AIM:To explore the regulatory effect of microRNA-3666 (miR-3666) on the expression of its tar-get gene phosphatase and tensin homologue deleted on chromosome ten (PTEN) in leukemic cells.METHODS: miR-3666 expression levels in normal peripheral blood mononuclear cells and leukemic cells were determined by quantitative real-time PCR.miR-3666 targeting PTEN 3'-untranslated region (3'UTR) was predicted by TargetScan software .3'UTR of PTEN was inserted in the dual luciferase reporter vector psiCHECK 2.The reporter activity was evaluated by the Dual-Lu-ciferase Reporter Assay System after the luciferase promoter vector and miRNA were co -transfected into HEK293T cell line. K562 cells were transfected with synthetic miR-3666 inhibitor ( anti-miR-3666) or a synthetic control miRNA ( anti-miR-C) .The expression of PTEN protein in the above transfected K 562 cells was determined by Western blotting .RESULTS:miR-3666 was up-regulated in the human leukemic cell lines and primary leukemic cells compared to normal peripheral blood mononuclear cells .The results of dual luciferase assays validated PTEN as a specific target gene of miR-3666.Inhi-bition of miR-3666 resulted in an up-regulation of PTEN protein expression in the K 562 cells.CONCLUSION:miR-3666 is over-expressed in leukemic cells .The abnormal over-expression of miR-3666 may play a key role in leukemia due to the down-regulation of PTEN .

Objective To observe effects of different concentrations of sevoflurane combined with propofol on recovery quality in patients undergoing partial hepatic resection.Methods Seventy-eight patients,aged 20-70 years old,selected for partial hepatic resection were randomly divided into three groups,26 patients in each group:total intravenous propofol anesthesia group (group T), propofol combined with 0.5 MAC sevoflurane anesthesia group (group S1),propofol combined with 1.0 MAC sevoflurane anesthesia group (group S2).Spontaneous respiration recovery time,recovery time,extubation time and modified Aldrete score of 9 time were recorded after operation.Modified OAA/S scores as well as modified Aldrete score at extubation immediate time (T1 )and 5 min (T2 ), 1 5 min(T3 ),30 min(T4 )after extubation were also recorded.Results Total amount of propofol in groups S1,S2 significantly less than group T and total amount of propofol in group S2 significantly less than group S1(P <0.05).The recovery time,extubation time,modified Aldrete score of 9 time in groups S1 and S2 were significantly shorter than group T (P <0.05).Modified OAA/S scores at T1 ,T2 and the modified Aldrete scores at T1 in both groups S1 and S2 were significantly higher than group T,while group S2 was significantly higher than group S1 (P <0.05).Conclusion Compared with total intravenous propofol anesthesia,both propofol combined with 0.5 MAC sevoflurane and propofol combined with 1.0 MAC sevoflurane anesthesia improves the recovery quality in patients un-dergoing partial hepatic resection,and the recovery time was decreased in propofol combined with 1.0 MAC sevoflurane anesthesia.

Aim To investigate the effects of cholecystokinin-octopeptide on IL-12 secreted in murine bone marrow-derived dendritic cells induced by lipopolysaccharide.Methods The CCK receptor subtypes were investigated by immunofluorescence in murine bone marrow-derived dendritic cells.Enzyme linked immunosorbent assay and Western blot were used to estimate the contents of IL-12 and p38MAPK activity.Results CCK-1R and CCK-2R were detected in BM-DC;CCK-8(at concentrations of 10-10,10-8,10-6 mol?L-1)could significantly increase the secretion of IL-12 in the LPS-induced DC, and LPS-activated p38MAPK activity in a dose-dependent manner.The effect of CCK-8 was reduced partially by CR1409(a CCK-1R antagonist) and CR2945(a CCK-2R antagonist).Conclusion CCK-8 could dose-dependently increase the expressions of IL-12 in LPS-induced DC,probably by promoting p38MAPK activity and the effect was mediated by CCK1 and CCK2 receptors.

