Identification and transcriptional activity analysis of core regulatory region of human guanylate binding protein 5 gene promoter / 中国生物制品学杂志

@#Objective To construct luciferase reporter plasmids of truncated fragments of different lengths of human guanylate binding protein 5(GBP5)gene promoter and analyze the transcriptional activity of each fragment to determine the core regulatory region.Methods GBP5promoter sequence was amplified by PCR,truncated into five fragments of different lengths and connected to pGL3-basic plasmid.The constructed recombinant plasmids pGL3-GBP5-11/21/31/41/51were transfected into 293FT cells and detected for luciferase activity.The binding sites of transcription factors in GBP5promoter region were predicted by JASPAR software,and Yin-Yang transcription factor 1(YY1)targeting the core regulatory region was selected and verified for the transcriptional regulatory activity.The CDS sequence of YY1 was amplified by PCR to construct the overexpression plasmid pIRES2-EGFP-YY1,which was then co-transfected to 293FT cells with plasmids pGL3-GBP5-21(-1 623 ～ +47 bp)and internal reference plasmid pRL-CMV,and detected for luciferase activity to analyze the regulation of transcription factor YY1 on GBP5 promoter activity.Results Colony PCR and double enzyme digestion identification proved that the plasmid of human GBP5 promoter reporter gene was correctly constructed;JASPAR software predicted that there were multiple transcription factor binding sites such as STAT1,YY1 and Foxp3 in GBP5promoter region.Double luciferase activity assay showed that pGL3-GBP5-21(-1 623 ～ +47 bp)showed the highest promoter activity,while the promoter activity of pGL3-GBP5-41(-520 ～ +47 bp)decreased significantly,suggesting that the core region of GBP5 promoter was located at upstream-1 623 ～-520 bp of 5 'UTR;Overexpression of YY1 significantly activated the GBP5 promoter activity and regulated the expression of GBP5.Conclusion The core regulatory region of human GBP5 promoter was located in upstream-1 623 ～-520 bp of the 5 'UTR,with a binding site of transcription factor YY1 existing in this region.Meanwhile,overexpression of YY1 significantly effected the activity of GBP5 promoter.

Analysis of 14 cases of melanosis caused by 1, 8-dinitronaphthalene and 1, 8-diaminonaphthalene / 中华劳动卫生职业病杂志

14 workers in the 1, 8-diaminonaphthalene workshop of a chemical company in Nantong City had symptoms or signs of varying degrees of pruritus and pigmentation of the face, neck and waist. Pathological examination of skin biopsies showed hyperkeratosis, the basal cells were liquefied and denatured. Seven workers were eventually diagnosed with occupational melanosis. To explore the causes of occupational melanosis caused by exposure to 1, 8-dinitronaphthalene and 1, 8-diaminonaphthalene, and to provide reference for the prevention and treatment of occupational melanosis in the future, this paper reported 14 cases of melanosis in the skin of workers in chemical industry.

Effect of Phospholipid Composition on Intradermal Delivery of Fluorescent Modified Cyclodextrin Liposomes / 中国药学杂志

OBJECTIVE: To compare the intradermal delivery effects of composite phospholipid liposomes composed of different proportions of soy phospholipids (SPC) and hydrogenated soy phospholipids (HSPC) on the fluorescent modified hydroxypropyl-β-cyclodextrin(HP-β-CD), and to optimize the phospholipid composition with the best skin retention. METHODS: The fluorescent probe, fluorescein isothiocyanate (FITC), was combined with HP-β-CD to prepare fluorescent modified cyclodextrin FITC-HP-β-CD. FITC-HP-β-CD was encapsulated in different composite phospholipid liposomes. The amount of the permeation in the receiving solution and skin retention of the cyclodextrin after 10 h were determined in the in vitro intradermal delivery experiment. RESULTS: The order of cyclodextrin permeation of liposomes in the receiving solution was SPC > S/H (3:1) > S/H (1:1) > S/H (1:3) > HSPC >FITC-HP-β-CD, while the order of cyclodextrin intradermal retention was S/H (1:1) > S/H (1:3) > HSPC > S/H (3:1) > SPC > FITC-HP-β-CD. CONCLUSION: Using SPC to prepare liposomes is more beneficial to promote the permeation of FITC-HP-β-CD into the skin than HSPC, but the addition of HSPC can increase the skin retention of FITC-HP-β-CD. The S/H(1:1) liposomes have the better intradermal delivery effect on the fluorescent modified cyclodextrin, of which the skin retention effect is the best.

