Objective To provide clues for the study on the mechanism of autoimmune liver disease (AILD) by exploring the existence of specific bacteria in liver tissues of AILD patients.Methods From August 2017 to August 2018,at Department of Gastroenterology arnd Hepatology,Tianjin Medical University General Hospital,a total of 12 patients diagnosed as AILD (four autoimmune hepatitis (AIH),four primary biliary cirrhosis (PBC) and four PBC-AIH overlap syndrome (OS)) and four patients with hepatic cyst (control group) were enrolled and all the patients underwent liver biopsy.16S rRNA gene sequencing was carried out in the obtained aseptic liver tissues.Linear discriminant analysis effect size was used to find out the specific bacteria.Spearman correlation analysis was performed to analyze the correlation between the liver microbiota and the disease.The metabolic function of the 16S rRNA gene sequences was also predicted.Results Bacteria were detected in the liver tissues of all the 16 patients.At the species level,the abundance of Planococcus rifietoensis of AIH group was 0.100％,which was higher than those of other three groups (0),and the difference was statistically significant (linear discriminant analysis (LDA) =3.31,P =0.034).The abundance of Anoxybacillus flavithermus of PBC group was 0.200％,which was higher than those of other three groups (0.100％),and the difference was statistically significant (LDA =3.34,P =0.014).The abundance of Pseudomonas aeruginosa PAO1,Bacillus firmus,Brevibacillus agri,Acinetobacter baumannii,Sphingomonas zeae and Salmonella enterica were significantly negatively correlated with serum level of γ-glutamyl transferase (r=-0.68,-0.68,-0.67,-0.68,-0.68 and-0.66,all P ＜0.01).Compared with that of the hepatic cyst group,the lipid metabolism of AILD patients decreased.The levels of serum low density lipoprotein and total cholesterol were significantly negatively correlated with the biosynthesis of unsaturated fatty acids (r =-0.55 and-0.65,both P ＜ 0.05).Conclusions There exist specific bacteria in the liver tissues of AIH and PBC groups.The liver microbiota which is closely related with the pathogenesis of AILD might be a potential therapeutic target and diagnostic biomarker.

BACKGROUNDThe fat derived protein adiponectin plays an important role in the regulation of glucose metabolism. The aim of this study was to provide the experimental basis for further investigating on adiponectin (ADPN) function. Its eukaryotic recombinant was constructed and expressed in precursor cells of 3T3-L1 adipocytes. The effects of dexamethasone on peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA expression in 3T3-L1 cells with human recombinant adiponectin were assessed.

METHODSThe recombinant plasmid pMD18-T-hADPN and eukaryotic expression vector pcDNA3.1(+) were digested by two restrictive endonucleases and adiponectin and linear pcDNA3.1(+) were obtained. Then, they were ligated and translated into JM109. The recombinant pcDNA3.1(+)-hADPN so obtained was identified by digestion by restrictive endonuclease and nucleotide sequencing. The 3T3-L1 precursor cells were transfected using SuperFect Transfection Reagent (Qiagen). Furthermore, 3T3-L1 cells with human recombinant adiponectin incubated with dexamethasone (0.5 mmol/L) for 24 hours, cells were collected and total RNA was extracted. The PPAR-gamma mRNA expression was quantified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSAfter eukaryotic recombinant was digested by Hind III and EcoR I, fragments of 800 bp and 5.4 kb were identified by nucleotide sequence scanning and consistent with theoretical values. Electrophoretogram of RT-PCR in 3T3-L1 precursors showed only one band in front of 250 bp, which was consistent with theoretical value 234 bp. In the 3T3-L1 cells, 3T3-L1 cells with plasmid and 3T3-L1 cells human recombinant adiponectin, treatment with dexamethasone (0.5 mmol/L) decreased PPAR-gamma mRNA expression compared to untreated controls (P < 0.01). Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

CONCLUSIONThe 3T3-L1 cells stably transfected human recombinant adiponectin had increased PPAR-gamma mRNA expression. Dexamethasone suppressed PPAR-gamma mRNA expression in the 3T3-L1 cells. Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

3T3-L1 Cells ; Adiponectin ; physiology ; Animals ; Dexamethasone ; pharmacology ; Insulin Resistance ; Mice ; PPAR gamma ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

