Table of Content

05 January 2014, Volume 34 Issue 1

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NF-κB signal pathway and downstream microRNA activation and expression in xenografted cervical cancer and its adjacent tissues

2014, 34(1):  1-5. 

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Objective To investigate the expression of miR-15b and miR-16 as well as the status of NF-κB signaling in xenografted cervical cancer and its adjacent tissues，which will provide new insight for the diagnosis and treatment of cervical cancer. Methods The xenograft mouse model of cervical cancer was successfully established. Xenografted cancer and its adjacent tissues, normal fat tissues were collected respectively. The expression of miR-15b and miR-16 were detected by real-time PCR. The expression and sublocation of NF-κB(P 65) was analyzed by immunohistochemistry technology. Results HE staining of the xenograft indicated that the model is successful. The miR-15b levels in the cancer are 32.21±3.67, which significant higher than its adjacent tissues25.16±1.86 and normal fat tissue 1.00±0.12 (P<0.05)，adjacent tissues also significant higher than normal fat tissue (P<0.05); miR-16 levels in the cancer are 28.63±2.34,which significant higher than adjacent tissues22.16±1.76 and normal fat tissue 1.00±0.12 (P<0.05)，adjacent tissues also significant higher than normal fat tissue (P<0.05). NF-κB(P 65) was mainly localized in cytosol of normal adipose tissues，whereas in cervical cancer tissues and adjacent non-tumorous tissues，NF-κB(P 65) translocated into the nucleus. Conclusion NF-κB signal pathway and its regulated miR-15b and miR-16 were upregulated significantly in both cancer tissues and its adjacent tissues. The NF-κB signal pathway and its regulated miR-15b and miR-16 were involved in early molecular events of cervical cancer.

Screening the potential early-warning plasma biomarkers specific to cervical precancerous lesion in Uyghur women based on IPA@ bioinformatics analysis

2014, 34(1):  6-10. 

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Objective This study aimed to screen the early-warning plasma biomarkers of cervical intraepithelial neoplasiaⅡ/Ⅲ stage (CINⅡ/Ⅲ) in Uygur women by using Ingenuity Pathway Analysis (IPA), as well as explain the canceration mechanism of cervical cancer. Methods 31 plasma differentially expressed proteins which were analyzed and identified by proteomic techniques were performed by functional annotation, networks analysis, biofuction analysis, canonical pathways analysis and IPA–biomarker? filter analysis using IPA@ online bioinformatics software. Results As the result of IPA@ analysis, when classified according to function, inflammatory response, cell to cell signaling and interaction, cellular growth and proliferation were most frequently identified in CINⅡ/Ⅲ. Acute phase response signaling and JAK/Stat signaling and IL-4 signaling,etc were identified as the canonical pathways that are overrepresented in CINⅡ/Ⅲ. Two plasma proteins (ApoAⅠ and mTOR) as candidate biomarker were screened. Conclusion The proteins ApoAⅠand mTOR could considered as candidated plasma markers of cervical cancer and cervical precancerous lesion.

The relationship of HPV16E6 mutation and Cervical lesions and cervical carcinoma in Han and Uygur in Xinjiang

2014, 34(1):  11-15. 

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Objective To investigate distribution of HPV16E6 mutation and whether there are differences in Han, Uygur cervical cancer(CC) and precancerous lesions.the relationship between the mutation and high incidence of CC in Han, Uygur women. Methods 140case precancerous lesions and cervical carcinoma patients HPV16 genotypes was chosen and HPV16E6 gene was amplified by polymerase chain reaction (PCR). The PCR fragments were sequenced and analyzed. Results There was mutations in 123 cases , mutation rate distribution in Han and Uygur were 47.37%（27/57）and 50%( 33/66) respectively . They were detected mutations D25E、D64E、I73V、H78Y、D113E, L83V, the hot mutation was L83V , rates was 29.82%（17/57）in Han significant lower than 40.90%(27/66) in Uygur, Rates was D25E 19.30%（11/57）in Han significant higher than7.58%(5/66) in Uygur . Rates was D64E 6.1% in Uygur , but have no prevented in Han . Conclusion There was difference between HPV16E6 gene mutation sites and rates in two nations .Our research suggested that the distribution of D64E in Uygur might be associated with high incidence of CC in this nation.

Txnip interference on HG-induced human kidney proximal tubular cell line apoptosis

2014, 34(1):  16-21. 

