Objective To investigate the relationship between the serum markers and genotype with viral load in chronic hepati-tis B in order to improve the level of clinical treatment.Methods 819 patients with chronic hepatitis B in our hospital from April 2010 to April 2013were selected as the study subjects and performed the genotyping.The hepatitis B virus(HBV)DNA level was detected by using real-time fluorescence PCR.Results Among the 819 cases of positive HBsAg,357 cases were positive HBV-DNA,accounting for 43.59%;HBeAg(+)accounted for 90.11%;in the positive modes of serum markers and viral load,most of positive cases were HBsAg,HBeAg,HBcAb and HBsAg,HBeAb,HBcAb,accounting for 88.52%;HBV genotyping had B,C,BC mixed types and unclassified type,the logrithm levels of viral DNA were(7.187 ±1.668),(7.184 ±1.558),(7.208 ±1.447)and (7.313±1.505),there was no statistically significant difference among them(P >0.05).Conclusion The virus gene in the patients with chronic hepatitis B is dominated by type C,in which serum markers HBeAg and HBV-DNA viral load have the lose correlated, the infected virus genotype has the low correlation with the viral load.

Limonin existed in citrus fruits has been shown to have anti-bacterial, anti-viral, anti-feedant, anti-nociceptive, anti-inflammatory activities and anti-carcinogenic activities. But the clinical use is limited by its low bioavailability. The aim of this study is to observe the absorption and secretion transport mechanisms of limonin in intestine which can pave the way for the further study and clinical use. The transport characteristics and mechanisms of limonin in rat were studied by in situ intestine perfusion and in vitro Caco-2 cells method. The intestinal absorption of limonin was probably via a facilitated diffusion pathway which was poor and without segment-selection. Verapamil and ketoconazole improved the absorption remarkably according to the result of in vitro Caco-2 cells study; however, probenecid had no significant effect on the absorption. The P-gp efflux and CYP3A4 metabolism were involved in the poor intestinal absorption and low bioavailability of limonin. The exploration of the intestinal absorption mechanism is crucial to the design of dosage form and clinical use of limonin.

The levels of macrophage migration inhibitory factor (MIF) in peripheral blood mononuclear cell (PBMC) and serum in patients with Hashimoto's thyroiditis (HT) were detected by real-time PCR and ELISA,respectively.The results revealed that the expression of MIF mRNA in PBMC ( Z =-4.276,P＜0.01 ) and protein level in serum ( Z=-5.358,P＜0.01 ) were increased in HT patients,and positively correlated with thyroid specific autoantibodies and TSH levels.

Objective To identify the host-cell death pathways (apoptosis, autophagy or necrosis) in L929 cells at the time point of 48 hours post infection (h.p.i.) with Chlamydia muridarum.Methods L929 cells were infected with Chlamydia muridarum at a multiplicity of infection (MOI) of 0.85 for 48 hours.Nuclear fragmentation was observed under fluorescence microscopy following staining L929 cells with DAPI (4′,6-diamidino-2-phenylindole).L929 cells were stained with propidium iodide (PI) plus Annexin Ⅴ and then analyzed by fluorescence-activated cell sorting (FACS) to clarify whether apoptosis or necrosis occurred after Chlamydia muridarum infection.L929 cells were transiently transfected with GFP-LC3 and observed under fluorescent microscopy to analyze cell autophagy.Western blot assay was performed to detect LC3 protein for further analysis of autophagy.Results Apoptosis was not induced in L929 cells by Chlamydia muridarum infection at 48 h.p.i.as no significant nuclear fragmentation was observed.Results of FACS showed that most cells died due to necrosis.Moreover, fluorescent dots of GFP-LC3 formed after infecting transfected L929 cells with Chlamydia muridarum.An increased ratio of LC3Ⅰ to LC3Ⅱ in the L929 cells infected with Chlamydia muridarum was detected by Western blot assay, indicating that autophagy occurred during Chlamydia muridarum infection.Conclusion Necrosis and autophagy rather than apoptosis are induced in L929 cells 48 hours after infection with Chlamydia muridarum.

