Study on method of osthole loaded by exosomes / 中国药理学通报

Aim To construct a drug delivery system of osthole loaded by exosomes. Methods Osthole could inhibit the proliferation of ovarian cancer cells by literature. SKOV3 cells were treated with 80 (µnol • L

Dahuang zhechong pills regulate ASICla/VEGF pathway and alleviate hepatic fibrosis / 中国药理学通报

Aim To examine the therapeutic effects of DHZCP on carbon tetrachloride(CCl4)-induced chemical hepatic fibrosis model in rats and the mechanism of acid-sensitive ion channels 1a(ASIC1a)and vascular endothelial growth factor(VEGF)-related mechanisms.Methods The rats were injected intraperitoneally with CCl4 vegetable oil mixture to establish hepatic fibrosis model,and randomly divided into six groups:control group,hepatic fibrosis model group,DHZCP low dose group,DHZCP medium dose group,DHZCP high dose group and colchicine(Col)positive control group.HE staining was used to observe the pathological changes of hepatic structures in each group,Masson staining to view the production of collagen fibers in each group,and immunohistochemistry,Western blot,q-PCR to investigate the expression level of ASIC1a,CaMKKβ,VEGF,α-SMA,Collagen-I proteins.Results In model group,serum ALT and AST levels were obviously up-regulated,liver tissue structure was severely damaged,and ASIC1a,CaMKKβ,VEGF,α-SMA,Collagen-I gene and protein expression levels were significantly elevated.Compared with model group,each treatment group of DHZCP could markedly alleviate the pathological changes of liver fibrosis caused by CCl4,significantly reduce the serum ALT and AST levels,and dose-dependently down-regulate the gene and protein expression levels of ASIC1a,CaMKKβ,VEGF,α-SMA,Collagen-I,etc.Conclusions DHZCP ameliorates hepatic fibrosis in rats,and its mechanism of action may be associated with the regulation of ASIC1a/VEGF.

Applications and challenges of metagenomic next-generation sequencing in the detection of 2019 novel coronavirus / 中华检验医学杂志

As one of the two methods for 2019 novel coronavirus (2019-nCoV), gene sequencing is different from quantitative real-time PCR (RT-PCR) in detection principles. Therefore, gene sequencing has its own pros and cons in clinical application. Currently, metagenomic next-generation sequencing (mNGS) is the most commonly used technology in clinical application.Due to its broad coverage of all types of pathogens, mNGS demonstrates incomparable advantage in rapid identification of novel pathogens such as 2019-nCoV. In addition, it can simultaneously identify other pathogens except 2019-nCoV and mixed infections. On the other hand, however, due to the complexity of mNGS and long detection time, it is unlikely to achieve the purpose of wide-range and rapid diagnosis of 2019 n-CoV. Therefore, mNGS can complement RT-PCR to achieve best clinical application.

Understanding the influence factors in viral nucleic acid test of 2019 novel coronavirus / 中华检验医学杂志

At present, the prevention and control of new coronavirus has entered a critical period. However, the use of quantitative real-time PCR (qRT-PCR)assays for the detection of viral nucleic acid, as a crucial diagnostic approach, has been doubted in clinical practice. Herein, we have reviewed the current status of epidemic prevention and control, latest development of detection technologies, disease characteristics, clinical sampling and transport. It has also discussed the factors that may affect the performance of viral nucleic acid detection, and suggested some effective methods to improve the detection performance of the assays.

Advantages and challenges of metagenomic next-generation sequencing (mNGS) in the detection of 2019 novel coronavirus / 中华检验医学杂志

As one of the two methods for 2019 novel coronavirus (2019-nCoV), gene sequencing is different from quantitative real-time PCR (RT-PCR) in detection principles. Therefore, gene sequencing has its own pros and cons in clinical application. Currently, metagenomic next-generation sequencing (mNGS) is the most commonly used technology in clinical application. Due to its broad coverage of all types of pathogens, mNGS demonstrates incomparable advantage in rapid identification of novel pathogens such as 2019-nCoV. In addition, it can simultaneously identify other pathogens except 2019-nCoV and mixed infections. On the other hand, however, due to the complexity of mNGS and long detection time, it is unlikely to achieve the purpose of wide-range and rapid diagnosis of 2019 n-CoV. Therefore, mNGS can complement RT-PCR to achieve best clinical application.