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Objective To investigate the effect of Txnip interference on high glucose (HG)-induced apoptosis in human kidney proximal tubular cell line (HK-2). Methods Cultured HK-2 were divided into normal glucose group（NG),high glucose group (HG), HG+contol plasmid vector(HG+C) and GH+ VDUP1 shRNA Plasmid (h) (HG+shRNA).Apoptosis of HK-2 was analyzed by DeadEnd? Fluorometric TUNEL System. ROS production was observed by flow cytometry. The expression levels of Txnip, caspase-3, cleaved caspase-3, Bax, Bcl-2, P38 MAPK, P-P38 MAPK and cytochrome c protein were observed by Western blot. The expression levels of Txnip, Bax, Bcl-2mRNA were observed by RT-PCR. Results Compared with normal glucose group (NG), the production of ROS, the number of cell apoptosis, the expression of cleaved caspase-3 and P-P38 MAPK, ratio of Bax/Bcl-2 and the release of cytochrome c from mitochondria to cytoplasm significantly increased in HK-2 in high glucose group (HG)(P<0.05). Txnip interference inhibited HG-induced ROS production, cell apoptosis, expression of cleaved caspase-3 and P-P38 MAPK, ratio of Bax/Bcl-2 and release of cytochrome c from mitochondria to cytoplasm in HK-2(P<0.05). Conclusions: Txnip interference can prevent HG-induced HK-2 apoptosis through decreasing ROS production, preserving mitochondrial function and inhibiting activation of P38 MAPK.

Construction of Recombinant Adenovirus Vector siRARγ and its effect on mouse hepatic progenitor cells differentiation

2014, 34(1):  22-28. 

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Objective To construct and screen out the recombinant adenovirus vector expressing specific siRNA for mouse retinoic acid receptor-γ (RARγ) gene, and to detect the effect of inhibition of RARγ on hepatic differentiation of hepatic progenitor cells from post coitus day 14.5 mouse liver (HP14.5d). Methods Three pairs of siRNA sequence for mouse RARγ gene were designed and annealed in vitro to make double-stranded DNA, then cloned in SfiⅠ digested pSES-HUS vector and recombinated with the backbone vector pAdEasy-1 in E.coli BJ5183 to construct pAd-siRARγ plasmid. Ad-siRARγ was packaged in HEK293 cell line and used to infected HP14.5 cells. Real-time PCR and Western blot were performed to detect the expression of RARγ. 1 μmol/L ATRA was added to induce HP14.5d cells, ALB-driven Gaussian Luciferase (ALB-Gluc) activity assay and PAS staining were carried out to check hepatic differentiation. Results SiRNA fragments were confirmed to be correctly cloned in adenovirus vector by using PCR, endonuclease cutting and gene sequencing. Cloudiness amplification of RFP-positive cells was observed in HEK293 cell line at day 10 of adenovirus package. At 48 h after infection, more than 60% of HP14.5 cells were infected. Among three siRNA, Ad-siRARγ2, 3 effectively inhibited the expression of RARγ. ATRA induction could increase ALB expression and glycogen storage function of HP14.5d cells, However, Ad-siRARγ2,3 infection significantly reversed ATRA induced hepatic differentiation, ALB-Gluc activity and the ratio of PAS stained cells were statistically lower than that in induction group（p<0.05）. Conclusions Successfully constructed and screened out the recombinant adenovirus vector containing siRNA for mouse RARγ gene, which could effectively inhibit the expression of RARβ in HP14.5d cells and then reverse its hepatic differentiation with ATRA treatment.

Effects of AQP1 knock down by RNAi on morphology and water transport in mouse Schwann cells

2014, 34(1):  29-35. 