BACKGROUND:Establishing a characteristic.stable and repeatable model of Th1/Th2 imbalance in animals,is the key of studying the mechanism of Th1/Th2 imbalance.OBJECTIVE:To observe the characteristics of Th1/Th2 imbalance in splenocytes derived from Balb/C mice immnnized by keyhole limpet hemocyanin(KLH).DESIGN:A randomized control exploratory experiment.SETTING:Hebei Provincial Forensic Laboratory.Institute of Basic Medicine,Hebei Medical University.MATERLALS:The experiment was carried out in the Hebei Provincial Forensic Laboratory,Institute of Basic Medicine,Hebei Medical University from September 2005 to January 2007.Balb/C mice were adopted in this study.and all the disposals were in accordance with the guidance of animal ethics.METHODS:Balb/C mice were immunized with KLH emulsified in complete Freund's adjuvant(CFA),splenocytes were acquired,and the peak of cytokine secretion was determined in 3 groups:KLH groups of 6.25 mg,kg.12.5 mg,kg and 25 mg/kg.According to the immunizing dose and immunizing frequency.mice were divided into 7 groups:KLH groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,secondary immunity groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,as well as control group.According to the determined levels of IgG1 and IgG2a in blood serum.mice were divided into KLH group of 6.25 mg/kg and control group.MAIN OUTCOME MEASURES:Mice splenocytes supematant was detected with enzyme linked immunosorbent assay (ELISA)for the production of Th1 cytokines interferon (IFN)-γ,interleukin(IL)-2.IL-12 p40 and Th2 cytokines IL-4 and IL-5.The levels of Th1 antibody IgG2a and Th2 antibody IgG1 in blood serum were also detected by ELISA.RESULTS:The spleen derived from KLH-immunized mice appeared hypertrophy,and the number of splenocytes was manifold.Splenocytes restimulated with KLH in vitro produced much more IFN-γ,IL-2,IL-4,IL-5,but not IL-12p40.IL-2 secretion was obviously elevated after incubated for 24 hours and achieved pinnacle at 48 hours;productions of IL-4,IL-5 and IFN-γ were elevated after 24 hours,and increased gradually to 96 hours;IL-12p40 production was very low at every time point.Using different doses of KLH inlmunity once or twice,could all lead to the elevated productions of IL-2,IL-4,IL-5,IFN-γ,and the elevation of IL-4/IFN-γ ratio,but the secondary immunity group of 6.25 mg/kg KLH showed obviously higher levels than other groups(P＜0.01).The level of KLH specific antibody IgG2a and IgG1,especially IgG1 was elevated in blood serum of KLH-immunized mice.CONCLUSION:Balb/C mice immunizad with KLH emulsified in CFA can indce a Th2 predominant imbalance in splenocytes.

AIM: To study the effects of cholecystokinin octapeptide (CCK-8) on T-lymphocyte subsets in keyhole limpet haemocyanin (KLH)-immunized mice.METHODS: Female BALB/c mice were immunized with KLH and injected with different dosages of CCK-8 or saline simultaneously. Positive CD4+and CD8+ T cells in peripheral blood or splenocytes were measured by flow cytometry. The production and mRNA level of Th1 cytokine, IFN-? and Th2 cytokine, IL-4 in the splenocytes were evaluated by ELISA and RT-PCR, respectively. In addition, lung tissue sections were made with HE staining.RESULTS: CCK-8 downregulated the positive CD4+and CD8+ T lymphocytes both in peripheral blood and in splenocytes in KLH-immunized mice, resulting in the reduction of CD4+/CD8+ ratio.CCK-8 improved the production of IFN-?, elevated IFN-? gene transcription, whereas cut down the production of IL-4, decreased IL-4 gene transcription. CCK-8 lightened the pulmonary inflammation induced by KLH.CONCLUSION: CCK-8 inhibits CD4+T lymphocytes activation, Th2 cytokine mRNA expression and protein production in KLH-immunized mice, indicating that CCK-8 modulates adaptive immune response.

Objective:To investigate the changes in structural and functional connectivity of different hippocampal subregions in courses of Alzheimer's disease (AD).Methods:One hundred and seventeen subjects from Sino Longitudinal Study on Cognitive Decline (SILCODE) were chosen; their cognitive functions were assessed using neuropsychological tests, and brain beta-amyloid (Aβ) levels were measured using PET-MRI. These patients were divided into group of normal cognitive function and negative Aβ ( n=68), group of normal cognitive function and positive Aβ ( n=29), and group of cognitive impairment and positive Aβ ( n=20); neuropsychological characteristics and changes in volume, structural connectivity and functional connectivity of hippocampal subareas were compared among the 3 groups. Results:Compared with group of normal cognitive function and negative Aβ and group of normal cognitive function and positive Aβ, group of cognitive impairment and positive Aβ had significantly decreased scores of Mini-mental State Examination (MMSE), Montreal Cognitive Assessment-Basic Version (MoCA-B), Auditory Verbal Learning Test (AVLT)-Short-term Delayed Recall (SR), AVLT-Long-term Delayed Recall (LR), Animal Fluency Test (AFT) and Boston Naming Test (BNT), and statistically increased Shape Trails Test B (STT-B) scores ( P<0.05). Compared with group of normal cognitive function and negative Aβ and group of normal cognitive function and positive Aβ, group of cognitive impairment and positive Aβ had significantly decreased volumes of the left and right hippocampal CA1, CA2, CA3, dentate gyrus (DG), entolorhinal cortex (EC) and inferior support area (Sub, P<0.05). Compared with group of normal cognitive function and negative Aβ, group of normal cognitive function and positive Aβ had increased or decreased structural connectivity, and almost all increased functional connectivity of each hippocampal subregion; compared with group of normal cognitive function and negative Aβ, group of cognitive impairment and positive Aβ had almost all increased structural connectivity, and almost all decreased functional connectivity of each hippocampal subregion. Conclusion:Changes in gray matter volume and structural and functional connectivity in various hippocampal subregions are different in courses of AD.