Correlation analysis of pediatric sepsis and intestinal flora imbalance / 中国小儿急救医学

Objective To investigate the interaction between the gut microbiota and the sepsis in children by comparing the difference of gut microbiota between septic children and healthy children. Methods Genome was extracted from excrements of 18 cases of sepsis and 6 cases of healthy children. After genom-ic extraction,the hypervariable region of 16S rDNA gene were amplified and a small fragment library was constructed,and high-throughput sequencing was carried out,then the data of the lower machine was effec-tively sequenced by biological information processing. We could seek for the species that had changed signifi-cantly due to sepsis by comparing the diversity and the differences in the composition of intestinal flora between the two groups. Results The gut microbiome of the sepsis group was distinct from that of the health group. The operational taxonomic units in the sepsis group were significantly reduced compared with healthy group(P＝0. 001). The gut microbiome of children with sepsis had significantly lower diversity and richness compared with healthy group(P<0. 05). A total of 7 species were shown to be differentially abundant be-tween septic patients and healthy controls. The genus Pseudomonadales,Carnobacteriaceae and Granulicatella elegans were significantly more abundant in the sepsis group; meanwhile the genus Pasteurellaceae, Ruminococcus,Lactobacillus rogosae and Anaerostipes butyratucus were less abundant in the sepsis group. In addition,the Granulicatella elegans was characteristically present in the intestine of children with sepsis(P<0. 001). Conclusion The microbial diversity and structure of the gut microbiome in children with sepsis are significantly different from those of healthy children. Our data suggest biomarkers identified in this study might participate in the pathogenesis or development process of sepsis.

Chromosome polymorphisms and their influence on semen quality and sperm DNA integrity in males undergoing IVF/ICSI / 中华男科学杂志

Objective@#To investigate the incidence of chromosome polymorphisms and their influence on semen quality and sperm DNA integrity in male patients receiving in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI).@*METHODS@#We retrospectively analyzed the chromosomal karyotypes and the types and incidence rate of chromosome polymorphisms in 2 370 male patients undergoing IVF/ICSI between June 2016 and June 2018. We classified the patients into groups A (with variation in the secondary constriction region in the autosomal long arm), B (with variation in the short arm of the D/G group chromosomes), C (with interbrachial inversion of chromosome 9) and D (with Y chromosome polymorphisms), and compared the semen parameters and sperm DNA fragmentation indexes (DFI) between the patients with chromosome polymorphisms and those with normal chromosomes.@*RESULTS@#Totally, 154 (6.50%) of the patients undergoing IVF/ICSI were found with chromosome polymorphisms, including 34 cases of secondary constriction variation in the long arm of the autosome (1.43% ［34/2 370］, 22.08% ［34/154］), 82 cases of short arm polymorphisms of the D/G group chromosomes (3.46% ［82/2 370］, 53.25% ［82/154］), 26 cases of interbrachial inversion of chromosome 9 (1.10% ［26/2 370］, 16.88% ［26/154］), 10 cases of Y chromosome polymorphisms (0.42% ［10/2 370］, 6.50% ［10/154］), and 2 cases of mixed chromosome polymorphisms (0.08% ［2/2 370］, 1.42% ［2/154］). The total sperm count was lower in group D than in the other polymorphism groups and the normal chromosome group, but with no statistically significant difference among the five groups (P > 0.05). The sperm progressive motility was also lower in group D than in the other five groups, with statistically significant difference from group B (27.5 ± 13.5 vs. 41.5 ± 21.1, P = 0.027), but not from the other groups (P > 0.05). No statistically significant difference was observed in the sperm DFI between the polymorphism groups and the normal chromosome group (P > 0.05), or among the polymorphism groups (P > 0.05). The proportion of normal semen was lower in group D than in the other four groups, but with no statistically significant difference among the five groups (P > 0.05). The incidence rate of asthenospermia was higher in group D than in the other four groups, but with no statistically significant difference among the five groups (P > 0.05), and so was that of oligoasthenospermia, with statistically significant difference from the normal chromosome group (30.0% vs 8.0%, P = 0.041), but not from the other polymorphism groups (P > 0.05).@*CONCLUSIONS@#Short arm polymorphisms of the D/G group chromosomes are the most common type of chromosome polymorphisms in male patients undergoing IVF/ICSI. Polymorphisms of the Y chromosome have a negative effect on semen quality, while those of the other chromosomes do not significantly affect semen quality and sperm DNA integrity.

