A chemical method to prepare biotin-labeled nucleic acid probe for nonisotopic hybridization is introduced. The method involves the bisulfite-catalyzed transamination reaction of cytosine residues in nucleic acids with ethylenediamine or hexanediamine. Primary amino groups on the cytosine derivatives are then reacted with biotiny-N-hydroxysuccinimide ester. Results of dot blot and Southern blot carried out with biotinlabeled ? DNA probe show that such nonisotopic probe method is sensitive, specific, repeatable and unexpensive.

The conditions for denaturation of DNA and RNA were invertigated. The results show that one minute to heat DNA at 100鈩?is sufficient for denaturation of DNA (several hundreds bp to 23kb). DNA degraded more obviously with longer neating time and the degradation decreased when DNA was dissdved in TE buffer. But no difference of degree of RNA degradation was found in there three conditions for RNA denaturation.

Objective To analyze the clinical and imaging characteristics of congenital sensorineural hearing loss (CSNHL)children combined with white matter (WM)lesions in order to provide evidence for clinical practice. Methods With referral to the Department of Pediatrics,Peking University First Hospital from November 201 1 to De-cember 201 5,documents of 78 patients of CSNHL combined with WMlesions were collected and analyzed for the clini-cal and imaging characteristics.Results Bilateral severe -profound hearing loss existed in all 78 cases,48.1 %(25 /52 cases)of the patients exhibited gross motor development delay,98.1 %(51 /52 cases)of them had normal cognition development.One hundred percent (61 /61 cases)of patients had abnormal language development.Infection occurring during pregnancy existed in 21 .2%(1 1 /52 cases)of the patients,the premature and smaller for the gestational age in-fants accounted for 28.9% (1 5 /52 cases).The bilateral multiple WMlesions from the brain MRI were in dot to flake sizes with sharp boundary,the intensity of T1 -weighted imaging decreased,T2 -weighted imaging and fluid attenuated inversion recovery increased.Eighty -two point one percent (64 /78 cases)of the patients were found to have the periventricular and subcortical WM involvement.The most frequently affected periventricular region was the posterior horn (91 .9%,68 /74 cases),followed by the anterior horn and temporal horn,and the least with the body involvement. The former three had a combined lesion tendency (55.4% -68.9%).There was an extensive involvement in the sub-cortical WMof parietal,frontal,temporal and occipital lobes respectively(73.5% -88.2%).Subcortical WM involve-ment of multiple lobes was common (accounted for 67.6% -77.9%).The enlargement of bilateral ventricles existed in 37.2%(29 /78 cases)of the patients and cystic changes in the subcortical WM of anterior temporal lobe could be found in 9.0% (7 /78 cases)patients.Calcification in 2 CT cases was reported.Corpus callosum and basal ganglia of all cases were normal.For cases with MRI scans more than once,WMlesions of 96.0%(24 /25 cases)patients became silent or self -restored.Conclusions The clinical presentations of CSNHL combined with WM lesions are mild,not paralleled with their multiple foci.It is considered as demyelination or a delay of myelination.Due to its benign course, it is probably not the contraindication for the cochlear implantation.

SARS coronavirus (SARS-CoV) develops an antagonistic mechanism by which to evade the antiviral activities of interferon (IFN). Previous studies suggested that SARS-CoV papain-like protease (PLpro) inhibits activation of the IRF3 pathway, which would normally elicit a robust IFN response, but the mechanism(s) used by SARS PLpro to inhibit activation of the IRF3 pathway is not fully known. In this study, we uncovered a novel mechanism that may explain how SARS PLpro efficiently inhibits activation of the IRF3 pathway. We found that expression of the membrane-anchored PLpro domain (PLpro-TM) from SARS-CoV inhibits STING/TBK1/IKKε-mediated activation of type I IFNs and disrupts the phosphorylation and dimerization of IRF3, which are activated by STING and TBK1. Meanwhile, we showed that PLpro-TM physically interacts with TRAF3, TBK1, IKKε, STING, and IRF3, the key components that assemble the STING-TRAF3-TBK1 complex for activation of IFN expression. However, the interaction between the components in STING-TRAF3-TBK1 complex is disrupted by PLpro-TM. Furthermore, SARS PLpro-TM reduces the levels of ubiquitinated forms of RIG-I, STING, TRAF3, TBK1, and IRF3 in the STING-TRAF3-TBK1 complex. These results collectively point to a new mechanism used by SARS-CoV through which PLpro negatively regulates IRF3 activation by interaction with STING-TRAF3-TBK1 complex, yielding a SARS-CoV countermeasure against host innate immunity.
Dimerization ; HEK293 Cells ; Humans ; I-kappa B Kinase ; metabolism ; Interferon Regulatory Factor-3 ; metabolism ; Interferon Type I ; antagonists & inhibitors ; metabolism ; Membrane Proteins ; chemistry ; genetics ; metabolism ; Papain ; metabolism ; Peptide Hydrolases ; chemistry ; metabolism ; Phosphorylation ; Protein Binding ; Protein Structure, Tertiary ; Protein-Serine-Threonine Kinases ; metabolism ; SARS Virus ; enzymology ; Signal Transduction ; TNF Receptor-Associated Factor 3 ; metabolism ; Ubiquitination

