Bone Metabolic Markers in Patients with Obstructive Sleep Apnea Syndrome / 中华医学杂志(英文版)

Background@#Obstructive sleep apnea syndrome (OSAS) is prevalent in obesity and is associated with many metabolic abnormalities. The relationship between OSAS and bone metabolism is still unclear. The aim of this study was to investigate the relationship between the severity of OSAS and bone metabolic markers.@*Methods@#A total of 119 obese males were enrolled in this study in spring months from 2015 to 2017. All candidates underwent polysomnography, and their bone mineral density (BMD) and the serum levels of total procollagen type 1 N-terminal propeptide (t-P1NP), N-terminal midfragment of osteocalcin (N-MID), β-C-terminal telopeptide of type 1 collagen (β-CTX), vitamin D (VD), and parathyroid hormone (PTH) were measured. The analysis of variance and Pearson correlation analysis were performed for data analyses.@*Results@#No significant differences in the mean values of BMD were observed among the obesity, mild-to-moderate OSAS, and severe OSAS groups; and the serum levels of t-P1NP and β-CTX in the severe OSAS group were significantly higher than those in the obesity group (48.42 ± 23.78 ng/ml vs. 31.98 ± 9.85 ng/ml, P < 0.001; 0.53 ± 0.24 ng/ml vs. 0.41 ± 0.13 ng/ml, P = 0.011, respectively). The serum level of VD in the obesity group was significantly higher than those in the mild-to-moderate and severe OSAS groups (both P < 0.001), and decreased as the severity of OSAS increased (P < 0.001). The serum level of PTH in the severe OSAS group was significantly higher than those in the obesity and mild-to-moderate OSAS groups (both P < 0.001). The results of correlation analysis indicated that the level of apnea-hypopnea index (AHI) was correlated with the levels of t-P1NP (r = 0.396, P < 0.001), VD (r = -0.404, P < 0.001), and PTH (r = 0.400, P < 0.001), whereas the level of minimum Osaturation (SaOmin) was correlated with the levels of VD (r = 0.258, P = 0.016) and PTH (r = -0.376, P < 0.001).@*Conclusions@#The levels of bone resorption and formation markers in patients with severe OSAS were significantly increased compared to obese men, and the severity of OSAS was correlated with the serum levels of t-P1NP, VD, and PTH.

Preliminary study of the inhibitory effect and mechanism of B16F10-ESAT-6-gpi/IL-21 vaccine on the pulmonary metastasis in mouse models of melanoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanism of B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine on pulmonary metastasis in mouse model of melanoma.</p><p><b>METHODS</b>Twelve 8-week old female C57BL/6 mice were used in this study. The mice were injected with wild-type B16F10 cells through tail vein after immunization with B16F10-ESAT-6-gpi/IL-21 tumor cell vaccine, and the pulmonary metastasis was observed. The CD4(+) and CD8(+) T cells were isolated by magnetic activated cell sorting, and then used for the detection of CFSE/7-AAD cytotoxicity by flow cytometry. Serum from the mice immunized with tumor-cell vaccine was used to detect IFN-γ expression by ELISA. The expression of TGF-β2, ZEB1, E-cadherin, and N-cadherin of tumor tissues was detected by RT-PCR and immunofluorescence, respectively.</p><p><b>RESULTS</b>The mice vaccinated with B16F10-ESAT-6-gpi/IL-21 had significantly fewer nodules in the lung and lower lung weight [(285.8 ± 19.01) mg vs. (406.3 ± 27.12) mg], with lower levels of TGF-β2, ZEB1 and N-cadherin proteins but higher level of E-cadherin protein within the tumor tissue, as compared with the control mice. Meanwhile, the immunized mice had significantly increased CD8(+) T cell killing activity [(42.62 ± 3.465)% vs. (22.29 ± 1.804)%] and IFN-γ expression level [(55.200 ± 7.173) pg/ml vs. (6.435 ± 1.339) pg/ml] over the control mice.</p><p><b>CONCLUSIONS</b>The B16F10-ESAT-6-gpi/IL-21 vaccine can inhibit the metastasis of melanoma in the lung in vaccinated melanoma-bearing mice. This inhibitory effect is associated with CD8(+) T cell immune response and a higher level of IFN-γ, which may influence on the mesenchymal-epithelial transition of tumor cells.</p>

Study on anti ovarian cancer efficacy elicited by CD34~+ umbilical cord blood hematopoietic stem cells transfected with IL-21 in nude mouse model / 中华微生物学和免疫学杂志