Correlation analysis of pediatric sepsis and intestinal flora imbalance / 中国小儿急救医学

Objective@#To investigate the interaction between the gut microbiota and the sepsis in children by comparing the difference of gut microbiota between septic children and healthy children.@*Methods@#Genome was extracted from excrements of 18 cases of sepsis and 6 cases of healthy children.After genomic extraction, the hypervariable region of 16S rDNA gene were amplified and a small fragment library was constructed, and high-throughput sequencing was carried out, then the data of the lower machine was effectively sequenced by biological information processing.We could seek for the species that had changed significantly due to sepsis by comparing the diversity and the differences in the composition of intestinal flora between the two groups.@*Results@#The gut microbiome of the sepsis group was distinct from that of the health group.The operational taxonomic units in the sepsis group were significantly reduced compared with healthy group(P=0.001). The gut microbiome of children with sepsis had significantly lower diversity and richness compared with healthy group(P<0.05). A total of 7 species were shown to be differentially abundant between septic patients and healthy controls.The genus Pseudomonadales, Carnobacteriaceae and Granulicatella_elegans were significantly more abundant in the sepsis group; meanwhile the genus Pasteurellaceae, Ruminococcus, Lactobacillus_rogosae and Anaerostipes_butyratucus were less abundant in the sepsis group.In addition, the Granulicatella_elegans was characteristically present in the intestine of children with sepsis(P<0.001).@*Conclusion@#The microbial diversity and structure of the gut microbiome in children with sepsis are significantly different from those of healthy children.Our data suggest biomarkers identified in this study might participate in the pathogenesis or development process of sepsis.

Adenanthin Induces Differentiation of Acute Promyelocytic Leukemia Cells by Targeting Peroxiredoxin III / 中国实验血液学杂志

OBJECTIVE@#To investigate the differentiation of acute promyelocytic leukemia (APL) cells induced by adenosine targeting Prx III.@*METHODS@#HL-60 cells were divided into four groups: control group, all-trans retinoic acid (ATRA) group, adenanthin group and ATRA+adenanthin group. Cell morphologic changes were observed under optical microscope. The influence of adenanthin on the differentiation of HL-60 was observed by nitro blue tetrazolium chloride (NBT) test. Cell surface differentiation antigens CD11b expression was measured by flow cytometry. The protein expression of Prx III was detected by immunohistochemical assay.@*RESULTS@#Adenanthin could induce the differentiation of HL-60 cells; the NBT reduction positive rate in ATRA+adenanthin group was significantly higher than that in ATRA group and adenanthin group (P＜0.05). The percentage of CD11b positive cells in ATRA+adenanthin group (43.62％±1.38％) was higher than that in adenanthin group (28.15％±1.78％), ATRA group (36.72％±1.33％) and control group (7.99％±1.78％) (P＜0. 05). The content of Prx Ⅲ protein in adenanthin group was significantly higher than that in control group and ATRA group (P＜0.05).@*CONCLUSION@#Adenanthin and ATRA have a synergistic effect on the differentiation and maturation of HL-60 cells, and its mechanism may be related with regulation of Prx III expression.

Effect of Helicobacter pylori infection on gut microbiota in children / 中国小儿急救医学

Objective To investigate the effects of Helicobacter pylori( HP) on gut microbiota in children by comparing the difference of gastric microbiota between HP-positive and HP-negative individuals. Methods Genome was extracted from excrements of 8 HP-positive cases and 8 HP-negative cases. After genomic extraction,the hypervariable region of 16S rRNA gene were amplified and a small fragment library was constructed,and high-throughput sequencing was carried out, then the data of the lower machine was effectively sequenced by biological information processing. We could seek for the species that have changed significantly due to HP infection by comparing the differences in the composition of intestinal flora between the two groups. Results Compared with HP-negative group,HP-positive group showed less OTUs. The dif-fenece of biodiversity between them was conspicuous. The Caproiciproducens,Enterobacteriaceae,Enterobac-teriales,Blautia-obeum,Esherichia-albertli,human-gut-metagenome and Dorea in HP-positive group were sig-nificantly higher than HP-negative group,while the Bacteroides-uniformis, Bacteroidaceae and Bacteroides in HP-negative group were significantly higher than HP-positive group. Conclusion HP could significantly affect the structure and composition of gut microbiota in children.

