This study investigated the role of Echinococcus multilocularis vesicle-derived extracellular vesicles(EVs)con-taining emu-miR-10-5p in regulating HSC activation in vitro.Mice were injected intraperitoneally with protoscoleces to con-struct a secondary alveolar echinococcosis model,and E.multilocularis vesicles were cultured in mice and identified by PCR.The EV structure in cultured vesicle tissue was observed by transmission electron microscopy.EVs were isolated from vesicle culture supernatants through differential-ultracentrifugation,and identified by transmission electron microscopy,nanoparticle size analysis,and western blotting.The activity of HSCs treated with different EV concentrations was detected with CCK-8 assays.The expression levels of α-SMA,and collagen Ⅰ and Ⅲ were detected by RT-qPCR and western blotting after EV treatment of HSCs.Bioinformatics analysis identified the specific miRNA emu-miR-10-5p.The proliferation of HSCs was detected with EdU after overexpression of emu-miR-10-5p,and the expression levels of α-SMA,collagen Ⅰ,and collagen Ⅲ in cells were detected by RT-qPCR and western blotting.Detection of α-SMA,collagen Ⅰ,and collagen Ⅲ expression levels in HSCs was performed after EVs were extracted from the culture supernatant of vesicular tissues with emu-miR-10-5p knockdown.E.multilocularis vesicles were successfully cultured from focal material in mice,and the E.multilocularis origin was confirmed through PCR amplification of specific genes.TEM observation of culture vesicles in-dicated abundant surface vesicular structures.E.multilocularis vesicle-derived EVs were isolated by differential-ultracentrifu-gation,and confirmed by TEM to be spherical and membrane like-structures.Vesicle-derived EVs were internalized by HSCs,and promoted the proliferation and activation of HSCs.Comprehensive analysis of miRNA sequencing results revealed emu-miR-10-5p expression in E.multilocularis-infected mouse serum,metacestode tissue,E.multilocularis-derived EVs,germi-nal layer cells,and protoscoleces.Moreover emu-miR-10-5p was confirmed to be present in higher amounts in E.multilocu-laris-derived EVs,according to RT-qPCR.Further studies confirmed that overexpression of emu-miR-10-5p promoted HSC proliferation and increased α-SMA,collagen Ⅰ,and collagen Ⅲ mRNA and protein expression levels in HSCs.In contrast,EVs extracted from culture supernatants after emu-miR-10-5p knockdown in vesicles did not increase a-SMA,collagen Ⅰ,and colla-gen Ⅲ mRNA and protein expression levels.E.multilocularis vesicle-derived EVs and emu-miR-10-5p thus promote the acti-vation of HSCs.

Little is known about the non-structural protein 6(NSP6)of rotavirus.This report describes expression of the NSP6 of a group A human rotavirus strain TB-Chen in bacteria,and its immunological properties and cellular distribution.The results showed that the recombinant NSP6(rNSP6)was expressed in high efficiency without any other proteins fused(possesses about 34.2% of total bacterial proteins).rNSP6 elicited mono-specific antibodies in immunized guinea pigs and the antibodies could react with the rNSP6 itself and the viral NSP6 proteins synthesized in SA11-or Wa-infected MA104 cells in Western blot and immunofluorescence assay.The NSP6 distributed evenly in the cytoplasm mainly around the nucleus of virus-infected cells,no viroplasm-like gathering observed;The top amount of NSP6 synthesized in SA11-infected cells or Wa-infected cells could be detected at 12h after infection.This is the first report about the high expression of entire NSP6(without any other proteins fused)in prokaryotic expression system and detection of NSP6 synthesis in virus infected cells by immunofluorescence assay.The results are important to understand the structure,biological properties and further application of the NSP6.

By means of genetic cloning and recombinant techniques, full genome cDNA sequences of rotavirus strain TB-Chen were isolated from an infantile hospitalized with acute gastroenteritis. Nucleotide sequences analyses showed that the full genome of strain TB-Chen contains 18613 nucleotides, encoding 5791 amino acids. Genotyping results showed that the strain TB-Chen belongs to genotype G2P[4]/NSp4[A]. This is the first report on a full genome of Group A rotavirus in China, and has important significance for deep understanding structure and functions of rotaviruses and developing rotavirus vaccines.

OBJECT: To obtain values of normal human lung and diffuse pathological changes, in order to provide a simple and convenient diagnostic method for measuring human visceral organs in the autopsy of pathology and forensic pathology. METHODS: The exact mass, volume and density of normal and pathological lung were synchronously measured with the intelligentized volume-densimeter made by authors. RESULTS: The date-base about the volume and density of human lung with different races were established for anatomy, anthropology and biodynamics. In two cases of over-load in circulation and water toxicosis, the severe lung edema was proved with the scale of lung density, which was atypical edema in the lung tissue. CONCLUSION Measurement of visceral organs in the autopsy can assist to an integrative pathological diagnosis.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Anthropometry/methods* ; Autopsy ; Child ; Child, Preschool ; Female ; Forensic Medicine ; Humans ; Infant ; Infant, Newborn ; Lung/pathology* ; Male ; Middle Aged ; Organ Size ; Pulmonary Edema/pathology* ; Sex Factors ; Young Adult

OBJECTIVETo investigate the association of serum vitamin D [25-(OH)D] level with the severity and treatment in children with Henoch-Schönlein purpura (HSP).

