Objective To provide clues to diagnosis and treatment for diffuse alveolar hemorrhage (DAH)in patients with diffuse connective tissue diseases(CTD).Method To analyze restropeetively the data of clinical features,pulmonary images and bronchoalveolar lavage(BAL)in 32 patients hospitalized in Peking Union Medical College Hospital from April 2004 to June 2007.Results The data from 1O of the 17 (58.8%)patients with microscopic polyangiitis(MPA),19 of the 1267(1.5%)patients with systemic lupus erythematosus(SLE),2 of the 56(3.6%)patients with Wegener's granulomatosis(WG)and 1 of the 570(0.2%)patients with rheumatoid arthritis(RA)were consistent with the diagnosis of DAH.DAH in SLE occurred in younger patients(mean age at the time of diagnosis 27.3±13.1 years)and early in the course of disease(median duration of SLE from onset was 16.7±18.3 months),while these figures in MPA patients with DAH were 50.1±20.7 years and 10.6±18.7 months.At the time of DAH in SLE.the median systemic lupus erythematosus disease activity index(SLEDAI)score was 17.1±6.7 and anti-ds-DNA antibody titer elevated markedly,while the median C3 level was low.The levels of erythrocyte sedimentation rate(ESR)and C-reactive protein(CRP)of patients with DAH in MPA,WG and RA showed marked elevation.The titer of antieutrophil cytoplasmic antibody(ANCA,MPO/PR3)in patients with MPA and WG was highly positive.The main clinical manifestations of DAH were hemoptysis,dyspnea and rapid decrease of hemoglobulin and hematoerit(HCT)in peripheral blood.Most patients presented with difluse alveolar infiltration on chest X-ray and hish resolution CT.DAH could be confirmed by bloody bronchoscopic lavage.20 patients(62.5%)had secondary pulmonary infections at the time of DAH;fungus and combined bacterial infection were most frequently seen.The mortalitv 0f CTD with DAH was 59.4%(19 out of 32).12 patients (63.2%)with SLE,5 patients(50%)with MPA and both of the 2 patients with WG died.12 of the lethal cases(63.2%)died of respiratory failure.Conclusions CTD patients presenting with hemoptysis and dyspnea with rapid decrease of hemoglobulin,and diffuse alveolar infiltration on chest X-ray or high resolution CT should be seriously considered to be suffering from DAH.A bloody BAL may confirm the diagnosis of DAH.DAH in CTD is an acute,serious and frequently life-threatening situation resulting in respiratory hilure and pulmonary infection.It is important for CTD patients with DAH to be diagnosed early and treated vigorously.

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus (HBV)genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA (NC-siRNA)and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β(IFN-β)and hepatitis B surface antigen (HBsAg)in serum were detected by enzyme linked immunosorbent assay (ELISA).The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction (PCR ).The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was (12.37±5 .32)pg/mL in model group,(22.61 ±6.29 )pg/mL in negative control group,(26.40±5 .39)pg/mL in HBV-siRNA group and (68.37± 21 .00 ) pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group (F =23.988 and 46.523,respectively,both P <0.01).Serum level of HBsAg was (2 864.86±907.11 )ng/mL in model group,(2 198.86±456.89 )ng/mL in negative control group,(1 049.71 ± 396.28 )ng/mL in HBV-siRNA group and (640.86±383.08)ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively).Serum level of HBV DNA was (2.54 ×104 ±1 .46 × 104 )copy/mL in model group,(2.22×104 ±2.62×103 )copy/mL in negative control group,(3.59×103 ±2.88×103 )copy/mL in HBV-siRNA group and (2.65 ×103 ±1 .46×103 )copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F =15 .013 and 16.741 ,respectively,both P <0.05 ).All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.Conclusion HBV-3p-siRNA,which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo ,may be a powerful bifunctional antiviral molecule.

