The Metabolic Reprogramming Profiles in the Liver Fibrosis of Mice Infected with Schistosoma japonicum.

Disordered glucose and lipid metabolism contributes to the progression of several liver diseases, while the upregulation of phosphatase and tensin homology deleted on chromosome ten (PTEN), a well-known tumour suppressor gene, can improve the condition through metabolic programming. This study first characterized the metabolic profiles and the involvement of PTEN in the hepatic fibrosis induced by Schistosoma japonicum (S. japonicum) to provide a novel clue for metabolism-targeted treatment. Compared with control mice, infected mice showed infiltrated immune cells in their livers, increased levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and decreased glucose levels in their sera. The expression of key enzymes in the glycolytic pathway was significantly increased, and the expression of gluconeogenic genes was distinctly decreased. Moreover, the infection upregulated the hepatic expression of enzymes involved in fatty acid oxidation, which was consistent with the decreased number of lipid droplets in livers and the lowered levels of triglyceride in sera. Consistently, PTEN and its downstream signalling were significantly inhibited. In vitro, soluble egg antigen (SEA) downregulated the expression of PTEN in both the macrophage RAW264.7 cell line and the murine hepatocellular carcinoma HEP1-6 cell line, and induced a metabolic phenotype similar to the in vivo results. Overall, this study showed that S. japonicum infection induced the reprogramming of glucose and lipid metabolism in mice during the period of liver fibrosis and that SEA could act as a modulator to trigger such a metabolic switch in macrophages and hepatocytes. PTEN might play an essential role in mediating these metabolic reprogramming events.

[Delayed DNA double-strand break in S-phase H1299 cells after thermal damage].

OBJECTIVE: To study the pattern of DNA double-strand break (DSB) formation in S-phase cells after thermal damage and explore the mechanisms behind heat sensitivity of S-phase cells and delayed DSBs. METHODS: Flow cytometry was used to analyze the cell cycle arrest in H1299 cells exposed to thermal damage, and EdU incorporation assay was employed to evaluate the DNA replication capacity of the cells. The cells synchronized in S phase were obtained by serum starvation and DSBs were observed dynamically using neutral comet assay. Trypan blue dye exclusion technique was used to analyze the cell viability after thermal damage. Western blotting (WB) was used to detect the phosphorylation of ATM and DNA binding RAD18. RESULTS: The percentage of S-phase cells increased significantly after exposure of the cells to 45 degrees celsius; for 1 h (P<0.01). The time-dependent variation pattern of EdU incorporation was similar to that of S-phase cell fraction. The comet tail began to appear only after incubation of the cells at 37 degrees celsius; for some time and the Olive tail moment (OTM) increased with prolonged incubation. Cell death remained low until 7.5 h after heat exposure of the S-phase cells and then increased rapidly. The phosphorylation of ATM first increased but then decreased drastically. In cells with heat exposure, DNA binding RAD18 was attenuated obviously compared that in non-exposed cells. CONCLUSION: Thermal damage causes cell cycle arrest in S phase, and delayed fatal DSBs occur in the arrested cells due to persistent replication and DNA damage repair suppression, which are the possible cause of heat sensitivity of S-phase cells.

[Testing of germination rate of hybrid rice seeds based on near-infrared reflectance spectroscopy].

Germination rate of rice seeds was measured according to technical stipulation of germination testing for agricultural crop seeds at present. There existed many faults for this technical stipulation such as long experimental period, more costing and higher professional requirement. A rapid and non-invasive method was put forward to measure the germination rate of hybrid rice seeds based on near-infrared reflectance spectroscopy. Two varieties of hybrid rice seeds were aged artificially at temperature 45 degrees C and humidity 100% condition for 0, 24, 48, 72, 96, 120 and 144 h. Spectral data of 280 samples for 2 varieties of hybrid rice seeds with different aging time were acquired individually by near-infrared spectra analyzer. Spectral data of 280 samples for 2 varieties of hybrid rice seeds were randomly divided into calibration set (168 samples) and prediction set (112 samples). Gormination rate of rice seed with different aging time was tested. Regression model was established by using partial least squares (PLS). The effect of the different spectral bands on the accuracy of models was analyzed and the effect of the different spectral preprocessing methods on the accuracy of models was also compared. Optimal model was achieved under the whole bands and by using standardization and orthogonal signal correction (OSC) preprocessing algorithms with CM2000 software for spectral data of 2 varieties of hybrid rice seeds, the coefficient of determination of the calibration set (Rc) and that of the prediction set (Rp) were 0.965 and 0.931 individually, standard error of calibration set (SEC) and that of prediction set (SEP) were 1.929 and 2.899 respectively. Relative error between tested value and predicted value for prediction set of rice seeds is below 4.2%. The experimental results show that it is feasible that rice germination rate is detected rapidly and nondestructively by using the near-infrared spectroscopy analysis technology.