Spontaneous bacterial peritonitis (SBP) is a common complication of cirrhosis. Based on our current understanding of SBP, the most common etiologies for SBP in cirrhosis are Enterobacter and Streptococcal species. Th e Aeromonas species are ubiquitous in fresh or sea water. Aeromonas caviae is never identified as etiology in cases of SBP. A patient, who had a history of liver cirrhosis related to chronic hepatitis B virus infection for 1 year, presented with diarrhea. He had diarrhea 1 week later returned from coastal city. He was hospitalized and treated with norfloxacin after 7 days of severe symptoms, including fever, abdominal distention, and diarrhea. Analysis of the ascitic specimen revealed a white-cell count of 4.42 × 109 cells/L with 88% neutrophils. Analysis of stool specimen showed a white-cell count of 60 cells per high-power field. Th e patient started the injection of cefriaxone at a dose of 4 g/d. However, the situation was not improved. Th ree days later, stool and ascitic fluid culture showed positive for Aeromonas caviae. Antibiotic susceptibility testing revealed that imipenem, meropenem, amikacin, and cefoperazone-sulbactam were highly sensitive to the Aeromonas caviae. However, the bacilli resisted to ceftriaxone, ceftazidime, ampicillin-sulbactam, levofloxacin, and sulfamethoxazole. Ceftriaxone was then switched to imipenem. The patient was fully recovered 14 days later. Aeromonas caviae is a rare pathogen of SBP in cirrhosis. It resists to third-generation of cephalosporin and fluroquinolone, which are of frequently used dependent on clinical experience. It needs a special attention.
Aeromonas caviae ; Anti-Infective Agents ; Ascitic Fluid ; Gram-Negative Bacterial Infections ; pathology ; Humans ; Leukocyte Count ; Liver Cirrhosis ; Male ; Microbial Sensitivity Tests ; Peritonitis ; microbiology ; pathology

Objective To evaluate the effectiveness and safety of direct-acting antiviral agents (DAA) treatment in Chinese chronic hepatitis C (CHC) patients with genotype (GT) 1b HCV infection in a real world setting .Methods The consecutive GT1b CHC Chinese patients treated with sofosbuvir (SOF) plus daclatasvir (DCV) (n＝62) or SOF plus ledipasvir (LDV) (n＝171) were enrolled from July 2014 to December 2016 at 302 Military Hospital of China .The treatment duration for all the patients was 12 weeks .All the clinical parameters were measured at baseline and then 4-weekly till 12 weeks after the end-of-treatment (EOT ).Baseline clinical characteristics ,treatment efficacy ,safety and tolerance were compared .Serum HCV RNA concentration was detected by means of COBAS TaqMan assay with a lower detection limit of 15 IU/mL ,and liver stiffness was measured using FibroScan?.Sustained virologic response (SVR) was defined as HCV RNA under the lower limit of quantification 12 weeks after EOT (SVR12).Students′t-test ,pearson χ2 test ,Spearman rank correlation analysis and Fisher exact test were used for comparison between groups when appropriate .Results Among 233 patients ,173 cases had baseline HCV RNA level ≥ 6 .0 lg IU/mL and 97 cases hade liver stiffness measurement (LSM )≥17.5 kPa.The baseline liver inflamation ,liver fibrosis ,and HCV RNA load of patients in the two groups were not significantly different (all P＞0 .05).The HCV RNA of all the 233 patients was undetectable at the end of 12-week treatment ,while 2 patients relapsed after 12 weeks of EOT with the overall SVR12 of 99.1% .HCV RNA decline was significantly faster in patients with lower LSM than those with higher LSM (ρ＝0 .233 ,P＝0 .001) ,and SVR12 was higher in those with lower LSM .In terms of other clinical characteristics of SOF+DCV and SOF+LVD groups ,alanine transaminase declined from (68 .0 ± 60 .1) and (70 .1 ± 56 .1) U/L to (21 .1 ± 10 .9) U/L and (15 .3 ± 9 .5) U/L ,respectively ,total bilirubin declined from (21 .3 ± 17 .3) and (18 .2 ± 14 .0) μmol/L to (13 .2 ± 6 .7) and (10 .2 ± 4 .6) μmol/L , respectively ,AFP declined from 19 .6 (10 .6 ,62 .3) and 15 .0 (12 .0 ,25 .0) μg/L to 6 .5(4 .5 ,18 .7) and 7 .8(5 .3 ,15 .4) μg/L ,respectively ,LSM declined from 17 .6 (8 .9 ,25 .4) and 15 .7 (7 .8 ,23 .9) kPa to 13.9(6 .5 ,21 .4) and 9 .1(5 .6 ,19 .9) kPa ,respectively ,serum album elevated form (37 .5 ± 5 .8) and (38 .7 ± 5 .5) g/L to (41 .3 ± 4 .7) and (42 .8 ± 5 .1) g/L ,respectively ,platelet elevated from (120.9 ± 78 . 2)×109/L and (136 .6 ± 65 .8 )× 109/L to (139 .5 ± 71.8 )× 109/L and (149 .7 ± 71.4 )× 109/L , respectively .Reports of adverse events were low in both groups .Conclusions Both SOF + DCV and SOF/LDV therapy are highly effective with ＞ 98% of SVR12 and reduce LSM value significantly with good safety for CHC GT1b Chinese patients .