Prevalence and Subtypes of Blastocystis in Alpacas, Vicugna pacos in Shanxi Province, China

Blastocystis, an enteric protist, has been reported to be an important cause of protozoal gastrointestinal manifestations in humans and animals worldwide. Animals harboring certain Blastocystis subtypes (STs) may serve as a potential source of human infection. However, information about the prevalence and genetic diversity of Blastocystis in alpacas is limited. In the present study, a total of 366 fecal samples from alpacas in Shanxi Province, northern China, were examined for Blastocystis by PCR amplification of the small subunit rRNA gene, followed by sequencing and phylogenetic analysis. The prevalence of Blastocystis in alpacas was 23.8%, and gender difference in the prevalence of Blastocystiswas observed. The most predominant Blastocystis ST was ST10, followed by ST14 and ST5. The detection of ST5, a potentially zoonotic genotype, indicates that alpacas harboring ST5 could be a potential source of human infection with Blastocystis. These data provide new insight into the prevalence and genetic diversity of Blastocystis in alpacas.

Correlation between nephropathy score and hormone threapy in minimal change disease / 临床与实验病理学杂志

Purpose To observe the clinical characteristics, expression of C4d and the morphology of podocyte lesions in steroid-sensitive minimal change disease (SS-MCD) ,steroidresistant minimal change disease (SR-MCD) and early focal segmental glomerulosclerosis (E-FSGS) ,as well as to analyze their differences among the three groups,and provide a novel method for effective evaluation the therapeutic effects of steroid and diagnosis of SR-MCD. Methods To study the clinical data from 24 cases of SS-MCD,30 cases of SR-MCD and 25 cases of E-FSGS as control,and all the biopsies were examined by light microscopy,immunohistochemistry and transmission electron microscopy. Meanwhile,the clinical characteristics,the morphology of podocyte lesion and the expression of C4d were observed. Results The average score of podocyte lesion of SR-MCD was higher than that of SS-MCD,but lower than that of E-FSGS (P< 0. 05) . C4d positive average score of SS-MCD was lower than that of both SR-MCD and E-FSGS (P < 0. 05) ,but there was no significant difference between SR-MCD and E-FSGS (P > 0. 05) . The sum of the average score of podocyte lesion and C4d positive average score of SS-MCD was lower than that of SRMCD and E-FSGS (P < 0. 01) ,however,there was also no significant difference between SR-MCD and E-FSGS(P > 0. 05) . The scores of IgM,C3d and C1q were not significantly different among the three groups. The area under the receiver operating curve (ROC) of the C4d positive score,podocyte lesion score and the sum of the two were 0. 753,0. 658 and 0. 803,respectively, and there was no significant difference between them and the optimal cutoff values were 3,1. 5,and 4. 5 points,respectively. Conclusions The C4d positive score,podocyte lesion score and the sum of the two scores of MCD (the last one is named for MCD nephropathy score in our study) can be used for evaluating the therapeutic effects of steroid and identification of SR-MCD,most especially MCD nephropathy score. The optimal cut-off values of the three kinds of scores are 3,1. 5,and 4. 5 points,respectively. When the values are exceeded,the clinicians should be reminded to follow-up and take appropriate treatment measures to patients.

Research progress of ocular manifestations and pathological mechanisms in Alport syndrome / 国际眼科杂志(Guoji Yanke Zazhi)

@#Alport syndrome(AS)is a genetic disease characterized by abnormal basement membrane structure of the kidneys, ears and eyes. The incidence of the disease is 1:5000 approximately. The report on ocular manifestations is relatively scarce, however, it is of great value to diagnosis of the disease. The ocular tissue histopathological analysis provides an effective method to uncover the pathological mechanisms of AS. Besides, it is good for understanding and treatment of AS.