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Objective To explore the relationship between the expression of AQP1 gene and Schwann cells swelling. Methods The AQP1 expression in Schwann cells of mouse facial nerve tissues was detected by immunofluorescent staining. The RNA interference by lentiviral transduction was used to specifically down-regulate AQP1 expression in Schwann cells. AQP1-shRNA and scr-shRNA transduced cells were observed daily during AQP1-KD by using phase contrast microscopy. Cell volume of scr-shRNA and AQP1 shRNA treated cells was measured daily from the day of treatment, through day 6. Cell viability was measured by MTT assay. Apoptosis effects of AQP1 shRNA on Schwann cells were detected by flow cytometry. Results Schwann cell primary cultures maintained a high level of AQP1 water channels, representing an ideal cell model to study the role of AQP1 in the facial nerve. AQP1 expression in AQP1-shRNA cells was significantly lower compared to cells transduced with only the scr-shRNA by Real-time PCR and Western Blot analysis(P<0.05).AQP1 gene silencing resulted in a cell shrinkage phenotype , as validated by cell volume determinations(P<0.01). MTT assay showed that the cell viability in AQP1-shRNA cells was lower compared to cells transduced with scr-shRNA(P<0.05). Flow cytometry assay showed that AQP1gene silencing could not affect the apoptosis and necrosis rates of Schwann cells. Conclusion AQP1 is an important factor responsible for the fast water transport of cultured Schwann cells; AQP1 inhibition might provide a new therapeutic alternative for the treatment of some forms of facial nerve edema.

The diagnosis,treatment and follow-up of a case of hyperinsulinism/hyperammonaemia syndrome

2014, 34(1):  36-40. 

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Objective To evaluate the efficacy of oral protein tolerance test in the diagnosis and treatment of a case of hyperinsulinism/hyperammonaemia syndrome. Methods Clinical features of a boy with hyperinsulinism/hyperammonaemia syndrome were reported.Result The boy developed hypoglycemia induced by hyperinsulinism after eating proteinaceous test meal in combination with hyperammoniemia. Direct DNA sequence analysis revealed the heterozygous mutation c.978G>A(R269H) in exon 7 of the GDH gene compared to the nomal gene. The oral protein tolerance test was positive and a test was designed to choose a proper ratio between carbohydrate and protein. It revealed that the combination of protein and carbohydrate could be helpful to avoid hypoglycemia. Follow-up showed that dietary and diazoxide therapy could be a useful method to treat it. Conclusions Oral protein tolerance test is a helpful method in the diagnosis of hyperinsulinism/hyperammonaemia syndrome. And as a basic treatment, dietary therapy could effectively control the symptom.

Expressions of OCT4 isoforms in Human embryonic and mesenchymal stem cells

2014, 34(1):  41-46. 

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Objective To compare the expression of OCT4 isoforms in human embryonic stem cells (hESCs) and human mesenchymal stem cells (hMSCs). Methods The characterizations of hESCs and hMSCs were confirmed by RT-PCR, immunofluorescence staining, flow cytometry and in vivo/vitro differentiation assays. The expressions of OCT4 isoforms and regulators that control their transcription (NANOG, SOX2) and translation (LIN28) were quantified by real-time PCR, flow cytometry and Western blot. Results Three OCT4 isoforms mRNA were expressed in both hESC and hMSC. However, their expressions were significantly higher in hESC than in hMSC, especially for OCT4A (p<0.01). At protein level, OCT4A and OCT4B-265aa were translated in hESC while no OCT4 protein could be detected in hMSC. NANOG, SOX2 and LIN28 were highly translated in hESCs while hMSCs were weak positive for SOX2 expression but negative for NANOG and LIN28. Conclusion NANOG, SOX2 and LIN28 regulate the expression of OCT4. The different expression profile of OCT4 isoforms in hESC and hMSC indicates that OCT4 may be one of the important factors resulting in the differences of self-renewal and differentiation potentials of stem cells at different developmental stages.

Ezrin Mediates Neutrophil Elastase-induced MUC5AC Secretion in Human Airway Epithelial Cells

2014, 34(1):  47-52. 

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Objective To explore the role of Ezrin in neutrophil elastase (NE)-induced mucin (MUC)5AC production in human HBE16 airway epithelial cells．Methods The HBE16 airway epithelial cells were cultured，transfected with pEGFP-N1-Ezrin-T567D and pEGFP-N1-Ezrin-T567A， respectively， then each group was treated with 0.5 μmol/L NE．The levels of MUC5AC protein in culture medium，MUC5AC protein in cytoplasm，MUC5AC mRNA, and Ezrin protein in culture cells were detected with ELISA，real time-PCR，and immunofluorescence, respectively．Results There was an obvious increasing of MUC5AC protein production and mRNA expression in cells exposed to NE，with elevation of Ezrin protein，all had significant differences when compared with normal control group．Cell transfected with pEGFP-N1-Ezrin-T567D increased MUC5AC secretion in culture medium，P<0.01，compared with single NE-stimulated cells． Ezrin protein were increased, and manily concentrated in cytoplasmic membranes，P<0.05，compared with single NE-stimulated cells． pEGFP-N1-Ezrin-T567A restrained Ezrin expression in cells and decreased MUC5AC protein in culture medium．Conclusion Ezrin is involved in NE-mediated MUC5AC protein secretion but not MUC5AC mRAN expression in HBE16 cells. The phosphrylation of Thr567 site in Ezrin may play a critical role．

The Expression and Clinical Significance of miR-7 In Peripheral Blood of Patients with Systemic Lupus Erythematosus

2014, 34(1):  53-57. 