This study aims to investigate the function of two SNPs (rs8904C > T and rs696G >A) in 3' untranslated region (3'UTR) of NFKBIA gene by constructing luciferase reporter gene. A patient's genomic DNA with rs8904 CC and rs696 GA genotype was used as the PCR template. Full-length 3'UTR of NFKBIA gene was amplified by different primers. After sequencing validation, these fragments were inserted to the luciferase reporter vector, pGL3-promoter to construct recombinant plasmids containing four kinds of haplotypes, pGL3-rs8904C/rs696G, pGL3-rs8904C/rs696A, pGL3-rs8904T/rs696G and pGL3-rs8904T/rs696A. Then these plasmids were transfected into LS174T cells and the luciferase activity was detected. Compared with pGL3-vector transfected cells (negative control), the luciferase activity of the four kinds of recombinant plasmids was significantly decreased (P < 0.001). For rs696G > A, the luciferase activity of the recombinant plasmids containing A allele (pGL3-rs8904C/rs696A and pGL3-rs8904T/rs696A) was about 45.1% (P < 0.05) and 56.1% (P < 0.001) lower than those containing G allele (pGL3-rs8904C/rs696G and pGL3-rs8904T/rs696G), respectively. For rs8904C > T, there were no significant differences in the luciferase activity between the recombinant plasmids containing T allele and those with C allele. Together, the luciferase reporter gene vectors containing SNPs in NFKBIA gene 3'UTR were constructed successfully and rs696G > A could decrease the luciferase activity while rs8904C >T didn't have much effect on the luciferase activity.
3' Untranslated Regions ; Genes, Reporter ; Genetic Vectors ; Humans ; I-kappa B Proteins ; genetics ; Luciferases ; NF-KappaB Inhibitor alpha ; Plasmids ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Transfection

OBJECTIVETo objectively assess the clinical efficacy and safety of a new Pulian Ointment (, NPLO) in treating psoriasis of blood-heat syndrome of Chinese medicine.

METHODSA total of 108 patients with psoriasis of blood-heat syndrome were equally assigned, using a randomizing digital table, to the test group treated externally with NPLO and the control group treated with placebo; the medication was done using a singleblinded method twice a day. Meanwhile, all patients received by oral intake a conventional Chinese decoction for clearing heat and cooling blood; the therapeutic course was 4 weeks for both groups. The therapeutic efficacy, changes in the Psoriasis Area and Severity Index (PASI) score and various aspects of the lesion, including scaly eruption, erythema, infiltration, size, score of itching as well as adverse reactions were observed.

RESULTSThe trial was completed in 100 patients, 51 in the test group and 49 in the control group. The remarkably effective rate was 45.10% and the total effective rate was 84.31% in the test group, which were significantly higher than those in the control group, 12.24% and 51.02%, respectively, showing a significant difference between groups (P<0.01). The test group also showed better effects in the improvement of the PASI score of the lesions and scores on erythema, infiltration, size of lesion as well as itching. No adverse event was found in either group.

CONCLUSIONNPLO is a Chinese remedy for the external treatment of psoriasis of the blood-heat syndrome with a reliable therapeutic efficacy and good safety.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Ointments ; Psoriasis ; drug therapy ; Syndrome

Objective To investigate the clinical effect of external ventricular drainage on the prognosis of anterior communicating artery aneurysms.Methods Retrospectively collected and analyzed 96 patients of anterior communicating artery aneurysms who were treated in our hospital from June 2013 to October 2015,and they were divided into the observation group which was given external ventricular drainage treatment and the control group which was not given external ventricular drainage treatment.These patients were followed up for 6 months to 2 years,and the results of the 2 groups were graded according to the analysis of postoperative complications and the Glasgow prognostic score (GOS).Meanwhile,evaluated the general function of the patients according to the KPS score.Results The the incidence rate of complications after treatment in observation group was 54.17％,which was lower than 86.96％ in the control group,and the difference was statistically significant(P ＜ 0.05).The cure rate of observation group was 79.16％,which was higher than 50％ in the control group,and the difference was statistically significant (P ＜ 0.05).The postoperative KPS score in the observation group was (79.68 ± 13.24) points,which was higher than (62.57 ± 12.72) points in the control group,and the differences were statistically significant (P ＜ 0.05).Conclusion External ventricular drainage can reduce the compression injury of the brain tissue to a minimum degree,reduce intracranial pressure,relieve cerebral edema caused by intracranial pressure,reduce complications,and improve the prognosis of patients and the cure rate.