Clinical features and etiological analysis of suspected pertussis in infants / 中国小儿急救医学

Objective To investigate the clinical features and pathogenic types of suspected pertussis syndrome in infants,so as to provide reference for the treatment. Methods Seventy-one infants of suspected pertussis in Shanghai Children′s Medical Center from Nov 2016 to Aug 2017 were detected by the Filmarray which can detect Bordetella pertussis and 17 viruses. According to the results,the infants were divided into two groups:pertussis group(n=29) and pertussis-like group(n=42). According to the severity of the dis-ease,they were divided into mild group(n=50) and severe group(n=21). Clinical data was retrospectively analyzed and compared. Results All 71 infants came to the hospital with cough. Paroxysmal cough happened in 18 cases(62. 1％)in pertussis group,more common than that in pertussis-like group[9 cases(21. 4％)] (χ2 =12. 023,P<0. 01),and the WBC count,lymphocyte ratio,the mixed virus infection rate were higher in pertussis group than those in pertussis-like group[(20. 00 ± 8. 62) × 109/L vs. (13. 42 ± 6. 58) × 109/L,t=-3. 647,P<0. 01;(70. 38 ± 8. 97)％ vs. (56. 26 ± 20. 38)％,t = -3. 967,P <0. 01;22 cases(75. 9％) vs. 16 cases(38. 1％),χ2 =9. 836,P<0. 01]. The cases of mixed bacterial infection in pertussis-like group were 13(31. 0％),which was higher than that in pertussis group[3(10. 3％)](χ2 =4. 173,P<0. 05). The incidence of cyanosis was found in 12 cases(57. 1％)in severe group,which was more common than that of mild group[12 cases(24. 0％)](χ2 =7. 260,P<0. 01), and hospitalization days were(14. 5 ± 7. 8) days, which was higher than that in mild group[(7. 0 ± 3. 1)days] (t= -4. 250, P<0. 01). The infants in the pertussis group were given macrolides antibiotics and sulfamethoxazole complex,and the infants in the pertus-sis-like group were treated with antiviral and other specific treatment. Among 71 infants, 67 cases (94. 4％) were cured and 3 cases (4. 2％) were improved. Conclusion The clinical features of suspected pertussis in infants are not typical, so the early pathogenic diagnosis is very important. Filmarray detection system for multi PCR system can detect 20 kinds of pathogens with short operation time,which is very helpful for the early and rapid diagnosis of pathogens and rational use of drugs. It is worthy of clinical promotion.

Clinical application of FilmArray respiratory panel in Mycoplasma pneumoniae detection / 中华检验医学杂志

Objective To evaluate the clinical efficacy and application value of FilmArray Respiratory Panel(FilmArray RP)for Mycoplasma pneumoniae(MP)detection.Methods Methodological evaluation.A total of 360 nasopharyngeal swabs or sputum samples were collected from hospitalized children with community acquired pneumonia in Shanghai Children′s Medical Center,Shanghai Jiao Tong University School of Medicine(SCMC)from November 2016 to May 2017.Among them, there were 197 males and 163 females.There were 185 cases in 0-2 years old group,116 cases in 3-6 years old group and 59 cases in 7-16 years old group.FilmArray RP and serological test were respectively used to detect MP infection. Polymerase chain reaction(PCR)was also performed on all the positive samples of MP detected by FilmArray RP,negative samples of FilmArray RP but positive by serological test and some negative samples of the two test methods selected by simple random sample.The results of FilmArray RP and serological test,the positive rates of different age groups detected by FilmArray RP and the rates of MP combined with virus were statistically analyzed by χ2test.Results Among 360 patients with community acquired pneumonia,39 (10.83%,39/360)were MP positive detected by FilmArray RP.In contrast,93 patients(25.83%,93/360)were diagnosed as MP infections using serological test.Results from the two methods were statistically significant(χ2=27.05, P=0.00).Among them, 39 cases of MP detected by FilmArray RP were also positive by PCR.Sixty-one negative samples of FilmArray RP but positive by serological test and 22 negative samples of the two test methods selected by simple random sample were also negative by PCR.Among the positive samples detected by FilmArray RP,the positive rate of patients aged 7-16 years(28.81%,17/59) was higher than that of patients aged 0-2 years(5.95%,11/185)and patients aged 3-6 years(9.48%,11/116).The positive rates of different age groups were statistically significant(χ2=24.54, P=0.00).The proportion of MP combined with virus infection in 0-2 years old group(81.82%,9/11)was higher than that in 3-6 years old group(45.45%,5/11)and 7-16 years old group(29.41%,5/17),the difference was statistically significant(χ2=7.41, P=0.03).Conclusions FilmArray RP is a rapid method in MP detection.It has great advantages in the detection of MP and virus mixed infection.