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Objective To investigate the expression of microRNA-7(miR-7) in peripheral blood of patients with systemic lupus erythematosus(SLE) and its clinical significance. Methods Twenty patiens with SLE and twenty healthy controls(HCs) matched with age and gender were enrolled in this study. Real time quantitative PCR was used to detect the expression of miR-7 in peripheral blood mononuclear cells(PBMC) ,B cells,T cells and monocyte of SLE patients and HCs. PTEN mRNA expression levels were also evaluated by real time quantitative PCR. Flow cytometer was used to detect the proliferation ability of B cells, which transfected with miR-7 precursor or inhibitor and then elvaluated bioinformatics prediction and dual luciferase report gene assay system were performed to identify miR-7 targets. Results The expression of miR-7 in SLE patients was significantly higher than that in HCs(p<0.01). PTEN was one of target genes of miR-7. The B cell transfected with miR-7 precursor had reduced expression of PTEN and promoted proliferation of B cell（p<0.05）．In contrast, the opposite effect was observed with the miR-7 inhibitor transfection. Conclusions: miR-7 was involved in the pathogenesis of SLE by regulating the PTEN expression and B cell proliferation.It may be a potential therapeutic target for SLE.

Relationship between the promoter methylation of IFITM1 gene and cervical cancer in Xinjiang Uigur women

2014, 34(1):  58-61. 

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To explore the relationship between the gene promoter methylation of ITFTM1 gene and the cervical cancer of HPV18 specific PCR were used to analyze the 40 normal and 40 cervical cancer tissues of Xinjiang Uigur women. The relationship between the methylation and infection was also investigated in 40 cervical cancer tissues. RT-PCR was used to analyze IFITM1 gene mRNA expression in 10 methlation positive and 10 methylation negative cervical tissues. Results The methylation frequency of IFITM1 was 31/40 in cervical cancer, however, it was only 3/40 in normal cervical tissues. The corresponding infection frequency of HPV16 and HPV18 (High-risk HPV, hr-HPV ) is 27/40 and 5/40. IFITM1 gene showed a higher methylation frequency in hr-HPV positive cervix compared with the hr-HPV negative cervix (P<0.05). The IFITM1 gene mRNA expression level was more decreased in the 10 methylation positive cervical tissues than the 10 methylation negative cervical tissues(P<0.01). Conclusion The hypermethylated IFITM1 may be the critical marker of cervical cancer that was related to the hr-HPV and had the potential for the early cervical cancer screening.

JAK2/STAT3 pathway is involved in bilateral chronic constriction injury rat neuropathic pain model

2014, 34(1):  62-67. 

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Objective: To evaluate the contribution of the JAK2/STAT3 pathway to neuropathic pain，we examined the effects of this pathway in a rat model of bilateral chronic constriction injury (bCCI). Methods: A rat model of bCCI was established and 60 rats’ behavior tests were performed on the day before surgery and on day 3、7、14 and 21 after surgery, L4~L6 dorsal spinal cord was harvested at the each time point. RT-PCR and Western blot were performed to explore the activation of JAK2/STAT3 pathway. Results: Pain-related behavioral tests socres in the bCCI rats were significant decreased as compared to the sham-operated and na?ve group at each time point postoperatively (P<0.05). SOCS3 mRNA and STAT3 mRNA significantly increased on day 14, accompanied by JAK2mRNA of with a similar time course. IL-6 mRNA level increased on day 3 and showed statistically significant increases on day 21. Western blot analysis showed that JAK2, P-STAT3, SOCS3 increased at different timepoints. Conclusion：Our results suggest that the JAK2/STAT3 pathway in the spinal dorsal horn was significantly upregulated in a rat bCCI model of neuropathic pain which will open new avenues for therapeutic intervention.