[Preparation of anti-BRDT-NY polyclonal antibody and expression of BRDT-NY protein in digestive tract tumors].

AIM: To prepare the mouse polyclonal antibody against human BRDT-NY prokaryotic protein and analyze the expression of BRDT-NY protein in digestive tract tumors. METHODS: The N-terminal amino acids of BRDT-NY protein was amplified by PCR and cloned into the expression vector pET28a(+). The recombinant plasmid pET28a+-BRDT-NY was transformed into E.coli BL21 and induced to express the recombinant protein with IPTG. We immunized BALB/c mice with the purified BRDT-NY protein for preparing the specific polyclonal antibody. The titer of the antibody was analyzed by ELISA. The expression of BRDT-NY protein in digestive tract tumors was detected by immunohistochemistry. RESULTS: Recombinant human BRDT-NY protein was expressed in BL21 and purified successfully by Ni-affinity chromatography. Two months after the mice were immunized with the purified BRDT-NY protein, we obtained the specific polyclonal antibody of high titer 1:100 000. Immunohistochemical analysis revealed that BRDT-NY protein was highly expressed in digestive tract tumors. CONCLUSION: The successful preparation of mouse polyclonal antibody against BRDT-NY lays a foundation for the research on the role of BRDT-NY protein in the pathology of human digestive tract tumors.

[The clinical value of balloon dilatation through flexible bronchoscope in the management of tracheobronchial stenosis in 149 cases of endobronchial tuberculosis].

OBJECTIVE: To explore the clinical value of balloon dilatation through flexible bronchoscope in the management of tracheobronchial stenosis of endobronchial tuberculosis. METHODS: From January 2005 to September 2009, 149 cases of tracheobronchial stenosis caused by endobronchial tuberculosis were examined by flexible bronchoscope and treated with balloon dilatation. Changes of the clinical features, atelectasis and airway diameters were observed and evaluated before and after the last treatment and in 12 months. RESULTS: The airway diameters were immediately enlarged (100%, 149/149) after the procedure, and the clinical symptoms were relieved. The average airway diameter changed from (2.7 ± 1.4) mm before the procedure, to (6.8 ± 2.0) mm, (6.4 ± 1.7) mm and (6.3 ± 2.3) mm immediately, 3 and 12 months after the treatments. Expansion of atelectasis was seen in 92% (34/37) of the cases, and the rate of restenosis was 3.4% (5/146) 12 months after treatment. There were significant differences before and after the treatments in the airway diameters, expansion rate of atelectasis and the general outcome (t = 13.09-20.50, P < 0.01), but there were no differences among measurements immediately, 3 and 12 months after the treatments. The final effective rate was 93.3% (139/149). Severe complications (4.0%, 6/149) were rare in these patients. CONCLUSION: Balloon dilatation through flexible bronchoscope is a simple, effective and safe method for the management of tracheobronchial stenosis after endobronchial tuberculosis.

[Efficacy of cryoablation combined with CpG oligonucleotides in the treatment of murine transplanted colon carcinoma].