Objective To investigate the CD34~+ umbilical cord blood hematopoietic stem cells (CD34~+ UBSC) transfected with interleukin 21 (IL-21) against the ovarian cancer effect in tumor-bearing nude mice. Methods CD34~+ UBSC were obtained from the UBSC by a magnetically activated cell sorting technique and then transfected with recombinant plasmid plRES2-IL-21-EGFP after the CD34~+ UBSC were proliferated in vitro. The therapeutic effect was evaluated by the size of the tumor and the life span in nude mice treated with the CD34~+ UBSC-IL-21. The expression of IL-21 and its bioactivity in CD34~+ UBSC-IL-21 and in local neoplasitc tissues were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR), immune fluorescence technique, ELISA, Western blot, immunohistochemistry and splenocyte proliferative activity. The NK cell cytotoxicity and the numbers of NK cells, serum level of IFN-γ and TNF-αwere simultaneouly detected by FCM and ELISA, respectively. Results CD34~+ UBSC were cultured and transfected with pIRES2-IL-21-EGFP successfully. CD34~+ UBSC-IL-21 could inhibit the tumor growth and extended nude mice life span compared with other groups (P < 0.01). The expression of IL-21 in the neo-plastic tissue, serum level of IFN-γ and TNF-α , NK cell activity and the numbers of NK cells of mice origin and of human origin in splenocytes were increased significantly in the nude mice treated with CD34~+ UBSC-IL-21 compared with other groups(P <0.01). Conclusion The CD34~+ UBSC transfected with IL-21 have competent against ovarian cancer in tumor-bearing nude mice. The findings may establish a foundation for gene therapy of the ovarian cancer by CD34~+ UBSC-IL-21 in clinic application.

Investigation on the anti-tumor effect and mechanisms of the glycosyl phosphatidylinositol modified routine IL-21 tumor vaccine / 中华微生物学和免疫学杂志

Objective To construct the murine IL-21 (mIL-21) tumor vaccine modified by glyco-syl phosphafidylinositol(GPI), and to evaluate its anti-tumor effect and mechanisms. Methods The IL-21-GPI gene was acquired by overlap PCR and inserted into PeDNA3.1. The recombinant plnsmid pcDNA3.1/ IL-21-GPI was transformed into cell B16F10, and the expression of mIL-21 on cell membrane was deter-mined by cell indirect immumofluorescence and flow cytometry (FCM). The bioactivity of mIL-21 was iden-tiffed according to its effects on the proliferation of mouse spleen cells. The anti-tumor effect was evaluated depending on the tumor size and the survival of tumor-beating mice after the tumor vaccine was inoculated into C57BL/6 mice. And the activity of cell-mediated immunity in immunized mice was detected at the same time. Results The recombinant plasmid pcDNA3.1/IL-21-GPI was correctly constructed, which could ex-press mIL-21 binding the membrane with good bioactivity. The vaccine had good anti-tumor effect, and the cell-mediated immunity had been improved in immunized mice. Conclusion The GPI modified mIL-21 tumor vaccine with anti-tumor activity was constructed successfully, which provided a good foundation for studying anti-tumor immunity and therapy in future.

Anti-tumor mechanisms of lymphopenic mice transferred with tumor-specific lymphocytes and immunized with mIL-21 tumor vaccine / 中华微生物学和免疫学杂志

Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.

Gene expression profiles on three kinds of genotype hepatitis C virus core protein in Huh-7 cell line with microarray analysis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze three kinds of genotype hepatitis C virus (HCV) core protein expressed in human hepatoma (Huh-7) cell line and to recognize HCV core proteins biological function and its pathogenic mechanism.</p><p><b>METHODS</b>The Huh-7 cell expressed three kinds of core proteins were established respectively. Affymetrix human gene chip was used for identifying the gene expression dependently on Affymetrix's protocol. All genes changed by 3 or 1.5 folds between the transfected cells and a control cells were further analyzed, and annotated by using NetAffx analysis through Affymetrix website and were categorized based on their biological processes.</p><p><b>RESULTS</b>The HCV-1b core protein caused 16 genes up/down-regulated expression, of which the immune response genes of PF4V1 and SPP1 were up-regulated 3.4 or 4.4 folds respectively. The HCV-2a core protein had caused the immune response gene CXCL5 and apoptosis gene BTF a down-regulated expression of 3.4 and 3.1 folds respectively, but caused the apoptosis genes of HRK and LZTS1 an up-regulated expression of 3.2 and 3.4 folds respectively. As compared with HCV 1b or 2a core protein, HCV-4b core protein caused 111 genes expression changing and it had more obvious effects on gene expression. If we applied 1.5 fold change for a comparison gene expression, a few of the same gene expression profiles might be caused by these two core proteins.</p><p><b>CONCLUSION</b>The three kinds of HCV core protein should have its own expression character and be mainly shown in immune responses, signal transduction, apoptosis, etc. It should be helpful for our recognizing the HCV core protein biological function and its pathogenic mechanism.</p>