Benzyl propionate nandrolone enhences Nampt expression and insulin secretion in NIT-1 islet cells

2014, 34(1):  68-71. 

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Objective To discuss the effects of nandrolone styrene acrylic acid on islet cells under the condition of oxidative stress for the theoretical basis of the treatment of type 2 diabetes. Methods NIT-1 cell were divided into four groups which in different concentrations of glucose (5.6,11.1,16.7 and 27.6 mmol/L),then treat with 10μg/mL Benzyl propionate nandrolone for 48h. Nampt and FOXO1 protein expression were detected by Western blot assay. Cell cycle was determined by FCM and cell insulin secreted level was measured with radioimmuno-assay. Results Treated with benzyl propionate nandrolone for 48 h,1) G0/G1 phase retardation effect of the cell cycle was relieved(p﹤0.01)when the cell cultured in lower energy, but 2) G2/M block effect of the cell significantly remitted(p﹤0.01)under the condition of high glucose concentration. 3) Benzyl propionate nandrolone can promote Nampt protein expression(p﹤0.05) and promote insulin secretion, but inhibiting the dephosphorylated FOXO1 expression(p﹤0.01). Conclusion Benzyl propionate nandrolone would promote islet NIT-1 cell proliferation and insulin secretion, those effects are closely related to increase Nampt expression inhibit of FOXO1.

NAC relieves the A549 cells injury induced by cigarette smoke extract

2014, 34(1):  72-77. 

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Objective To study the protective role of N-acetylsteine in the A549 cell injury caused by cigarette smoke exact and the expression change of insulin-like growth factor binding protein -3 in this process. Methods MTT assay was used to evaluate cell viability. Cell cycle and apoptotic cell proportion in each group detected respectively by PI and AnnexinV-FITC/PI double-labelled flow cytometry. Intracellular reactive oxygen species (ROS) was estimated by fluorescent indicator H2DCFDA. DAPI was used to observe the nuclear morphology. Furthermore, using real-time quantitative RT-PCR as well as western blot methods,the expression level of IGFBP-3 was detected.Result Compared with control group(0.78±0.03), CSE inhibit cell proliferation significantly(0.55±0.04). NAC can reduce cell injury caused by CSE including restoring the viability of A549 cells(0.67±0.04), attenuating G1 block of cell cycle and significantly reducing the proportion of apoptotic cells and so on. High expression of insulin-like growth factor binding protein-3 (IGFBP-3) in A549 cells treated with CSE was found at both transcriptional and protein levels, and concomitant with the restoration of cell growth after treatment with NAC, down regulation of IGFBP-3 was observed.Conclusion From the present study, it is concluded that NAC can antagonize CSE-induced growth arrest of alveolar epithelial cells and that down regulation of IGFBP-3 probably play an important role in this process.

Expression of ATF4 in mouse endometrium during the estrus cycle

2014, 34(1):  78-81. 

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Objective To investigate the expression pattern of ATF4 in mouse endometrium during the estrus cycle. Methods Reverse transcription polymerase chain reaction (RT- PCR), immunohistochemistry and Western blotting were respectively used to detect the expression of ATF4 mRNA and protein in mouse endometrium during each stage of the estrus cycle. Results The ATF4 mRNA expression in estrus stage was higher than those of the other 3 stages, and there was significant difference between estrus stage and the other 3 groups ( P <0.05) . The prominent expression of ATF4 protein was primarily limited to the glandular epithelium cell and luminal epithelium cell cytoplasm in the endometrium. Expression of ATF4 protein was similar with that of the mRNA. Conclusion ATF4, expressed in endometrium periodically in the estrus cycle, might play an important role in the endometrial cyclical variation during the estrus cycle.

Co-culture of Rabbit chondrocytes and Human umbilical cord mesenchymal stem cells inducse Human umbilical cord mesenchymal stem cells to differentiate into chondrocytes

2014, 34(1):  82-87. 