Objective To investigate the effect of early cerebrospinal fluid replacement on nuclear factor-κB (NF-κB) level and clinical outcomes in patients with aneurismal subarachnoid hemorrhage (aSAH) after endovascular embolization.Methods Patients with aSAH received aneurysm embolization were enrolled.They were divided into a cerebrospinal fluid replacement group and a non-cerebrospinal fluid replacement group according to the treatment scheme.All patients were treated with cerebral aneurysm coil embolization within 3 days after admission.The cerebrospinal fluid replacement group performed lumbar puncture cerebrospinal fluid replacement within 24 h after coil embolization,once every other day,20-30 ml of cerebrospinal fluid was replaced each time and 3 mg dexamethasone was injected intrathecally.The NF-κB levels in cerebrospinal fluid were detected at day 1,7 and 14 after the coil embolization.The primary outcome measures were the clinical outcomes determined by the modified Rankin scale (mRS) and the Glasgow outcome scale (GOS) at 3 months after onset.Good outcome was defined as mRS score 0-2 or GOS ＞ 3.The secondary outcome measures included severe complications (hydrocephalus,cerebral vasospasm,cerebral infarction,and rebleeding) and death.Results A total of 81 patients with aSAH received aneurysm embolization were enrolled,including 42 in the cerebrospinal fluid replacement group and 39 in the non-cerebrospinal fluid replacement group.There was no significant differences in the baseline data between the cerebrospinal fluid replacement group and the non-cerebrospinal fluid replacement group (all P ＞0.05).The duration of neck stiffness in the cerebrospinal fluid replacement group was significantly shorter than that in the non-cerebrospinal fluid replacement group (11.3 ± 3.2 d vs.16.5 ± 3.5 d;t =6.985,P ＜ 0.001).The cerebrospinal fluid NF-κB levels were progressively reduced at day 1,7 and 14 after coil embolization in the cerebrospinal fluid rephcement group and non-cerebrospinal fluid rephcement group (all P ＜0.05),but the ccerebrospinal fluid levels of NF-κB in the cerebrospinal fluid replacement group at each time point were significantly lower than those in the non-cerebrospinal fluid replacement group (all P ＜ 0.01).The good outcome rates evaluated according to the mRS score (92.9％ vs.56.4％;x2 =14.446,P ＜ 0.001) and GOS score (97.6％ vs.76.9％;x2 =8.004,P=0.005) in the cerebrospinal fluid replacement group at 3 months were significantly higher than those in the non-cerebrospinal fluid replacement group,and the incidence of cerebral vasospasm was significantly lower than that in the non-cerebrospinal fluid replacement group (14.3％ vs.33.3％;x2 =4.086,P =0.043).Conelusiom Cerebrospinal fluid replacement therapy can reduce the incidence of cerebral vasospasm in patients with aSAH receiving aneurysm embolization and improve clinical outcomes.Its mechanism may be associated with the decrease of NF-κB level in cerebrospinal fluid.

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

ObjectiveTo investigate the association of liver stiffness measurement (LSM) and serum biochemical parameters with hepatic steatosis, liver inflammation, and liver fibrosis in patients with nonalcoholic steatohepatitis (NASH). MethodsA total of 520 patients with NASH who were treated in The Fifth Medical Center of Chinese PLA General Hospital from January 2007 to December 2018 were enrolled, and according to body mass index (BMI) with a cut-off value of 28 kg/m2, the patients were divided into obese group with 151 patients and non-obese group with 369 patients. All patients underwent liver biopsy, and LSM was measured within 3 days before biopsy. Serum biochemical parameters and general clinical data were collected before liver biopsy, and the noninvasive indices aspartate aminotransferase to platelet ratio index (APRI) and fibrosis-4 (FIB-4) were calculated. The t-test was used for comparison of normally distributed continuous between groups, the nonparametric Mann-Whitney U test was used for comparison of non-normally distributed continuous between groups; the chi-square test was used for comparison of categorical data between groups. A Spearman rank correlation analysis was also performed. ResultsAlanine aminotransferase (ALT), aspartate aminotransferase, LSM, controlled attenuation parameter (CAP), spleen length, and APRI gradually increased with the increase in BMI (all P＜0.05). The Spearman correlation analysis showed that LSM, ALT, BMI, and CAP were positively correlated with the degree of hepatic steatosis (r=0.263, 0.327, 0.184, and 0.452, all P＜0.05); LSM, ALT, and CAP were positively correlated with the degree of liver inflammation (r=0.357, 0.278, and 0.121, all P＜0.05); LSM, ALT, BMI, and CAP were positively correlated with the degree of liver fibrosis (r=0.500, 0.216, 0.248, and 0.101, all P＜0.05); age was negatively correlated with the degree of hepatic steatosis, liver inflammation, and liver fibrosis (r=-0.344, -0.129, and -0.163, all P＜0.05). ConclusionLSM, CAP, ALT, and age are significantly correlated with the degree of liver inflammation, liver fibrosis, and hepatic steatosis in NASH patients, and therefore, they can be used in noninvasive diagnostic and predictive models to access the severity of liver injury.

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 .

OBJECTIVETo investigate reliability of FibroScan (FS) in diagnosing size of oesophageal varices (OV) in patients with liver cirrhosis.

