precursors. In the present work, we characterized, both biochemically and structurally, the interaction between the 60S subunit and SBDS protein (Sdo1p) from yeast. Our data show that Sdo1p interacts tightly with the mature 60S subunit in vitro through its domain I and II, and is capable of bridging two 60S subunits to form a stable 2:2 dimer. Structural analysis indicates that Sdo1p bind to the ribosomal P-site, in the proximity of uL16 and uL5, and with direct contact to H69 and H38. The dynamic nature of Sdo1p on the 60S subunit, together with its strategic binding position, suggests a surveillance role of Sdo1p in monitoring the conformational maturation of the ribosomal P-site. Altogether, our data support a conformational signal-relay cascade during late-stage 60S maturation, involving uL16, Sdo1p, and Efl1p, which interrogates the functional P-site to control the departure of the anti-association factor eIF6.

Intervention effectiveness of intensive training in medical interns’occupa-tional exposure protection / 中国感染控制杂志

Objective To study the intervention effectiveness of intensive training in medical interns’occupational exposure protection.Methods Medical interns in seven colleges and universities in Wuhan between 2011 and 2015 were chosen and divided into control group (675 medical interns in 2011 - 2013,comprehensive training about healthcare-associated infection control was conducted for once before internship )and intervention group (682 medical interns in 2014 - 2015,hospital-department-tutor three steps of intensive training was conducted),the occurrence of occupational exposure and exposure knowledge awareness between two groups were compared. Results The awareness rate of knowledge about occupational exposure protection and the implementation rate of protective measures in intervention group were both higher than control group (all P <0.05).Incidence of occupa-tional exposure in intervention group was lower than control group (21 .70%[n=148]vs 65.33%[n=441 ],χ2 =262.91 ,P <0.01);percentage of interns who conducted active serological virus detection and intensified vaccination of hepatitis B virus were both higher than control group (14.66%[n=100]vs 2.96%[n=20];11 .73%[n=80]vs 2.67%[n=18],respectively).Occupational exposure before and after intervention occurred most frequently when interns were preparing medicine,occupational exposure reporting rate in intervention group was higher than control group (72.97%[108/148]vs 50.11 % [221/441 ],χ2 =52.78,P <0.01 ).Conclusion The intensive training of occupational exposure protection among medical interns can improve the awareness and skills of occupational protec-tion,and reduce the occurrence of occupational exposure.