Xuebijing Injection () and Resolvin D1 Synergize Regulate Leukocyte Adhesion and Improve Survival Rate in Mice with Sepsis-Induced Lung Injury / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of combined application of Xuebijing Injection ( , XBJ) and resolvin D1 (RvD1) on survival rate and the underlying mechanisms in mice with sepsisinduced lung injury.</p><p><b>METHODS</b>The cecal ligation and puncture (CLP) method was used to develop a mouse sepsis model. Specific pathogen free male C57BL/6 mice were randomly divided into 5 groups (n=20 each): sham, CLP, CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1. After surgery, mice in the CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1 groups were given XBJ (25 μL/g body weight), RvD1 (10 ng/g body weight), and their combination (the same dose of XBJ and RvD1), respectively. In each group, 12 mice were used to observe 1-week survival rate, while the rest were executed at 12 h. Whole blood was collected for flow cytometric analysis of leukocyte adhesion molecules CD18, lung tissues were harvested for observing pathological changes, and testing the activity of myeloperoxidase (MPO) and the expression of intercellular cell adhesion molecule 1 (ICAM-1).</p><p><b>RESULTS</b>Compared with the CLP group, the histopathological damage of the lung tissues was mitigated, MPO activity was decreased in the CLP+XBJ and CLP+RvD1 groups (P<0.05). In addition, the 1-week survival rate was improved, proportion of CD18-expressing cells in whole blood and ICAM-1 protein expression in lung tissue were decreased in the CLP+XBJ+RvD1 group (P<0.05 or P<0.01).</p><p><b>CONCLUSIONS</b>XBJ together with RvD1 could effectively inhibit leukocyte adhesion, reduce lung injury, and improve the survival rate of mice with sepsis.</p>

Study of endothelial cell toxicity of low dosage of extracellular histones / 临床儿科杂志

Objective To investigate the effects of low dose of extracellular histone on endothelial cells in infectious diseases such as sepsis. Methods The endothelial cells were treated with 10 μg/mL recombinant human histone H3/H4 complex in replacement of calf thymus histones (CTH) for various periods of time, and the morphology changes and the viability of the endothelial cells were recorded. In addition, flow cytometry was applied to identify the characteristics of endothelial cells and enzyme-linked immunosorbnent assay (ELISA) was used to detect the extracellular histones level in endothelial cells culture. Results The low dose of CTH could continuously induce endothelial cells death, cell morphological changes and function loss, which was reproduced by 10 μg/mL recombinant histone H3/H4 complex. Results of histones quantitation showed that histone can cause a series of intracellular reactions in a short period of time. Conclusions It is showed that 10 μg/mL H3/H4 can induce the toxicity in infectious disease and this level of the dose is a lower than those used in previous studies and more close to the pathological conditions.

Cytotoxicity of extracellular histones on vascular endothelial cells and the underlying mechanisms in sepsis / 中国小儿急救医学