OBJECTIVE: To investigate the efficacy of cryoablation combined with CpG ODN in the treatment of murine transplanted colon carcinoma. METHODS: Colon carcinoma model was established by subcutaneously inoculating CT26 cells into the right flank in BALB/c mice. The tumor-bearing mice were randomly divided into 4 groups:the group of PBS injected in peritumoral area, the group of cryoablation, the group of cryoablation combined with CpG ODN, the group of CpG ODN injected in peritumoral area. The tumor size changes were measured. Serum levels of interleukin (IL)-12 and interferon-gamma(IFN-gamma) were assayed by ELISA. The rates of CD3(+)CD4(+)T, CD3(+)CD8(+)T lymphocytes in serum were counted with flow cytometry. Mice in the cryoablation group and the combined group with tumor regression were re-challenged with CT26 cells. RESULTS: The survival time of cryoablation group and combined therapy group were (80.3 + or - 5.4) days and (83.8 + or - 5.5) days, respectively, longer than (53.7 + or - 3.7) days in PBS group and (51.5 + or - 6.8) days in CpG ODN group(all P<0.05). The suppress rates of tumor cells in cryoablation group and combined therapy group were 83.8% and 86.2% respectively. After 20 days following treatment, CD3(+)CD4(+)T/CD3(+)CD8(+)T ratio and the concentrations of IL-12 and IFN-gamma in mice serum of cryoablation group and combined therapy group were higher than those in PBS group and CpG ODN group(all P<0.05). No significant difference was found in CD3(+)CD4(+)T/CD3(+)CD8(+)T ratio between cryoablation group and combined therapy group(P>0.05). However, the concentrations of IL-12 and IFN-gamma in combined therapy group were higher than those of cryoablation group(all P<0.05). After re-challenging, tumor formation rate in the cryoablation combined with CpG ODN group was 16.7%, significantly lower than that in the cryoablation group(83.8%)(P<0.05). CONCLUSION: Cryoablation combined with CpG ODN can increase antitumor immune response in mice, and therefore can decrease the tumor formation when re-challenged with CT26 cells.

[Therapeutic and immunological effects of microwave ablation combined with CpG ODN on transplanted colon carcinoma in mice].

OBJECTIVE: To investigate the therapeutic and immunological effects of microwave ablation (MA) combined with CpG ODN in mice bearing transplanted colon carcinoma. METHODS: A mouse model bearing colon carcinoma was established by subcutaneously inoculating CT26 cells into the right flank of Balb/c mice. The tumor-bearing mice were randomized into control group with PBS injection in the peritumoral area, MA group, MA combinated with CpG ODN group, and CpG ODN group with CpG ODN injection in the peritumoral area. The tumor volume changes were observed, and serum CD3(+)CD4(+) and CD3(+)CD8(+) T lmyphocytes were analyzed by flow cytometry after the treatments. Serum levels of interleukin (IL)-2, IL-12 and IFN-gamma were detected by ELISA. The mice in the MA group and the combined treatment group showing tumor regression were rechallenged with CT26 cells. RESULTS: No significant difference was found in the number of serum CD3(+), CD3(+)CD4(+), or CD3(+)CD8(+) T lymphocytes between the 4 groups. The ratio of CD3(+)CD4(+)/CD3(+)CD8(+) T lymphocytes in the combined treatment group and MA group were 1.58-/+0.10 and 1.53-/+0.13, respectively, significantly higher than that in PBS group and CpG ODN group (P<0.05). The serum concentration of IL-2, IL-12 and IFN-gamma in the combined treatment group were 64.6-/+7.4 pg/ml, 314.1-/+26.9 pg/ml and 61.9-/+7.3 pg/ml, respectively, significantly higher than those in the other 3 groups (P<0.05). The tumor formation rate in the combined treatment group was significantly lower than that in MA group (25.0% vs 75.0%, P<0.05). CONCLUSION: CpG ODN can enhance the immunity and decrease the tumor formation rate following a rechallenge with CT26 cells in mice treated with MA.

3,28-Diacet-oxy-29-bromo-betulin.

In the title mol-ecule, C(34)H(53)BrO(4), all the cyclo-hexane rings adopt chair conformations, while the cyclo-pentane ring adopts an envelope conformation. In the crystal, weak inter-molecular C-H⋯O hydrogen bonds link the mol-ecules into corrugated sheets parallel to the ab plane.

[Invertebrate biodiversity in litter layers of natural forest and Eucalyptus plantation in eastern Guangdong, China].