Evaluation of chemical consistency of triterpene acids in ethanol extracts of Poria and acetic ether extracts thereof by UPLC-QTOF-MS/MS with full scan and mimic MRM mode / 药学学报

An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) method was developed to evaluate the chemical consistency of triterpene acids in ethanol extracts of Poria and acetic ether extracts thereof. First, high resolution mass spectrometry data were obtained with Full scan mode, by comparing with MS data from the reference compounds and literatures, a total of 23 components were unequivocally or tentatively identified in ethanol extracts and acetic ether extracts thereof. Then, a mimic multiple reaction monitoring (mMRM) mode was established using UPLC-QTOF-MS/MS to quantify the triterpene acids in ethanol extracts and acetic ether extracts thereof. Eleven components were absolutely quantified with reference compounds, while 12 components without reference compounds were relatively quantified with peak areas, the transfer and enrichment rate of triterpene acids during liquid-liquid extraction were calculated. It was found all of the 23 triterpene acids identified in Poria ethanol extracts could be transferred into acetic ether extracts with high transfer and enrichment rate. The present study provides not only scientific evidence for further extraction of triterpene acids in Poria by acetic ether, but also an approach for comprehensive evaluation of the chemical consistency of herbal medicine extracts before and after the liquid-liquid extraction.

Differences of brain pathological changes and cognitive function after bilateral common carotid artery occlusion between Sprague-Dawley and Wistar rats / 生理学报

The aim of the present study was to investigate the differences of the pathological changes and cognitive function after bilateral common carotid artery occlusion (BCCAO) between Sprague-Dawley (SD) and Wistar rats. Male SD and Wistar rats were randomly divided into 2 groups, respectively: sham operated (S-sham and W-sham) and operated (S-BCCAO and W-BCCAO) groups. The survival rate and the rate of loss of pupillary light reflex (PLR) were observed on day 1, 3, 7, 14 and 28 after the operation, and the light-dark box, Y-maze and odor recognition tests were performed to detect cognitive function on day 28 after the operation. HE and Luxol fast blue staining were used to observe the pathological changes of gray matter (hippocampus), white matter (optical tract), optic nerve, and retina. The results showed that the survival rate of the W-BCCAO group was 62.5%, and PLR loss rate was 100%; whereas the survival rate of the S-BCCAO group was 100%, and PLR loss rate was 58.3%. In the W-BCCAO group, percentages of time spent and distance traveled in the light box were more than those in the W-sham group, but there was no statistical significance between the S-BCCAO and S-sham groups. In the S-BCCAO group, the percentages of time spent and distance traveled in the III arm (labyrinth arm) of the Y-maze were less than those in the S-sham group, but no statistical significance was found between the W-BCCAO group and W-sham group. In the S-BCCAO group, the discrimination ratio of the odor recognition task was less than that in the S-sham group, but no statistical significance could be seen between the W-BCCAO and W-sham groups. Ischemic injury was observed in the CA1 area of the hippocampus in the S-BCCAO group, but no readily visible damage was observed in the W-BCCAO group. Ischemic injury of the visual beam and optic nerve was observed in both the S-BCCAO and W-BCCAO groups. Compared with the corresponding sham groups, the S-BCCAO and W-BCCAO groups showed serious retinal damage with significant thinner retina. The ganglion cell layer (GCL), inner plexiform layer (IPL), and outer plexiform layer (OPL) were thinner in the S-BCCAO group, but no statistical significances were shown in the other layers. All the layers, except the outer nuclear layer (ONL), were significantly thinner in the W-BCCAO group. The results indicate that there are differences of the pathological changes in the hippocampus and visual conduction pathway after BCCAO between SD and Wistar rats, and the degree of learning and memory injury was also different, which suggests that the vascular dementia model of different rat strains should be selected according to research purpose.

Comparative study of penetration-enhancing effect in vitro of cinnamon oil and cinnamaldehyde on ibuprofen / 中国中药杂志

To compare the penetration-enhancing effect of cinnamon oil and its main components (cinnamaldehyde) on ibuprofen and their self-percutaneous absorption behavior in vitro. Firstly, cinnamon oil was extracted by steam distillation, then the compositions were analyzed by gas chromatography mass spectrometry (GC-MS) and the cinnamaldehyde content in cinnamon oil was determined by high performance liquid chromatography (HPLC). With azone as positive control, ibuprofen as model drug, cinnamon oil and cinnamaldehyde as penetration enhancers (PE) were prepared and administered to the SD rat's abdominal skin. The penetration-enhancing effects of cinnamon oil and cinnamaldehyde and their own transdermal absorption properties were compared. The results showed that yield of cinnamon oil was (3.55±0.36)% (=3), and the cinnamaldehyde content in cinnamon oil was (73.48±0.21)% (=3). As compared with blank group, the enhancing rate (ER) of cinnamon oil, cinnamaldehyde, and azone was 3.56, 1.13, 2.47 respectively. The cumulative penetration rate of cinnamaldehyde in cinnamon oil and cinnamaldehyde monomer in 24 h was (63.30±0.98)%, (51.03±3.34)% (=4) respectively. The penetration-enhancing effect of cinnamon oil was significantly better than that of cinnamaldehyde, indicating the existence of muti-component synergy. The penetration rate of cinnamaldehyde in cinnamon oil was higher than that of cinnamaldehyde monomer, suggesting that a "pull effect" may be present.

