Effects of oxygen glucose deprivation and reperfusion on gap junction and glucose uptake of astrocytes / 中华行为医学与脑科学杂志

Objective To explore the relationship between gap junction and glucose uptake of astrocytes under oxygen-glucose deprivation(OGD) and reperfusion.Methods Cerebral cortical astrocyte from 1 day newborn SD rats were undergone the primary culture.The ischemia cell model was established by OGD.This experiment were divided into control group,OGD group and OGD + CBX group.After different reperfusion time (0 h,12 h 24 h and 48 h),the glucose uptake of astrocyte was measured by 2-NBDG through flow cytometry analysis and connexin 43(Cx43) gap junction plaques was detected using immunofluorescene.Results Compared with the control group,the glucose uptake of astrocyte was up-regulated induced by OGD following different reperfusion time.The glucose uptake of OGD group was (2.32 ± 0.43)nmol/μgDNA in 24 hours reperfusion time and was (0.95±0.28)nmol/μgDNA in control group.The upregtulation was up to 2.63-fold increase (t=13.99,P=0.0024) in 24 hours after reperfusion.Compared with the control group,the Cx43 gap junction number was up to 2.5-fold increase(t=11.34,P=0.003) and the size was 1.85-fold increase (t=10.27,P=0.004) in 24 h reperfusion.The glucose uptake of astrocyte after OGD was reduced by CBX and the decrease was 42％ in 48 h after reperfusion.Conclusion Those results urges us consider the clinical treatment for interfering with Cx43 gap junction.

Effects of oxygen glucose deprivation and reperfusion on gap junction and glucose uptake of astrocytes / 中华行为医学与脑科学杂志

Objective@#To explore the relationship between gap junction and glucose uptake of astrocytes under oxygen-glucose deprivation(OGD) and reperfusion.@*Methods@#Cerebral cortical astrocyte from 1 day newborn SD rats were undergone the primary culture. The ischemia cell model was established by OGD. This experiment were divided into control group, OGD group and OGD+ CBX group.After different reperfusion time (0 h, 12 h 24 h and 48 h), the glucose uptake of astrocyte was measured by 2-NBDG through flow cytometry analysis and connexin 43(Cx43) gap junction plaques was detected using immunofluorescene.@*Results@#Compared with the control group, the glucose uptake of astrocyte was up-regulated induced by OGD following different reperfusion time.The glucose uptake of OGD group was (2.32±0.43)nmol/μgDNA in 24 hours reperfusion time and was (0.95±0.28)nmol/μgDNA in control group. The up-regulation was up to 2.63-fold increase (t=13.99, P=0.0024) in 24 hours after reperfusion.Compared with the control group, the Cx43 gap junction number was up to 2.5- fold increase(t=11.34, P=0.003) and the size was 1.85-fold increase (t=10.27, P=0.004) in 24 h reperfusion. The glucose uptake of astrocyte after OGD was reduced by CBX and the decrease was 42% in 48 h after reperfusion.@*Conclusion@#Those results urges us consider the clinical treatment for interfering with Cx43 gap junction.

Expression and Clinical Significance of CXCR1 and CXCR2 in Triple-negative Breast Cancer / 中国医科大学学报

Objective To investigate the expression of CXC chemokine receptor 1 (CXCR1) and CXC chemokine receptor 2 (CXCR2) in triplenegative breast cancer (TNBC) and its significance.Methods The EnVision immunohistochemistry system was used to detect the expression of CXCR1 and CXCR2 in 28 specimens from normal tissues adjacent to breast cancer,30 specimens of invasive ductal carcinoma not otherwise specified (IDC-NOS),and 34 TNBC specimens.The relationship between the expression of CXCR1 and CXCR2,and clinical pathological features was analyzed.Results In the adjacent normal tissues of breast cancer,the positive expression rates of CXCR 1 and CXCR2 were 89.3％ and 92.9％,respectively;CXCR1 and CXCR2 expression displayed no significant association with clinicopathological parameters such as age,tumor size,or Scarff-Bloom-Richardson (SBR) score.The positive expression rates of CXCR1 and CXCR2 were 63.3％ and 60.0％,respectively,in the IDC-NOS samples and 23.5％ and 35.3％,respectively,in the TNBC samples.There was a positive correlation between CXCR1 and CXCR2 expression in the TNBC samples (r =0.606,P ＜ 0.001).Conclusion Correlation of CXCR1 and CXCR2 expression with sample type was strong in adjacent normal tissues,moderate in IDC-NOS samples,and low in TNBC samples.CXCR1 expression was positively correlated with CXCR2 expression in all sample types.The expression of CXCR1 or CXCR2 was closely related to the development and promotion of TNBC.