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Objective To determine if the co-culture of rabbit articular chondrocytes and hUC-MSCs in vitro can affect differentiation of hUC-MSCs into cartilage-like cells, especially chondrocytes, and if so, what the optimal ratio of the two cell types is. Methods To Co-culture rabbit articular chondrocytes and hUC-MSCs at a chondrocyte: hUC-MSCs ratio of 4:1, 3:1, 2:1, 1:1, 1:2, 1:3,1:4 for 21 days and cultured in DMEM high glucose medium. Type Ⅱcollagen (COL2-A1) and glycosaminoglycan (GAG) were analyzed qualitatively by toluidine blue and immunofluorescence technique, respectively. The contents of COL2-A1 and GAG were estimated from the determination of hydroxyproline content and Alcian Blue method separately. The mRNA expressions of GAG and COL2-A1 were assayed by real-time fluorescence quantitative PCR. Results The expression of COL2-A1 and GAG on day 21 was much higher in the 4:1, 2:1, and 1:1 groups than in other the experimental group or the induced hUC-MSCs group. Also on day 21, the expression of COL2-A1 and GAG proteins in the 4:1 group was much higher than that in all other groups. Conclusion The optimal cell ratio in Transwell co-culture system appears to be 1:4 (hUC-MSCs :chondrocytes).

Ursolic acid alleviated the LPS-induced damage in THP-1 cells

2014, 34(1):  88-92. 

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Objective To investigate the alleviating effect and mechanism of ursolic acid against LPS-induced damage in THP-1 cells. Methods After THP-1 cells were exposed to 10 μg/L LPS for 20 h, ursolic acid of different concentrations were added. Cell proliferation was tested by MTT, the expressions of TLR4、MCP-1 and IL-6 mRNA were detected by RT–PCR, enzyme-linked immunosorbent assay(ELISA) was applied to detect the production of monocyte chemoattractant protein1 (MCP-1) and interleukin-6 (IL-6), P65 and Phosphorylation-P65 were detected on protein level using Western blot, at last, the nuclear transcription factor kappa B (NF-kappa B) activity was detected by luciferase report system. Results Compared with control group, LPS group can significantly increase MCP-1, TLR4, IL-6 mRNA and MCP-1, IL-6, P65, Phosphorylation-P65 proteins expression and enhance the NF-κB activity. Compared with LPS group, ursolic acid (1, 5 μmol/L) intervention groups can significantly reduce MCP-1, TLR4, IL-6 mRNA expression and MCP-1, IL-6 proteins expression and weaken the NF-κB activity. Conclusion Ursolic acid could alleviate the LPS-induced damage of THP-1 cells by reducing the NF-κB activity.

Immunization efficacy of a recombinant tuberculosis vaccine of rBCG-IL-12p70-TB10.4

2014, 34(1):  93-97. 

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Objective The recombinant Bacilli Calmette-Guérin (BCG) of rBCG-IL-12p70-TB10.4 successfully constructed was used to vaccinate in BALB/c mice to investigate its immunogenicity, and to prepare groundwork in exploring new vaccines against Mycobacterium tuberculosis (MTB). Methods Fusion gene of IL-12p70-TB10.4 was cloned into multiple cloning sites of plasmid pMV361 to construct recombinant plasmid. The recombinant plasmid was transformed into BCG using Electroportation Generator to construct MTB vaccine of rBCG-12p70-TB10.4 (rBCG-IT) and then was immunized in Balb/c mice. 3, 6, 9, 12weeks after immunization, antibody titers were evaluated by indirect ELSIA, and cellular responses were evaluated by XTT, ratios of CD4+ T and CD8+ T cells were detected by flow cytometry, and IFN-γ production was evaluated by ELISA kit. Results The recombinant vaccine of rBCG-IT was able to induce high levels of specific antibody titers, stronger proliferative responses and greater IFN-γ production, compared with BCG vaccine. Conclusion The recombinant vaccine of rBCG-IT could elicit stronger and more long-lasting humoral and cell-mediated responses compared with BCG vaccine, and was worth exploring its protective efficacy.

Orexin A promote the apoptosis of human gastric cancer cell SGC7901

2014, 34(1):  98-103. 