OBJECTIVE: To detect potential variant in an ethical Han Chinese pedigree affected with breast cancer. METHODS: The proband and her relatives were subjected to next-generation sequencing using a target capture sequencing kit containing 121 cancer-related genes. Candidate variants were selected by analysis of their type, frequency in population, and segregation with the phenotype. Candidate variant was verified by Sanger sequencing and TA cloning. RESULTS: A c.2013_2014ins GT variant was detected in the BRCA1 gene among all breast cancer patients from this pedigree but not among healthy females. The variant was not recorded in the 1000 Genome Project database or the Exome Aggregation Consortium (ExAC) database. The frameshifting insertion was predicted to form an premature stop codon in gene transcript and can give rise to a truncated protein. CONCLUSION The BRCA1 c.2013_2014ins GT variant probably underlies the pathogenesis of breast cancer in this Chinese pedigree.
Asian Continental Ancestry Group ; BRCA1 Protein ; genetics ; Breast Neoplasms ; genetics ; Exome ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Pedigree ; Phenotype

Objective:To explore a fast and accurate method to diagnose children's pneumonia according to respiratory signals, so as to avoid the cancer induction caused by traditional X-ray examination.Methods:A Mach Zehnder optical fiber sensor was used to build a respiratory signals(RSPs) detection system, and the RSPs of the monitored children were extracted according to the vibration signal generated by the children's lung rales. Preprocessing methods such as the discrete cosine transform(DCT) were used to compress and denoise the RSPs. Multi-feature extraction of RSPs was conducted through signal processing methods such as the Hilbert transform and autoregressive (AR) model spectrum estimation. A support vector machine (SVM) classification model was constructed to classify the collected RSPs.Results:The accuracy rate of the proposed RSP classification of children with or without pneumonia was 94.41%, which was higher than the previous methods.Conclusions:The children's pneumonia diagnosis system based on an optical fiber sensor has a higher detection accuracy, and is expected to be widely used in clinical practice.

Objective To investigate whether elevated parathyroid hormone (PTH) levels could induce endothelial - to - mesenchymal transition (EndMT) and adipocyte transition in endothelial cells (ECs), and to determine the possible underlying mechanism. Methods (1) A rat model of secondary hyperparathyroidism and chronic kidney disease (CKD) was established. The adiposity in bone marrow was detected by oil red O staining. Immunofluorescence staining was performed to detect the expression and localization of cluster of differentiation 31 (CD31) and fibroblast-specific protein 1 (FSP1). (2) The human umbilical vein ECs were cultured in vitro. Western blotting was performed to detect protein expressions of EndMT-related markers CD31, FSP1 and α-smooth muscle actin (α-SMA) in interference groups with different PTH concentrations (0, 10-11, 10-9, 10-7 mol/L PTH for 48 h) and times (0, 12, 24, 48 h, 10-7 mol/L PTH), as well as the expression of β-catenin in interference groups with different PTH concentrations. The localizations of CD31, FSP1 and β - catenin were observed by cell immunofluorescence. Protein expressions of adipocytes markers peroxisome proliferator - activated receptor-γ (PPAR-γ) and CCAAT/enhancer binding protein-α (C/EBP-α) by Western blotting and the degree of adipogenesis by oil red O staining were detected after transformed ECs were cultured in adipogenic culture medium for one week. Small interfering RNA (siRNA) was performed to silenceβ - catenin expression. ECs were divided into control siRNA group, β - catenin siRNA group, PTH +control siRNA group and PTH+β-catenin siRNA group. Protein expressions of CD31, FSP1 and PPAR-γby Western blotting and the degree of adipogenesis by oil red O staining were determined. Results (1) In vivo, compared with the control, CKD rats had increased adipocytes in bone marrow (P＜0.05), and the co-expression of CD31 and FSP1 in bone marrow ECs. (2) In vitro, PTH significantly inhibited the expression of endothelial marker CD31 and increased the expressions of mesenchymal markers FSP1 and α-SMA in concentration-and time-dependent manners. These indexes in 10-7 mol/L PTH group and 0 mol/L PTH group, in 48 h group and 0 h group showed statistical differences (all P＜0.05). In PTH group ECs with 10-7 mol/L PTH for 48 h showed FSP1 accumulation in the cytoplasm and reduced expressions of CD31, and ECs had higher expressions of PPAR-γ and C/EBP-α as well as the degree of adipogenesis than those in control group (all P＜0.05). Furthermore, PTH enhanced the nuclearβ-catenin protein levels in ECs in concentration-dependent. The expressions of β-catenin in 10-7 mol/L PTH group and 0 mol/L PTH group showed statistical differences (P＜0.05). β - catenin expressed in the cytoplasm in control group, while it enter into the nucleus in PTH group. Compared with those in PTH+control siRNA group, the expressions of CD31 and PPAR-γ as well as the degree of adipogenesis decreased in PTH+β-catenin siRNA group (all P＜0.05), while the expression of FSP1 increased (P＜0.05). Conclusions PTH induces ECs - to - adipocytes transition by the canonical Wnt/β - catenin signaling pathway, which might account for bone loss in CKD. Silenced β - catenin expression can inhibit PTH-induced EndMT and adipogenesis.