Objective To investigate the clinical efficacy of small bone-flap decompression of posterior fossa through neuroendoscopy in the treatment of ChiariⅠmalformation.Methods Retrospectively analyzed the clinical data of 44 cases of ChiariⅠdeformity/syringomyelia who were admitted into affiliated hospital of Xuzhou medical university from January 2013 to December 2016.And divided these patients into the observation group ( n=22,treated with small bone-flap decompression of posterior fossa through neuroendoscopy ) and the control group ( n=22,treated with posterior fossa decompression/tonsillectomy) according to surgical methods .The recovery time and the postoperative com-plications of the two groups were analyzed ,and the long-term efficacy and changes in spinal cord cavity were observed through the follow-up of 1 to 20 months.Results There was no death case in both of the two groups .The effective rate of the two groups were 86.36％and 86.36％respectively.The complication rate of the observation group was 27.35％, which was significantly lower than 77.3％of the control group with statistical significance(P<0.05).There was no significant differences in the recovery time ,postoperative curative effect and spinal cord cavi-ty changes between the two groups (P>0.05).Conclusion The treatment of Chiari Ⅰmalformation with small bone-flap decompression of posterior fossa through neuroendoscopy is a new surgical procedure with less trauma ,shorter hospital stay ,less complication and less effective compared with traditional method .

AIMTo observe the expressional alterations of colony stimulating factor-1 receptor (CSF-1R) after ischemic injury of cerebral cortex, and study the function of colony stimulating factor-1 (CSF-1)/CSF-1R signal during the process of ischemic injury and repair of central nervous system (CNS).

METHODSWe examined the distribution and expression of CSF-1R in normal brain tissues and ischemic brain tissues by immunohistology and Western blot analysis.

RESULTSThe expression of CSF-1R in neurons could be up-regulated by ischemic injury in CNS.

CONCLUSIONCSF-1/CSF-1R might take part in the process of ischemic injury and repair.

Animals ; Brain Ischemia ; pathology ; physiopathology ; Cerebral Cortex ; blood supply ; Female ; Macrophage Colony-Stimulating Factor ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; Neurons ; metabolism ; Random Allocation ; Receptor, Macrophage Colony-Stimulating Factor ; genetics ; metabolism ; physiology ; Reperfusion Injury ; metabolism ; physiopathology

BACKGROUNDThe mechanism of cerebral vasospasm following subarachnoid haemorrhage (SAH) is not understood. Here, we hypothesized that apoptosis of endothelial cells induced by p53 and its target gene em dash p53 upregulated modulator of apoptosis (PUMA) played an important role in development of cerebral vasospasm. We also observed the effects of a p53 inhibitor, pifithrin-alpha (PFT-alpha), on reducing the expression of p53 and PUMA, consequently decreasing the apoptosis of endothelial cells and alleviating cerebral vasospasm.

METHODSMale Sprague-Dawley rats weighing 300-350 g were randomly divided into five groups: a control group (sham surgery), a SAH group, a SAH+dimethyl sulfoxide (DMSO) group, a SAH + PFT-alpha (0.2 mg/kg) group and a SAH + PFT-alpha (2.0 mg/kg) group. PFT-alpha was injected intraperitoneally immediately after SAH. Rats were sacrificed 24 hours after SAH. Western blot and immunohistochemical staining were used to detect the levels of p53, PUMA and caspase-3 protein. In addition, mortality and neurological scores were assessed for each group. Statistical significance was assured by analysis of variance performed in one way ANOVA followed by the Tukey test. The neurological and mortality scores were analyzed by Dunn's method and Fisher exact test, respectively.

RESULTSAfter SAH, Western blot and immunohistochemical staining showed the levels of p53, PUMA and caspase-3 in the endothelial cells and the numbers of TdT mediated dUTP nick end labelling (TUNEL) positive endothelial cells were all significantly increased in the basilar arteries (P<0.05), but significantly reduced by PFT-alpha (P<0.05). These changes were accompanied by increasing diameters and declining wall thickness of basilar arteries (P<0.05), as well as reduced mortality and neurological deficits of the rats (P<0.05).