Objective To investigate the change of extracellular histone level as well as the mecha-nism of the cytotoxicity of extracellular histones on vascular endothelial cell in sepsis. Methods Septic chil-dren admitted to PICU in Shanghai Children′s Medical Center between January 2010 and December 2014 were included in the present study. According to the diagnostic criteria of sepsis,the patients were divided into the sepsis group(51 cases) and the severe sepsis group(79 cases),with healthy children as the control group (108 cases). Patients in the severe sepsis group were further divided into the survival group(45 cases) and the non-survival group ( 34 cases ) based on 28-day mortality. The plasma concentration of extracellular histones in these children was determined and its correlation with the severity of sepsis was analyzed. Human umbilical vein endothelial cells(HUVECs) were incubated with calf thymus histone(CTH) at various con-centrations(0,50,100,200 and 300μg/ml) or different time periods(200μg/ml,0,5,15,30,45 and 60 mi-nutes) . The treated cells were subject to flow cytometer to measure the cell survival rate and scanning/trans-mission electron microscopy to observe their morphological changes. Western blot was used to detect the ex-pression of IκB and phosphor-p38/p38 in nuclear factor ( NF )-κB and mitogen-activated protein kinase ( MAPK) signaling pathways,while ELISA was used to determine the levels of tumor necrosis factor-α and interleukin-6. Results The levels of circulating histones in the septic children(2. 29 ± 1. 00) and severe sep-tic children ( 19. 17 ± 10. 20 ) were significantly higher than that of healthy controls ( 0. 23 ± 0. 26 ) ( P <0. 001),and the histone levels in the severe septic children were even higher(P<0. 001). Among the chil-dren diagnosed as severe sepsis,the level of circulating histones in the non-survivors was significantly higher than that in the survivors(29. 47 ± 5. 99 vs. 10. 94 ± 2. 68,P<0. 001). The survival rate of HUVEC gradually decreased along with the increase of CTH concentration or the treatment period in vitro. Data from electron microscopy showed that CTH treatment could directly disrupt the plasma membrane of HUVEC. Histones could also activate NF-κB and MAPK pathways,leading to the release of large amount of tumor necrosis fac-tor-α and interleukin-6. Conclusion The levels of extracellular histones in the septic children are correlated with the severity of sepsis. CTH can induce HUVEC death in a dose-and time-dependent manner. Extracellu-lar histone-induced endothelial dysfunction may mediate the progression of sepsis and such cytotoxicity might be due to the destruction of endothelial cell membranes and activation of the NF-κB and MAPK signaling pathways.

Expression and purification of recombinant human Cosmc extracellular domain in Sf-9 insect cells / 中国免疫学杂志

Objective:In the present study,Bac-to-Bac baculovirus expression system was used to obtain recombinant human Cosmc extracellular domain protein,which can lay the foundation for the research about the structure and function of Cosmc protein in vitro,and simultaneously provide ideas for the research of O-glycosylation and related diseases. Methods: The Cosmc extracellular domain ( Cosmc-ED) gene was cloned into a transfer vector pFastBac1 to form the recombinant donor plasmid pFastBac1-Cosmc ED, which was transformed into competent cells DH10Bac. By using blue-white selection and PCR analysis,we could obtain recombinant shuttle vector rBacmid-Cosmc ED. Then, the recombinant gene DNA of rBacmid-Cosmc ED was used to transfect Sf-9 mediated by cationic lipid formulation,and the recombinant baculovirus bacmid was obtained,which was further used to infect the serum-free cell Sf-9 to express Cosmc-ED in the supernatant. Then the protein of interest was detected by SDS-PAGE and Western blot and purified with Ni-NTA affinity column. Results:SDS-PAGE and Western blot analysis showed a specific band about 33 kD,consistent with the interest protein. Mass spectrometry results further prove that the protein was Cosmc extracellular domain protein. Conclusion: Human Cosmc-ED protein can be successfully expressed in Sf-9 insect cells and laid basis for subsequent studies.

Association of Cosmc gene mutation with susceptibility to Henoch-Schönlein purpura in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the presence of Cosmc gene mutation in children with Henoch-Schönlein purpura (HSP) and the association between Cosmc gene mutation and the susceptibility to HSP.</p><p><b>MESULTS</b>Eighty-four children who were diagnosed with HSP between March 2014 and December 2015 were selected as the HSP group. Fifty-eight healthy volunteers matched for age and sex were enrolled as the control group. Fasting venous blood (5 mL) from the two groups was collected in EDTA anticoagulated tubes, followed by the isolation of peripheral blood mononuclear cells (PBMCs) through density gradient centrifugation. Genomic DNA was extracted from PBMCs according to the manufacturer's protocol, and the whole exon region of Cosmc gene was amplified by touch-down polymerase chain reaction (touch-down PCR). The PCR products were identified by 1% agarose gel and sequenced in order to further examine the association between Cosmc gene mutation and the susceptibility to HSP.</p><p><b>RESULTS</b>Sequencing results showed two mutations (c.393T>A and c.72A>G) of Cosmc gene in children with HSP. There were no significant differences in the genotype and allele frequencies at the two loci between the HSP and control groups, and this distribution was not associated with sex.</p><p><b>CONCLUSIONS</b>The mutations c.393T>A and c.72A>G in the exon region of Cosmc gene in children with HSP are not associated with the onset of HSP.</p>