METHODSA total of 50 children with newly-diagnosed HSP between January and December, 2015 were enrolled as HSP group, and 49 healthy children were enrolled as control group. Fasting serum samples were collected, and ELISA was used to measure serum 25-(OH)Dlevel. According to the serum 25-(OH)Dlevel, the HSP group were further divided into normal group (>20 ng/mL) (n=9), insufficiency group (15-20 ng/mL) (n=15), deficiency group (≤15 ng/mL) (n=25), and severe deficiency group (≤5 ng/mL) (n=1). The general data, clinical manifestations, hormone therapy, course of disease before admission, and length of hospital stay were compared between groups.

RESULTSThe HSP group had a significantly lower serum 25-(OH)Dlevel than the control group (16±6 ng/mL vs 29±5 ng/mL; P<0.01). Compared with the normal and insufficiency groups, the deficiency and severe deficiency groups had significant increases in the incidence rate of renal involvement, rate of hormone application, and median length of hospital stay (P<0.05), while there was no significant difference in course of disease before admission (P>0.05).

CONCLUSIONSChildren with HSP have a low serum 25-(OH)Dlevel, and such children may have a high risk of renal involvement, a high rate of hormone application, and a prolonged length of hospital stay. However, further studies are needed to investigate whether vitamin D supplementation is helpful to the treatment of HSP and can shorten the course of disease in children with HSP.

Child ; Female ; Humans ; Length of Stay ; Male ; Purpura, Schoenlein-Henoch ; blood ; complications ; drug therapy ; Severity of Illness Index ; Vitamin D ; analogs & derivatives ; blood

Objective To study the prevalence of esophageal cancer and various lesions of esophagus in high risk areas through a screening program for early diagnosis and treatment.Methods Random cluster sampling method was used to select some portions of a natural village as screening object in the high risk areas of esophageal cancer,from 2006 to 2011.Endoscope iodine staining and index biopsy screening methods were used on people with high risk and followed by pathological exams for confirmation.Results The detection rates regarding mild esophageal hyperplasia,moderate and severe esophageal hyperplasia were 5.33％ (803/15 065),1.28％( 193/15 065 ),0.68％( 102/15 065 ) respectively while the detection rates on carcinoma in situ,intramucosal carcinoma and invasive cancer were 0.15％(22/15 065),0.06％(9/15 065),0.29％(43/15 065)respectively.The detection rate in male esophageal hyperplasia was higher than in female.People younger than 65 years old,the detection rates on mild,moderate or severe esophageal hyperplasia and invasive cancer showed an increase with age,with the 60- year-olds group reaching the highest.The detection rates on the above said diseases were 7.72％( 198/2565 ),2.07％(53/2565),1.29％( 33/2565 ),0.51％ ( 13/2565 ) respectively.The detection rates on mild,moderate or severe esophageal hyperplasia varied in different years and with statistically significant differences (P＜0.001) but did not show any obvious trend of changing.Geographical distribution of mild esophageal hyperplasia,moderate esophageal hyperplasia,severe esophageal hyperplasia also significantly varied in different villages (P＜0.001).The highest detection rate in the mountainous villages was seen the highest while the detection rate of village from hilly areas was the lowest.Conclusion There were considerable numbers of patients with precancerous lesions in the general population from the high risk areas.The detection rate of esophageal cancer in the mountain residents was higher than the rate in the hilly areas.Men and the elderly were the key populations calling for esophageal cancer prevention programs to be carried out.

Poly (ADP-ribose) polymerase-1 (PARP-1) plays as a double edged sword in cerebral ischemia-reperfusion, hinging on its effect on the intracellular energy storage and injury severity, and the prognosis has relationship with intervention timing. During ischemia injury, apoptosis and oncosis are the two main cell death pathway sin the ischemic core. The participation of astrocytes in ischemia-reperfusion induced cell death has triggered more and more attention. Here, we examined the protective effects and intervention timing of the PARP-1 inhibitor PJ34, by using a mixed oxygen-glucose deprivation/reperfusion (OGDR) model of primary rat astrocytes in vitro, which could mimic the ischemia-reperfusion damage in the "ischemic core". Meanwhile, cell death pathways of various PJ34 treated astrocytes were also investigated. Our results showed that PJ34 incubation (10 μmol/L) did not affect release of lactate dehydrogenase (LDH) from astrocytes and cell viability or survival 1 h after OGDR. Interestingly, after 3 or 5 h OGDR, PJ34 significantly reduced LDH release and percentage of PI-positive cells and increased cell viability, and simultaneously increased the caspase-dependent apoptotic rate. The intervention timing study demonstrated that an earlier and longer PJ34 intervention during reperfusion was associated with more apparent protective effects. In conclusion, earlier and longer PJ34 intervention provides remarkable protective effects for astrocytes in the "ischaemic core" mainly by reducing oncosis of the astrocytes, especially following serious OGDR damage.
Animals ; Apoptosis ; Astrocytes ; cytology ; drug effects ; Cell Survival ; Cells, Cultured ; Glucose ; deficiency ; Humans ; Lactate Dehydrogenases ; metabolism ; Male ; Models, Biological ; Oxygen ; metabolism ; Phenanthrenes ; pharmacology ; Poly(ADP-ribose) Polymerase Inhibitors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects

To develop a fed-batch fementation process of E. coli TOP10 containing a recombinant plasmid pBAD/HBs Fab. Cells were grown in semi-defined medium at 37 degrees C, and the feed operation using glycerol as carbon source was performed when dissolved oxygen increased. When the target cell concentration reached to 64g/L, arabinose was added to a final concentration of 0.02%. Cells were grown for another 5h with the culture temperature decreased from 37 degrees C to 30 degrees C. In the whole process, cell growth was monitored by measuring OD600 of samples taken at 1/2h intervals and the dissolved oxygen was kept above 30%. After the fementation, E. coli pellets were collected for purification of Fab protein. The specificity of Fab protein was confirmed by Western blot, and binding activity to HBsAg was verified by Dot blot. Cell concentration we got is 96g wet bacteria per liter, the Fab protein is about 6% of total protein of the host, that is 80mg per liter. Stable fermentation parameters were obtained for fermentation to improve productivity of the Fab protein. The Fab protein was produced in the form of soluble biologically active protein, it's better than inclusion bodies from which biologically active protein can only be recovered by complicated and costly denaturation and refolding process.
Blotting, Western ; Escherichia coli ; genetics ; metabolism ; Fermentation ; physiology ; Hepatitis B Surface Antigens ; immunology ; Immunoglobulin Fab Fragments ; biosynthesis ; genetics ; immunology ; Temperature

OBJECTIVETo investigate the expression of guanylin in colorectal cancer.

METHODSThe expression of guanylin was examined by RT-PCR and semiquantitative analysis in 20 cases of colorectal cancer, and its relationship with clinical characteristics was analyzed.

RESULTSThe positive expression of guanylin in normal tissue (80%, 16/20) was significantly higher than that in tumor tissue (35%, 7/20) (P<0.01). The same result was found in the semiquantitative analysis of 14 cases with differential expression. Differential expression of guanylin in colorectal cancer was associate with TNM stage (P<0.05), not with sex, Borrmann type and degree of differentiation (all P>0.05).

CONCLUSIONThere is differential expression of guanylin in colorectal cancer, and this kind of differential expression is associated with tumor TNM stage.

Colorectal Neoplasms ; metabolism ; pathology ; Female ; Gastrointestinal Hormones ; metabolism ; Humans ; Male ; Middle Aged ; Natriuretic Peptides ; metabolism ; Neoplasm Staging

BACKGROUNDTourette syndrome (TS) is a complex, heterozygous genetic disorder. The number of molecular genetic studies have investigated several candidate genes, particularly those implicated in the dopamine system. The dopamine D3 receptor (DRD3) gene has been considered as a candidate gene in TS. There was not any report about the association study of TS and DRD3 gene in Han Chinese population. We combined a case-control genetic association analysis and nuclear pedigrees transmission disequilibrium test (TDT) analysis to investigate the association between DRD3 gene rs6280 single nucleotide polymorphisms (SNPs) and TS in a Han Chinese population.

METHODSA total of 160 TS patients was diagnosed by the diagnostic criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. The DRD3 gene rs6280 SNPs were genotyped by TaqMan SNP genotyping assay technique in all subjects. We used a case-control genetic association analysis to compare the difference in genotype and allele frequencies between 160 TS patients and 90 healthy controls. At the same time, we used TDT analysis to identify the DRD3 gene rs6280 transmission disequilibrium among 101 nuclear pedigrees.

RESULTSThe genotype and allele frequency of DRD3 gene rs6280 SNPs had no statistical difference between control group (90) and TS group (160) (χ2 = 3.647, P = 0.161; χ2 = 0.643, P = 0.423) using Chi-squared test. At the basis of the 101 nuclear pedigrees, TDT analysis showed no transmission disequilibrium of DRD3 gene rs6280 SNPs (χ2 = 0; P = 1).

CONCLUSIONSOur findings provide no evidence for an association between DRD3 gene rs6280 and TS in the Han Chinese population.

Adolescent ; Child ; Female ; Gene Frequency ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; genetics ; Receptors, Dopamine D3 ; genetics ; Tourette Syndrome ; etiology