A comparative study was made on the community structure of invertebrates and the species diversity of thrips in the litter layers of natural forest and Eucalyptus urophylla plantation in eastern Guangdong of China. The results showed that in natural forest, Acarina, Collembola, Dipteran larvae, Hymenoptera, Thysanoptera, and Coleoptera were the most abundant invertebrates, accounting for 96.5% of the total individuals collected; while in Eucalyptus plantation, Acarina, Collembola, Dipteran larvae, and Lepidopteran larvae were the dominant invertebrate groups, which accounted for 96.3% of the total. The diversity of invertebrate assemblages was much higher in natural forest than in Eucalyptus plantation, based on the comparsions of Shannon-Wiener diversity index (H'), Pielou eveness index (J), Density-group index (DG), and Simpson dominance index (D). The individuals and species of fungus-feeding thrips were also more abundant in natural forest than in Eucalyptus plantation. However, there was no significant difference in the average density of invertebrates between natural forest and Eucalyptus plantation, because the individuals of Acarina were predominant, constituting 77.6% of the total. All of the results suggested that it is important to remain the understory and litter to improve the litter invertebrate diversity in fast-growing Eucalyptus plantation.

[Transferrin receptor imaging for tracing mesenchymal stem cells implanted in the spinal cord].

OBJECTIVE: To explore the feasibility of tracing mesenchymal stem cells in vivo with scintigraphy. METHODS: Transferrin receptor expression of cultured mesenchymal stem cells (hMSCs) was quantified with radioligand-receptor binding assay before the cells were transplanted into the spinal cord of rabbits. (131)I-labeled transferrin was then administered into the subarachnoid space of the rabbits, and scintigraphic images were acquired with a gamma camera at different time points after the administration. In the control experiments, (131)I-labeled human serum albumin was used in stead of (131)I-transferrin as the tracer, or only PBS was injected without stem cell transplantation. The images were semi-quantitatively analyzed with region of interest (ROI) techniques, and the phosphor imaging on the spinal sections were performed. RESULTS: Radioligand-receptor binding assay showed 10 770 binding sites with high affinity (KD=0.982 nmol/L) for Fe saturated transferrin on each human mesenchymal cell. Visible accumulation of radioactivity at the cell transplantation sites was observed 16 h and 24 h after intrathecal injection of (131)I-transferrin tracer, but not in two control groups. ROI analysis showed that the difference between (131)I-transferrin and the control groups was statistically significant (P<0.05). Phosphor imaging further verified that it was the specific coupling of transferrin to the implanted cells that resulted in radioactivity accumulation at the transplantation sites. CONCLUSIONS: Transferrin receptor imaging is capable of in vivo tracing of the implanted stem cells, and has the potential for use in non-invasive monitoring for stem cell transplantation therapy after further technical improvements.

[D2 receptor expression on immortalized human neural progenitor cell line hNPC-TERT in vitro and in vivo].

OBJECTIVE: To observe D(2) receptor expression on human neural progenitor cell line hNPC-TERT before and after transplantation into rabbit central nervous system. METHODS: D(2) receptor expression on cultured hNPC-TERT cells was verified and quantitatively analyzed with immunofluorescence assay and receptor radio ligand binding assay, respectively. 3 x 10(6) hNPC-TERT cells were implanted in the spinal cord of New Zealand rabbit with HeLa cells as the control. Two days after implantation, positron-emission tomography (PET) scan with (11)C-raclopride as the radiotracer was performed in the living animals or for the isolated spinal cords, and cryosections of the spinal cord containing the implanted cells were prepared for immunofluorescence assay. RESULTS: Cultured hNPC-TERT cells showed high expression of D(2) receptor (Bmax=8 x 10(4)). PET scans of the rabbits identified visible radioactive accumulations at the site where hNPC-TERT cells were implanted but not at the site of HeLa cell implantation. Region of interest analysis showed a significant difference between the two cells in the maximal standard uptake value at the cell implantation sites. The results were further confirmed with ex vivo PET imaging of the spinal cord and tissue immunofluorescence assay. CONCLUSION: Human neural progenitor cells hNPC-TERT highly express dopamine D(2) receptors and retain this capacity after implantation into the spinal cord, suggesting their potential for treatment of such nerve system disease as Parkinson syndrome.