Experimental study of poly-DL-lactic acid membrane guided bone regeneration in rabbit radii bone defects / 生物医学工程学杂志

This study was conducted to observe bone regeneration guided by poly-DL-latic acid (PDLLA) membrane in rabbit radii bone defects and to explore the mechanism of the membrane guided bone regeneration (MGBR). The animal models of bony and periosteous defects were established in both radii of 40 adult New Zealand white rabbits. The left defect as the experimental side was bridged with PDLLA membrane tube, the right side as the controlled side was untreated. The specimens were collected at 2, 4, 8 and 12 weeks postoperatively. General observation, X-ray, histological observation and biomechanical examination were applied to the repair of the models of MGBR in both groups. Two weeks after operation, with much new bony callus formed outside the tube at both fragments, the membrane tube covered with connective tissues was filled with haematoma and fibrous callus. Twelve weeks after operation, the PDLLA membrane became white and its tube shape was still maintained. However, new bone callus outside the tube almost completely disappeared, and inside the tubes all radii bone defects were successfully repaired with bony union. On the controlled sides, bone defects were filled with connective tissues 2 weeks postoperatively. And 12 weeks after operation, the typical nonunion that had been formed after bone marrow canals were sealed with cortical bone. On the experimental side, the strength of the newly formed bone at the 12th week was higher than that at the 8th week (P<0.05), whereas the biomechanical examination could not be done on the controlled side. Therefore, these findings suggested that the bone regeneration could be successfully guided by PDLLA membrane, and this MGBR technique might be generally used in the treatment of bone defects and nonunion.

Advance in mechanism study of multi-drug resistance mediated by ABCC1 and its reverse / 医学研究生学报

Multi-drug resistance(MDR) is the most frequent reason for tumor chemotherapeutic failure.ATP binding cassette(ABC) drug transporters play an important role in tumor MDR.ABCC1 is one of the members of ABC transporter super-family,which lead to drug risistance via discarding drug out of cell or changing the drug distribution in a cell.In this article,we reviewed the advance in MDR mediated by ABCC1 and its reverse.

DIFFERENT FACTORS AFFECTING ANTIBODY RESPONSES IN MICE IMMUNIZED BY GENE RECOMBINANT OF HCV STRUCTURE REGION / 免疫学杂志

To seek the optimum experiment methods of animal immunization with HCV gene and to explore the effect on antibody responses in mice immunized by pCD-HCV1 recombinant in different administration, recombinant pCD-HCV1 was constructed by technique of molecular biology and was injected into muscles of Balb/c of mice with different times, routes and dosage of inoculations as well as different treatment. The results showed that the serum antibody level reached 0.183±0.06,0.428±0.05,0.707±0.08 and 0.773±0.07(OD410 value) respectively after recombinant pCD-HCV1(100μg/mouse) were injected into mice once, twice, three times and four times. The antibody level of mice (n＝12) with four times inoculation was the highest; pCD-HCV1 was perfused into stomach orally in mice or were into mice by i.p, s.c and i.m(100μg/mouse, three times) in different routes (n＝6), and the antibody levels were 0.138±0.05, 0.178±0.07, 0.233±0.08 and 0.691±0.05 respectively; after the mice (n＝8) were inoculated with the pCD-HCV1 of different dosage(10μg, 50μg and 100μg) the antibody levels of three groups were 0.11±0.09, 0.33±0.04, and 0.700±0.07, and the results showed a significant difference (P＜0.01); Mice was injected with procaine (100μl, 0.4mg) by i.m or s.c. Then pCD-HCV1 was injected into mice and antibody levels were higher than that of mice immunized directly with recombinant pCD-HCV1 of same dosage. The results may provide a reference data deserved for screening the optimum immunization method of development HCV-DNA-based vaccine in mice model.

Investigation on Key Aspects of the Bilingual Teaching in Microbiology / 微生物学通报

Fundamental specialized courses are important due to their connection between fundamental knowledge and major knowledge,which are regarded as the difficult courses for undergraduates to learn.The useful approaches and experience of bilingual teaching in microbiology,one of the fundamental specialized courses for the students majoring in Bioengineering,are presented here.The key aspects such as choosing textbooks,teaching manners and teaching contents are also discussed.This paper also proposes how to improve the quality of bilingual teaching during the fundamental specialized courses.