Neuroanatomical Circuitry between Kidney and Rostral Elements of Brain: a Virally Mediated Transsynaptic Tracing Study in Mice / 华中科技大学学报（医学）（英德文版）

The identity of higher-order neurons and circuits playing an associative role to control renal function is notwell understood.We identified specific neural populations of rostral elements of brain regions that project multisynaptically to the kidneys in 3~ days after injecting a retrograde tracer pseudorabies virus (PRV)-614 into kidney of 13 adult male C57BL/6J strain mice.PRV-614 infected neurons were detected in a number of mesencephalic (e.g.central amygdala nucleus),telencephalic regions and motor cortex.These divisions included the preoptic area (POA),dorsomedial hypothalamus (DMH),lateral hypothalamus,arcuate nucleus (Arc),suprachiasmatic nucleus (SCN),periventricular hypothalamus (PeH),and rostral and caudal subdivision of the paraventricular nucleus of the hypothalamus (PVN).PRV-614/Tyrosine hydroxylase (TH) double-labeled cells were found within DMH,Arc,SCN,PeH,PVN,the anterodorsal and medial POA.A subset of neurons in PVN that participated in regulating sympathetic outflow to kidney was catecholaminergic or serotonergic.PRV-614 infected neurons within the PVN also contained arginine vasopressin or oxytocin.These data demonstrate the rostral elements of brain innervate the kidney by the neuroanatomical circuitry.

Cement leakage factors in clinical vertebroplasty / 中国组织工程研究

BACKGROUND: Vertebroplasty (PVP) and kyphoplasty (PKP) are important methods for the treatment of osteoporotic vertebral compression fractures in the elderly. Although bone cement has certain liquidity and vertebral fractures are often in different situations, bone cement leakage rate is still high. OBJECTIVE: Based on the theoretical discussion and clinical analysis, to study the theoretical causes of bone cement leakage and effective prevention methods. METHODS: A total of 162 cases (186 vertebrae) were treated with three methods of vertebroplasty. Group A: 64 cases with 78 vertebrae were treated with conventional cemented vertebroplasty; Group B: 57 cases with 65 vertebrae were treated with cemented vertebroplasty using cement pump; Group C: 41 cases with 43 vertebrae were treated with balloon kyphoplasty. The leakage of bone cement was observed in three groups. RESULTS AND CONCLUSION: Of the 186 vertebrae, postoperative bone cement leakage occurred in 25 vertebrae, with the leakage rate of 13% (25/186). Group A had bone cement leakage in 11 vertebrae, and the leakage rate was 14% (11/78). Group B had bone cement leakage in 8 vertebrae, and the leakage rate was 12% (8/65). Group C had bone cement leakage in 6 vertebrae, and the leakage rate was 14% (6/43). There was no significant difference among the three groups in the leakage rate of bone cement (P > 0.05). That is to say, the causes of bone cement leakage are not completely controllable, and the leakage position has some randomness. Strict and careful imaging monitoring is an intuitive method to prevent bone cement leakage.

Cement leakage factors in clinical vertebroplasty / 中国组织工程研究

BACKGROUND: Vertebroplasty (PVP) and kyphoplasty (PKP) are important methods for the treatment of osteoporotic vertebral compression fractures in the elderly. Although bone cement has certain liquidity and vertebral fractures are often in different situations, bone cement leakage rate is still high. OBJECTIVE: Based on the theoretical discussion and clinical analysis, to study the theoretical causes of bone cement leakage and effective prevention methods. METHODS: A total of 162 cases (186 vertebrae) were treated with three methods of vertebroplasty. Group A: 64 cases with 78 vertebrae were treated with conventional cemented vertebroplasty; Group B: 57 cases with 65 vertebrae were treated with cemented vertebroplasty using cement pump; Group C: 41 cases with 43 vertebrae were treated with balloon kyphoplasty. The leakage of bone cement was observed in three groups. RESULTS AND CONCLUSION: Of the 186 vertebrae, postoperative bone cement leakage occurred in 25 vertebrae, with the leakage rate of 13% (25/186). Group A had bone cement leakage in 11 vertebrae, and the leakage rate was 14% (11/78). Group B had bone cement leakage in 8 vertebrae, and the leakage rate was 12% (8/65). Group C had bone cement leakage in 6 vertebrae, and the leakage rate was 14% (6/43). There was no significant difference among the three groups in the leakage rate of bone cement (P > 0.05). That is to say, the causes of bone cement leakage are not completely controllable, and the leakage position has some randomness. Strict and careful imaging monitoring is an intuitive method to prevent bone cement leakage.