Gastric submucosa is inferior to the liver as transplant site for autologous islet transplantation in pancreatectomized diabetic Beagles / 华中科技大学学报（医学）（英德文版）

Intraportal transplantation of islets is no longer considered to be an ideal procedure and finding the extrahepatic alternative site is becoming a subject of high priority. Herein, in this study, we would introduce our initial outcomes of using gastric submucosa (GS) and liver as sites of islet autotransplantation in pancreatectomized diabetic Beagles. Total pancreatectomy was performed in Beagles and then their own islets extracted from the excised pancreas were transplanted into GS (GS group, n=8) or intrahepatic via portal vein (PV group, n=5). Forty-eight hours post transplantation, graft containing tissue harvested from the recipients revealed the presence of insulin-positive cells. All recipients in GS group achieved euglycemia within 1 day, but returned to a diabetic state at 6 to 8 days post-transplantation (mean survival time, 7.16±0.69 days). However, all of the animals kept normoglycemic until 85 to 155 days post-transplantation in PV group (mean survival time, 120±28.58 days; P<0.01 vs. GS group). The results of intravenous glucose tolerance test (IVGTT) confirmed that the marked improvement in glycometabolism was obtained in intrahepatic islet autotransplantation. Thus, our findings indicate that the liver is still superior to the GS as the site of islet transplantation, at least in our islet autotransplant model in pancreatectomized diabetic Beagles.

Berberine Prevents Intestinal Mucosal Barrier Damage During Early Phase of Sepsis in Rat through the Toll-Like Receptors Signaling Pathway

Our previous study has shown berberine prevents damage to the intestinal mucosal barrier during early phase of sepsis in rat through mechanisms independent of the NOD-like receptors signaling pathway. In this study, we explored the regulatory effects of berberine on Toll-like receptors during the intestinal mucosal damaging process in rats. Male Sprague-Dawlay (SD) rats were treated with berberine for 5 d before undergoing cecal ligation and puncture (CLP) to induce polymicrobial sepsis. The expression of Toll-like receptor 2 (TLR 2), TLR 4, TLR 9, the activity of nuclear factor-kappa B (NF-kappaB), the levels of selected cytokines and chemokines, percentage of cell death in intestinal epithelial cells, and mucosal permeability were investigated at 0, 2, 6, 12 and 24 h after CLP. Results showed that the tumor necrosis factor-alpha (TNF-alpha ) and interleukin-6 (IL-6) level were significantly lower in berberine-treated rats compared to the control animals. Conversely, the expression level of tight junction proteins, percentage of cell death in intestinal epithelial cells and the mucosal permeability were significantly higher in berberine-treated rats. The mRNA expression of TLR 2, TLR 4, and TLR 9 were significantly affected by berberine treatment. Our results indicate that pretreatment with berberine attenuates tissue injury and protects the intestinal mucosal barrier in early phase of sepsis and this may possibly have been mediated through the TLRs pathway.

The characteristics and clinical signiifcance ofNOTCH1 mutations in childhood T-cell acute lymphoblastic leukemia / 临床儿科杂志

ObjectiveTo clarify the characteristics and clinical signiifcance of the NOTCH1 mutations in childhood T-cell acute lymphoblastic leukemia (T-ALL).MethodsAmplify and sequence the heterodimerization (HD) domain and the pro-line-glutamicacid-serine-threonine (PEST) domain of theNOTCH1 gene in 28 T-ALL children, in order to explore the frequency, position and type of the mutations as well as their reletions with prognosis.ResultsIn 28 children with T-ALL, 15 cases (51.57%) had been identiifed theNOTCH1 mutations, all of which were heterozygous mutations. The lymphoblast counts in peripher-al blood and bone marrow in theNOTCH1 mutant group at admission were signiifcantly higher than in the non-mutant group (P<0.05). The 1-year remission rate in the 28 children with T-ALL was 75% (21/28), including 80% (12/15) in mutant group in which 3 patients relapsed and all of them died (1-year mortality 20%) and 69.20% (9/13) in non-mutant group in which 4 patients relapsed but all survived (1-year mortality 0%).ConclusionsThe children with T-ALL had a high incidence of NOTCH1 mu-tations at various sites. In addition, the patients withNOTCH1 mutations had more severe disease at diagnosis, better short-term prognosis and poor outcome with salvage therapy after relapse.