Objective:To evaluate the treatment effect of hemoperfusion(HP) on Henoch-Sch?nlein purpura(HSP) outcomes.Methods:PubMed, Cochrane library, Web of science, Wanfang database, CNKI and CBM database were searched from inception to February 2020.Literatures of randomized controlled trials(RCTs) that investigated the effect of HP on HSP outcomes were included.Articles screening, data extraction and quality assessment were accomplished by two investigators independently, and statistical analyses were performed by RevMan 5.3.Results:Thirteen RCTs were included with 803 cases, of which, 397 cases were in the HP group while 406 cases were in the control group.The Meta-analysis revealed the HP group had less disappearing time of hematuresis or albuminuria[ MD=-2.77, 95% CI(-3.18, -2.36), P<0.001], relieving time of abdominal pain[ MD=-1.70, 95% CI(-2.17, -1.23), P<0.001], disappearing time of hematochezia[ MD=-1.54, 95% CI(-1.68, -1.40), P<0.001], and the length of hospital stay[ MD=-3.23, 95% CI(-3.60, -2.87), P<0.001] than the control group. Conclusion:The HP could shorten disappearing time of hematuresis or albuminuria, relieving time of abdominal pain, disappearing time of hematochezia, and the length of hospital stay of HSP.

Autophagy plays important roles in modulating viral replication and antiviral immune response. Coronavirus infection is associated with the autophagic process, however, little is known about the mechanisms of autophagy induction and its contribution to coronavirus regulation of host innate responses. Here, we show that the membrane-associated papain-like protease PLP2 (PLP2-TM) of coronaviruses acts as a novel autophagy-inducing protein. Intriguingly, PLP2-TM induces incomplete autophagy process by increasing the accumulation of autophagosomes but blocking the fusion of autophagosomes with lysosomes. Furthermore, PLP2-TM interacts with the key autophagy regulators, LC3 and Beclin1, and promotes Beclin1 interaction with STING, the key regulator for antiviral IFN signaling. Finally, knockdown of Beclin1 partially reverses PLP2-TM's inhibitory effect on innate immunity which resulting in decreased coronavirus replication. These results suggested that coronavirus papain-like protease induces incomplete autophagy by interacting with Beclin1, which in turn modulates coronavirus replication and antiviral innate immunity.
Apoptosis Regulatory Proteins ; antagonists & inhibitors ; genetics ; immunology ; Autophagy ; Beclin-1 ; Coronavirus NL63, Human ; genetics ; immunology ; Gene Expression Regulation ; HEK293 Cells ; HeLa Cells ; Host-Pathogen Interactions ; immunology ; Humans ; Immune Evasion ; Immunity, Innate ; Interferon-gamma ; genetics ; immunology ; Lysosomes ; metabolism ; virology ; MCF-7 Cells ; Membrane Fusion ; Membrane Proteins ; antagonists & inhibitors ; genetics ; immunology ; Microtubule-Associated Proteins ; genetics ; immunology ; Papain ; genetics ; immunology ; Phagosomes ; metabolism ; virology ; RNA, Small Interfering ; genetics ; immunology ; Signal Transduction ; Virus Replication

Transketolase(TKT)is an important enzyme that catalyzes the non-oxidative phase group transfer reac-tion of pentose phosphate pathway(PPP),and is involved in the metabolism of various energy substances in the body,such as glucose,ribose,nucleotides and lipids.TKT can reduce oxidative stress,inflammation,atheroscle-rosis,endothelial dysfunction and Tau protein phosphorylation by inhibiting advanced glycated end-produces(AGEs)produced by non-enzymatic glycosylation(NEG)of proteins and lipids in a high-glucose environment im-proving blood glucose,glucose tolerance and β cell function so to prevent and treat diabetes and its complications.This article reviews research progress on the mechanism of TKT in the treatment of diabetes mellitus and its compli-cations.

China attaches great importance to the occupational health assistance for small and medium-sized enterprises, but many efforts are still needed. Through the research and comparative analysis of the policies and measures of occupational health assistance for small and medium-sized enterprises at domestic and international, this paper finds that there are still some problems in the occupational health assistance for small and medium-sized enterprises in China, such as imperfect policies, lack of safeguard measures, support platforms and resources, and puts forward that China can promote the occupational health assistance for small and medium-sized enterprises from the aspects of perfecting the support system, strengthening safeguard measures, building relevant platforms and enriching resources.