METHODSA total of 260 patients with liver cirrhosis were enrolled in the study. All patients underwent endoscopy to assess OV stage and FS to measure liver stiffness. Receiver operating characteristic (ROC) curves were generated to evaluate the diagnostic reliability of FS.

RESULTSThe FS values according to OV stage were 20.9 ± 10.3 kPa for patients without OV, 32.2 ± 13.5 kPa for patients with mild OV, 45.6 ± 18.3 kPa for patients with moderate OV, and 55.1 ± 15.6 kPa for patients with severe OV. Significant differences were found among the groups (P < 0.001) as well as between any two groups (t=6.574, 10.533, 13.247, 4.719, 7.072 and 2.171, P less than 0.05 respectively). ROC curves of FS for the diagnoses of patients with various OV stages showed the following:with vs. without OV, 0.824 (95% CI:77.5% to 87.4%); less than moderate vs. more than moderate OV, 0.849 (95% CI:79.6% to 90.2%); and less than severe vs. severe OV, 0.871 (95% CI:81.1% to 93.0%); the corresponding FS cut-off values were 22.8 kPa, 30.6 kPa, and 34.6 kPa.

CONCLUSIONLiver stiffness measurement by FibroScan allows prediction of the oesophageal varices stage in patients with liver cirrhosis.

Adult ; Elasticity Imaging Techniques ; Esophageal and Gastric Varices ; diagnosis ; etiology ; Female ; Humans ; Liver Cirrhosis ; complications ; diagnosis ; Male ; Middle Aged

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 summarize the clinical features of leukoencephalopathy with vanishing white matter disease (VWM) in children. Methods: A retrospective cohort study was performed on 54 genetically diagnosed VWM patients in Peking University First Hospital from January 2007 to March 2019. Paper registration form and electronic medical record system were used to collect the data,and the children were divided into five groups according to the age of disease onset:<1 year, 1-<2 years, 2-<4 years, 4-<8 years and 8-<18 years respectively. The progression of motor function, episodic aggravation, epileptic seizures, survival time, brain magnetic resonance imaging (MRI) and genotype features were analyzed and compared. Non-parametric test, χ² test or Fisher′s exact test were used for comparison among groups; Kaplan-Meier survival curve was adopted to delineate the survival status of the children. Results: Fifty-four VWM patients were included in the study, including 34 males and 20 females.The age of disease onset was 2 years and 8 months (ranged from 6 months to 9 years and 7 months). Onset age was less than 1 year in 5 cases; onset age was 1-<2 years in 12 cases; onset age was 2-<4 years in 25 cases; onset age was 4-<8 years, in 10 cases; onset age was 8-<18 years in 2 cases; 94% (51/54) of patients had complaint of motor regression at the first visit; 87% (47/54) of patients suffered from episodic aggravation. Episodic seizures occurred in 43% (23/54) patients. In survivors with disease durations of 1-3 years, in 38% (9/24) patients the disease was classified as grades Ⅳ-Ⅴ by gross motor function classification system (GMFCS). For the onset age 1-<2 years group, 1 patient was classified as GMFCS Ⅳ among 3 survivors with disease durations of 1-3 years. As for the 2-<4 years group, 6 patients were classified as GMFCS Ⅳ-Ⅴ among 15 patients with disease durations of 1-3 years, whereas 1 patient was classified as GMFCS Ⅳ-Ⅴ among 4 patients with disease durations of 1-3 years in the 4-<8 years group. Lesions, liquefaction and diffusion restriction in brain MRI were compared among different groups, and it was revealed that the earlier the age of disease onset was, the more likely the subcortical white matter (frontal lobe P<0.01,temporal and parieto-occipital lobe both P=0.002), internal capsule (anterior limb P<0.01, posterior limb P=0.00) and brain stem (midbrain P=0.001, pons P<0.01) were to be involved. In addition, internal capsule (anterior limb P=0.002, posterior limb P=0.005) and brain stem (midbrain P=0.001, pons P=0.003) showed more diffuse restricted diffusion. Moreover, the subcortical white matter (frontal and parieto-occipital lobe both P<0.01, temporal lobe P=0.005) showed earlier rarefaction. The 1-year and 2-year survival rates of the overall patients were 81% and 75% respectively, while the 15-year survival rate was 45%. EIF2B5 gene variation was the most common, which accounts for 43% (23/54), followed by EIF2B3 (22%, 12/54). Conclusions The majority of VWM patients complained of motor regression at the first visit, episodic aggravation and epileptic seizures are common in the course. Earlier age at onset is associated with more rapid clinical progression, shorter survival time as well as more extensive lesions, liquefaction and diffusion restriction in brain MRI. The most common variant gene is EIF2B5, followed by EIF2B3.

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