CONCLUSIONSPFT-alpha could protect cerebral vessels from development of vasospasm and improve neurological outcome as well as reduce the mortality via suppressing apoptosis induced by p53 in the endothelial cells of cerebral vessels.

Animals ; Apoptosis ; drug effects ; Benzothiazoles ; pharmacology ; therapeutic use ; Blotting, Western ; Disease Models, Animal ; Endothelial Cells ; drug effects ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Subarachnoid Hemorrhage ; complications ; drug therapy ; pathology ; physiopathology ; Toluene ; analogs & derivatives ; pharmacology ; therapeutic use ; Tumor Suppressor Protein p53 ; physiology ; Vasospasm, Intracranial ; prevention & control

Purpose To investigate the clinicopathologic features and differential diagnosis of oncocytic variant of medullary thyroid carcinoma (MTC).Methods Morphological,immunohistochemical findings on 3 oncocytic variant of MTC cases and electron microscopic findings on 2 of these 3 cases were studied,with review of the relevant literatures.Results The cytoplasm was abundant,eosinophilic and granular with defined margins.The nuclei were enlarged,round to oval.Prominent nucleoli were identified in some cells.Tumor cells formed sheets,trabeculae or follicles with infiltrative pattern.Immunohistochemically,thyroglobulin (TG) was negative in all 3 cases.Calcitonin was positive in 2 cases and negative in 1 case.Ultrastructurally,a large number of mitochondria and various neuroendocrine granules were found in the 2nd and 3rd cases.Conclusion Oncocytic variant of MTC is very rare with variable histopathologic appearances.It should be considered in diagnosing oncocytic lesions of thyroid.A definite diagnosis can be rendered based on comprehensive findings of the immunohistochemistry,serology study and electron microscopy.It needs to avoid misdiagnosis and missed diagnosis.

OBJECTIVETo evaluate antiviral effects of Peg-IFNa-2a in patients with chronic hepatitis B.

METHODS92 chronic hepatitis B patients were enrolled to receive the treatment with Peg-IFNa-2a 180 μg subcutaneous injection once weekly. The patients who did not get early response were divided into 3 groups: group 1, extend the treatment to 72 weeks; group 2, combined with nucleus(s)ide analogue (entecavir or adefovir) treatment; group 3, continue the treatment until 48 weeks. HBV DNA and quantitative HBsAg were assessed at baseline, week 12, 24, 36 and after 24 weeks follow-up.

RESULTSPatients in group 1 had significantly higher SVR rate (78.3%) than patients in group 3 (38.1%, X2=7.33, P<0.05). The mean reduction of HBsAg in group 1 at 24 weeks of post-treatment follow up was higher than that in group 3 (t=2.11, P<0.05). In group 2 the mean reductions of HBV DNA at 24 weeks of post-treatment follow up were (3.9+/-1.1) log10 copy/ml and (3.7+/-1.3) log10 copy/ml respectively with combination of entecavir or adefovir, both of which were significantly higher than that in group 3(t=8.45 and 6.31, P<0.05); the SVR rates in the entecavir group and the adefovir group (83.3% and 85.7%, respectively) were significantly higher than that in group 3 (X2=8.20 and 7.78, P<0.05); the mean reductions of HBsAg in the entecavir group and the adefovir group [(0.8+/-0.5) log10 IU/ml and (0.9+/-0.3) log10 IU/ml, respectively ] were significantly greater than group 3[(0.4+/-0.3) log10 IU/ml, t=3.05 and 4.58, P<0.05]. The level of HBV DNA and C genotype were the main predictors of response.

CONCLUSIONIndividualizing therapy by prolonging the duration of Peg-IFNa-2a treatment to 72 weeks or adding nucleoside analogues such as entecavir and adefovir in patients without early response may substantially increase the SVR rate and lead to the decrease of HBsAg.

Adenine ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; therapeutic use ; Female ; Guanine ; analogs & derivatives ; therapeutic use ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Organophosphonates ; therapeutic use ; Treatment Outcome