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Objective To explore the effect of Orexin A on the growth of human gastric cancer cell line SGC7901 and its mechanisms. Methods The SGC7901 cells were treated by Orexin A and SB408124 (an antagonist of Orexin A receptor subtype 1). The MTT assay was used to detect the growth inhibitory rates of Orexin A on human gastric cell SGC7901. The apoptosis was determined by Hoechst 33258 fluorochrome staining and flow cytometric analysis; and we used Western blot to detect the expressions of Bcl-2 Bax Cyt c and caspase-3. Results Orexin A can inhibit the growth of SGC7901 cells. This can be shown by a direct relationship between a longer time for cells to culture when given a higher dose of Orexin A. The strongest inhibitory rate was 65.32%±2.51%, when we used 1.0μmol/L Orexin A treat for 24h（P <0.01）; we can see many apoptotic cells that the nuclear condensation in Orexin A group，While cells that were pretreated with SB408124 showed the number of the native cell was decreased (P<0.05). After treated with Orexin A, the apoptosis rate was 59.52%±1.38%，but when we pretreated with SB408124, the apoptosis rate decrease to 38.96%±1.82%；the expressions of Bax Cyt c and caspase-3 protein were increased, but the expression of Bcl-2 was decreased（P <0.05）. As mentioned above, the inhibitory effects of Orexin A on SGC7901 cell were weakened by the pretreatment with SB408124. Conclusions Orexin A significantly inhibits the growth of SGC7901 cells through inducing cell apoptosis. Its molecular mechanism is associated with OX1R.

Immune-modulatory effect of human adipose derived mesenchymal stem cells on T lymphocytes of patients with aplastic anemia

2014, 34(1):  104-108. 

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Objective To investigate the effect of human adipose derived mesenchymal stems cells (haMSC) on T lymphocytes of aplastic anemia patients (AA) and explore its possible mechanism. Methods HaMSCs were extracted from human adipose tissues．T lymphocytes were isolated from peripheral blood of patients with AA by density gradient centrifμgation．HaMSCs were co-cultured with T-lymphocytes. Inhibitory effect of haMSCs on AA-TLCs was measured by MTT assay, T-bet and GATA-3 levels were examined by realtime PCR and Western blot. The levels of INF-γ, IL-2, IL-4 and IL-10 were detected by ELASA. Results HaMSCs showed inhibitory effect on lymphocyte proliferation of aplastic anemia patients in a time dependent manner: After 3,5 and 7 days’ co-culturing, their inhibitory rates were (34.94±10.11)%，(53.44±6.16)%，(68.13±15.24)%, respectively(p<0.05). The T-bet mRNA and protein levels in MSCs+AA-TLCs group were significantly lower than that in AA-TLCs group, while the GATA-3 mRNA and protein levels were significantly higher than that in AA-TLCs group. The concentrations of Th1 type cytokins INF-γ and IL-2 in MSCs+AA-TLCs group [(8.7±2.7) ng/L and (7.3±1.3) ng/L, respectively] were significantly lower than those in AA-TLCs group [(41.5±3.7)ng/L and (61.5±8.7) ng/L, respectively (p<0.05)]. But the concentrations of Th2 type cytokins IL-4 and IL-10 in MSCs+AA-TLCs group [(88.6±15.2) ng/L and (38.3±11.8) ng/L, respectively] were significantly higher than those in AA-TLCs group [(1.4±0.6) ng/L and (1.3±0.2) ng/L, respectively (p<0.05)]. Conclusion HaMSCs can mediate an immunoregulation effect on T lymphocytes of aplastic anemia patients in vitro，which was related with the depression of Th1-dominant response due to the disorder of T-bet and GATA-3 gene expression.

Maintaining a remifentanil target infusion can improve the quality of recovery profiles during emergence from general anesthesia and tracheal extubation

2014, 34(1):  109-112. 

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Objective To explore whether maintaining low dose remifentanil target controlled infusion (TCI) during emergence from general anesthesia can improve the quality of recovery profiles for the patients under total intravenous anesthesia with propofol and remifentanil. Methods Forty elective scoliosis surgical patients were randomized into two groups: control group (n=20, all anesthetic drugs paused during emergence) and remifentanil group (n=20, maintaining low dose remifentanil target controlled infusion while propofol infusion paused during emergence). Awake time, tracheal extubation time and quality score of recovery status were recorded. Results Awake time and tracheal extubation time were (12.76±3.56) min and (13.98±4.06) min respectively in control group, while it were (13.14±3.87) min and (14.21±4.77) min respectively in remifentanil group. There was no significant difference of awake and tracheal extubation time between two groups. However, the patient number with quality score of recovery status 1/2/3/4/5 in control group were 1/10/5/3/1 respectively, while it were 5/12/3/0/0 respectively in remifentanil group (P<0.05). Conclusions Maintaining low dose remifentanil target controlled infusion during emergence from general anesthesia can improve recovery quality and does not prolong awake time and tracheal extubation time for the patients under total intravenous anesthesia with propofol and remifentanil.