Effect of bone morphogenetic protein-2 on human heart valve interstitial cells calcification in vitro / 中华心血管病杂志

<p><b>OBJECTIVE</b>To establish human heart valve interstitial cells calcification culture model in vitro, and observe the effect of bone morphogenetic protein-2 (BMP-2) on calcification of human heart valve interstitial cells.</p><p><b>METHODS</b>Human heart valve interstitial cells were cultured in vitro, and divided into control group: cells were cultured in conventional media plus recombinant human BMP-2 treatment and experimental group: besides above treaments, calcification inducers ( recombinant human BMP-2, β-glycerophosphate, L-ascorbic acid, dexamethasone) were added to the culture media. The two group of cells were cultured for 14 days and were stained by Von Kossa, then the cell calcification was observed in this valvular interstitial cells calcification culture model in vitro. Protein expression of intercellular adhesion molecule 1 (ICAM-1), interleukin 8, BMP-2 and BMP-4 was determined by Western blot and BMP-2 secretion was measured by ELISA.</p><p><b>RESULTS</b>In the control group, the structure of human heart valve interstitial cells was clear, and the spindle and radial growth shaped cellular morphology was visible, and Von Kossa staining was negative. In the experimental group, the nuclei become darker in color, and granular sediment distribution was seen surrounding cells, and Von Kossa staining was positive, the cells were forming nodules of calcification. The protein expression of ICAM-1, interleukin 8, BMP-2 and BMP-4 in the experimental was significantly higher than that of the control group (all P < 0.05). The expression of BMP-2 in the experimental group was also significantly higher than that in control group ((92.5 ± 4.9) pg/ml vs. (22.2 ± 1.9) pg/ml, P < 0.05).</p><p><b>CONCLUSION</b>Human BMP-2, β-glycerophosphate, L-ascorbic acid, and dexamethasone can induce human heart valve interstitial cells calcification and enhance inflammation in vitro by stimulating the secretion of BMP-2.</p>

Optimization of the whole-cell catalytic activity of recombinant Escherichia coli cells with surface-immobilized organophosphorus hydrolase.

Previous studies have verified the feasibility of using Escherichia coli systems that display organophosphorous hydrolase (OPH) on the cell surface as whole-cell catalysts. However, the inefficient display of the enzyme on cell surfaces remains unaddressed. In the present study, multiple optimization experiments on full-length and truncated ice nucleation protein anchors, E. coli host cells, culture media, and culture conditions were performed to optimize whole-cell OPH enzymatic activity. The results show that apart from the dramatic effect of isopropyl-b-d-thiogalactoside concentration and culture temperature, the coordination between the anchor protein, culture media, and host cells is essential for highly efficient OPH display. Under optimal conditions, namely, culturing in M9 medium, 20 °C induction temperature, 0.1 mmol l-1 IPTG, and 100 μmol l-1 Co2+, the engineered E. coli strain MB109-406 that expresses the fusion enzyme InaK-N-OPH exhibited a whole-cell OPH activity of 0.62 U mg-1 ?cell d.wt. This result is much higher than that of several currently available OPH-displaying systems, which shows the potential of the current system for further large-scale industrial or environmental applications.

Determination of Lipoprotein Lipase and Hepatic Lipase in Children with Nephrotic Syndrome / 实用儿科临床杂志

Objective To investigate pathogenesis of liprd metabolism disorder in children with nephrotic syndrome. Methods Serum lipid and plasma llpoprotein lipase and hepatic lipase were detected in 62 nephrotic syndrome children and 30 normal children, respectively. Results The activity of lipoprotein lipase and hepatic lipase was lower than that in normal control group, while serum cholesterol, triglycerides and low -density lipoprotein in nephrotic group were higher than those in control group. Lipoprotein lipase and hepatic lipase were negative correlation with triglycerides and low - density lipoprotein, respectively. Conclusions Reduced activity of lipoprotein lipase and hepatic lipase is one of causes leading to hypertriglyceridemia in nephrotic syndrome.