Objective To clarify the MUM1/IRF4 expression in follicular lymphoma (FL) and its clinical and pathological significance. Methods Ninety-six cases FL were immunostained with MUM1,CD10,bcl-2,bcl-6 and Ki-67 antibodies. The results were compared with their clinical and pathological features. Results The overall MUM1 expression rate in FL was 59.2 % (58/96),including 36.2 % (19/51) grade 1 or 2 and 86.4 %(39/45) grade 3 cases (x2 =24.406,P <0.001). 68.9 % cases with diffuse area were MUM1 positive (x2 =8.161,P =0.004). MUM 1 and CD10 expression had inverse correlation,83.3 % CD10 negative cases were MUM1 positive (x2= 12.649,P<0.001). The mitosis rate and Ki-67 label index were statistically higher in MUM1 positive cases than in negative cases (t = -3.852 & -4.610,respectively,P <0.001). Conclusion MUM1 can be used as a biomarker to divide FL into different malignancies. The MUM1 positive FL may be the feature of high grade non germinal center B cell malignant lymphoma.

Objective To evaluate conventional ultrasound combined with real-time elastography in differential diagnosis of metastatic axillary lymph nodes of breast cancer with a logistic regression model.Methods Conventional ultrasound and real-time elastography were performed in 112 breast cancer patients with 113 axillary lymph nodes.All the cases were confirmed with pathological examinations after surgery. A binary logistic regression model based on ultrasonographic features was developed.A receiver operator characteristic ( ROC) curve was constructed to assess the performance of the model.Results There were 28 nodes with no metastasis and 85 with metastasis in a total of 113 axillary lymph nodes.Three ultrasonographic features including the elastic strain ratio(SR), elastic score and shape were finally selected into the logistic regression model .The correct rate of the logistic regression model for predicting axillary lymph node metastasis was 93.8%, and the area under ROC curve was 0.962.Conclusion The binary classification logistic regression model has a good diagnostic efficacy in the diagnosis of benign and malignant axillary lymph nodes,while real-time elastography can improve the accuracy of conventional ultrasound in the diagnosis of axillary lymph nodes.

OBJECTIVETo study the changes in activity of phosphatidylinositol 4 kinase (PI 4 kinase), phosphatidylinositol 4 phosphate 5 kinase (PIP 5 kinase) and protein kinase C (PKC) during myocardial ischemia and elucidate the relationship between phosphatidylinositol signal pathways and prolonged myocardial ischemia.

METHODSIn vivo an ischemic rat model was used. Activity of PI 4 kinase, PIP 5 kinase and PKC were measured at different times in postischemic heart cells using isotope analysis.

RESULTSThe activity of PI kinase, PIP kinase and PKC in the myocardium increased to peak at 1 hour postischemia, with activities 6.1, 3.0 and 4.0 fold over control levels, respectively. Their activities declined to normal levels with time.

CONCLUSIONThe phosphatidylinositol signal pathway is involved in prolonged myocardial ischemia, but its mechanism needs further study.

1-Phosphatidylinositol 4-Kinase ; metabolism ; Animals ; Male ; Myocardial Ischemia ; enzymology ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; Protein Kinase C ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Signal Transduction

Objective: To investigate the impact of ultrasonic assisted rapid processing technique combined with the environment friendly reagent (which can be utilized in fixing,dehydrating and clearing) on processing tumor biopsy specimens and the subsequent target detection. Methods: Postoperative tissue samples of 56 cases of breast cancer, colorectal cancer, lung cancer, stomach cancer, liver mass, bladder mass, uterus mass were obtained at the National Cancer Center, Cancer Hospital, Chinese Academy of Medical Sciences from February to April, 2017. Three specimens ranging in size from 1 to 3 mm were collected from each sample, and were separated into control group (traditional tissue-processing method); experiment group 1 (3.7% neutral buffered formaldehyde fixation, composite environment friendly reagent and ultrasonic assisted rapid processing) and experimental group 2 (composite environment friendly reagent direct fixation, higher temperature and longer time for tissue processing). Two pathologists blinded to the experimental groups scored totally the nuclear, cytoplasmic, and membrane staining of 43 cases of immunohistochemistry (IHC), four HER2 fluorescence in situ hybridization (FISH), 20 extracted DNA quality and four EGFR gene mutation detection in lung adenocarcinoma; the results were compared with the control group. Results: There was no difference in the IHC staining, HER2 FISH, the DNA quality, and EGFR genetic results between experimental group 1 and control group. For experiment group 2, comparing results of IHC staining, HER2 FISH and the quality of DNA, there was no obvious difference from control group and experiment group 1, but might show an increase in the background of IHC staining. The difference between the treatment temperature and time in the experimental group 2 did not affect the results of the gene mutation detection. Conclusions Environment freindly reagent and ultrasonic assisted rapid processing equipment could be used for rapid processing and diagnosis for tumor biopsies. Using complex environment-friendly reagents supplement fixation, higher treatment temperature and longer treatment time do not significantly affect the IHC, FISH and molecular detection accuracy.