[Tracking neural progenitor cells transplanted into rabbit spinal cord by detection of dopamine receptor 2 with positron emission computed tomography].

OBJECTIVE: To explore the effect of in vivo positron emission computed tomography (PET) in tracking the stem cells transplanted into spinal cord. METHODS: Telomerase-immortalized human neural progenitor cells of the line hNPC-TERT were cultured. HeLa cells were used as control cells. RT-PCR was used to detect the mRNA expression of dopamine receptor 2 (D(2)) in both cell lines. (3)H-raclopride was added into the suspensions of these 2 cell lines. RT-PCR was used to detect the mRNA expression of D(2), and immunofluorescent staining was conducted on the cells to detect the protein expression of D(2). Twenty-two New Zealand rabbits were randomly divided into 4 groups to undergo transplantation of hNPC-TERT cell suspension into the spinal cord at the segment T10, or transplantation of HeLa cells. Two day after the transplantation some rabbits were killed to take out the spinal cord at the segment T10 to undergo immunofluorescent staining to examine the radioactivity in the spinal cord. Some rabbits were injected with (11)C-raclopride intravenously and then underwent PET imaging. RESULTS: RT-PCR showed that mRNA expression was positive in the hNPC-TERT cells but negative in the HeLa cells, and immunofluorescent staining showed protein expression of D(2) in the in the hNPC-TERT cells and not in the HeLa cells. The spinal cords specimens taken 2 days after transplantation had human-specific nuclear (HN) antigen positive and D(2) positive cells. Fluorescent microscopy showed that the hNPC-TERT cells in the injection site did not migrate remarkably; and showed that the D(2) staining was negative and HN antigen was positive in the HeLa cells. (11)C-raclopride PET imaging of the live rabbits showed accumulation of radioactivity at the hNPC-TERT cell injection site with a standard uptake value significantly higher than that of the HeLa cell transplantation group (P < 0.01). (11)C-raclopride PET imaging of the isolated spinal cords showed rounded focal image of increased radioactivity in the hNPC-TERT cell transplantation group and linear image of radioactivity without clear border in the HeLa cell transplantation group. CONCLUSION: PET imaging with as radiotracer targeting at specific cellular marker is effective in tracking cells into the body and in vivo visual evaluation of stem cell transplantation.

[Transferrin receptor expression of human mesenchymal stem cells and in vitro tracking by autoradiography after transplantation in spinal cord].

OBJECTIVE: To determine transferrin receptor (TfR) expression of human mesenchymal stem cells (MSCs) in vitro and after transplantation in rabbit spinal cord,and to detect implanted MSCs by in vitro autoradiography. METHODS: Human mesenchymal stem cells (hMSCs) were isolated from fetal blood. Flow cytometry assay, immuno-fluorescent staining and receptor binding assay were used to determine TfR expression of hMSCs. Radioiodinated transferrin saturated with iron [(125)I-Tf(Fe)(2)] was used as tracer. The hMSCs transplanted in rabbit spinal cord was tracked by in vitro autoradiography. Diffusion of (125)I-Tf(Fe)(2) in spinal cord was examined with autoradiography. RESULTS: TfR expression of MSCs was demonstrated by flow cytometry assay, immuno-fluorescent staining and receptor binding assay in vitro. (125)I-Tf(Fe)(2) bound to hMSCs with a equilibrium dissociation constant (KD) of (0.98+/-0.12) nmol/L and a maximal density of binding sites (B(max)) of (107 702+/-6 226) sites per cell. Immuno-fluorescent staining showed that TfRs were expressed on hMSCs on the 2nd day but not be expressed on the 10th day post transplantation. Autoradiography showed distinct accumulation of (125)I-Tf(Fe)(2) but not (125)I-HSA at hMSCs implantation sites of spinal cord sections on the 2nd day post transplantation. (125)I-Tf(Fe)(2) had diffused into spinal cord 16 hours after incubation. CONCLUSION: Implanted hMSCs could be detected by in vitro autoradiography with (125)I-Tf(Fe)(2) on the 2nd day after being transplanted in spinal cord. To track implanted hMSCs with radionuclide imaging techniques in vivo, TfR was a suitable target for imaging and radioiodinated Tf(Fe)(2) was a feasible tracer.