Rapid Determination of Human Papilloma Virus With Loop-mediated Isothermal Amplification

2014, 34(1):  113-114. 

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Pathogenic detection of 2 371 cases of hand-foot-mouth disease in Quanzhou

2014, 34(1):  115-116. 

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Research progresses in therapeutic strategies after TKI resistance

2014, 34(1):  117-120. 

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Acquired resistance to EGFR-TKI after a period of time has become a puzzle in clinic treatment. This review introduced the mechanism of acquired resistance to EGFR-TKI and focused on the recent discovered treatments after TKI resistance, including pan-HER TKI, EGFR-VEGFR double inhibition，C-MET inhibitor, PTEN activator and new drugs targeting EGFR mutation.

Ineffective erythropoiesis and Iron overload

2014, 34(1):  121-124. 

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Iron overload refers to a condition resulting in increased body iron stores, with or without organ dysfunction. Among the secondary iron overload syndromes, besides blood transfusion dependency, a group of anemias that arise from ineffective erythropoiesis may manifest a feature of iron overload. A group of candidate molecules such as hepcidin are identified to serve as vital “singles” in the process. With the up-to-date research achievements of this field, this review mainly states the mechanism and the latest development that how ineffective erythropoiesis cause tissue iron overload.

Progress on elevated gene-1 in astrocyte

2014, 34(1):  125-129. 

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Recently, it has found that the expression of AEG-1 is elevated in subsets of various malignant cells. AEG-1 plays a potential role in several crucial aspects of oncogenesis, including cancer cell surviving, tumor progression, angiogenesis, antiapoptosis, invasion, metastasis and chemoresistance. Meanwhile, it has close relationship with inflammation and natural immunity. AEG-1 functions as a mediator of several signal pathways PI3K/Akt, NF-κB,Wnt/β-catenin. Moreover, it has found that AEG-1 participates in RNA-induced silencing complex mediating gene silencing. Therefore, AEG-1 may serve as an attractive molecular for clinical diagnosis and new anticancer agents target.

NOX4 is a potential therapeutic target against development of pulmonary fibrosis

2014, 34(1):  130-133. 

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Pulmonary fibrosis is a refractory disease with high mortality. The expression of NOX4 mRNA can be activated and increased by TGF-beta1 which could induce the production of a large number of ROS. ROS could further activate Smad which involved in mediating the signaling pathway of TGF-β1/Smad leading to the occurrence of pulmonary fibrosis. Blocking the expression of NOX4 gene through varieties of technologies would disturb the signaling pathway of TGF-β1/Smad which may block the progress of pulmonary fibrosis. Therefore NOX4 may be the most promising therapeutic target in the process of pulmonary fibrosis.

Progress on OLFM4 in common digestive system tumors

2014, 34(1):  134-137. 

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Olfactomedin-domain family 4 (OLFM4) is overexpressed in many human tumors, especially in gastrointestinal malignancies. Many recent studies show that OLFM4 expression in gastrointestinal tumors is significantly higher than that of healthy tissues, while its expression is often downregulated in advanced tumors. Maybe OLFM4 can promote tumorigenesis in the early stage of tumors by its anti-apoptotic effects, the expression loss of OLFM4 is also necessary for tumor progression or metastasis, indicating different biological functions in different pathological stages.

The proceeding research of the expression changes and targets of miR-100 in cancer tissue

2014, 34(1):  138-141. 

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The sequence of miR-100 is highly conserved. In most of the literature about solid tumor, decreased expression of miR-100 were reported, and increase the expression show inhibitory effects on tumor cells. But also, there are reports that over expresstion correlated with tumor recurrence. Still, there are different views on the relationship between the expressions of miR-100 with the pathological staging and disease progression. Several targets have been confirmed, that control the cell proliferation, cell cycle and apoptosis, and regulate the sensitivity of cancer cells to chemotherapy and radiotherapy.

Advantages and Disadvantages of Problem-based Learning (PBL) in Medical Teaching

Xue GAO

2014, 34(1):  142-144. 

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Problem-based learning (PBL) was first proposed by an American professor of neurology Barrows at McMaster University in Canada in 1969. Now it has become a more popular internationally teaching method. It emphasizes cooperation and autonomic learning to solve problems. It is beneficial for stimulating students' learning interest and the problem-solving abilities, however, there are also some drawbacks that will affect embodies of PBL teaching essence.