Effects of miR-218-1-3p on Cell Proliferation, Cycle, and Apoptosis of Non-small Cell Lung Cancer / 中国医科大学学报

Objective To investigate the effect of miR-218-1-3p on the proliferation,cycle,and apoptosis of A549 cells in non-small-cell lung cancer.Methods miR-218-1-3p was transfected into non-small cell lung cancer A549 cells by LipofectamineTM 2000 Reagent,and the expression of miR-218-3p was detected by real-time PC R.Invasion and migration were assayed using the Transwell method.The effect of miR-218-1-3p on the proliferation of A549 cells was assayed by the MTS method.Changes in the cell cycle and apoptosis of A549 cells transfected with miR-218-1-3p was detected by flow cytometry.Changes in indicators related to cell proliferation,cycle,and apoptosis were detected by fluorescence quantitative PCR.Results Compared to the control group,the cell proliferation of A549 cells was significantly inhibited (P ＜ 0.05) and the proportion of cells in the S and G2-M phases was significantly decreased when miR-218-1-3p was up-regulated.In addition,compared with the control group,the early apoptotic rate was significantly increased by up-regulating miR-218-1-3p.We further detected indicators related to cell proliferation,cycle,and apoptosis and found that CYCLIN-D1 and BCL-2 were significantly downregulated.Conclusion miR-218-1-3p may inhibit proliferation,induce cell cycle arrest,and promote cell apoptosis of non-small cell lung cancer A549 cells by regulating CYCLIN-D 1 and BCL-2.

Fluid shear stress suppresses TNF-α-induced apoptosis in MC3T3-E1 cells: Involvement of ERK5-AKT-FoxO3a-Bim/FasL signaling pathways.

TNF-α is known to induce osteoblasts apoptosis, whereas mechanical stimulation has been shown to enhance osteoblast survival. In the present study, we found that mechanical stimulation in the form of fluid shear stress (FSS) suppresses TNF-α induced apoptosis in MC3T3-E1 cells. Extracellular signal-regulated kinase 5 (ERK5) is a member of the mitogen-activated protein kinase (MAPK) family that has been implicated in cell survival. We also demonstrated that FSS imposed by flow chamber in vitro leads to a markedly activation of ERK5, which was shown to be protective against TNF-α-induced apoptosis, whereas the transfection of siRNA against ERK5 (ERK5-siRNA) reversed the FSS-medicated anti-apoptotic effects. An initial FSS-mediated activation of ERK5 that phosphorylates AKT to increase its activity, and a following forkhead box O 3a (FoxO3a) was phosphorylated by activated AKT. Phosphorylated FoxO3a is sequestered in the cytoplasm, and prevents it from translocating to nucleus where it can increase the expression of FasL and Bim. The inhibition of AKT-FoxO3a signalings by a PI3K (PI3-kinase)/AKT inhibitor (LY294002) or the transfection of ERK5-siRNA led to the nuclear translocation of non-phosphorylated FoxO3a, and increased the protein expression of FasL and Bim. In addition, the activation of caspase-3 by TNF-α was significantly inhibited by aforementioned FSS-medicated mechanisms. In brief, the activation of ERK5-AKT-FoxO3a signaling pathways by FSS resulted in a decreased expression of FasL and Bim and an inhibition of caspase-3 activation, which exerts a protective effect that prevents osteoblasts from apoptosis.

Fluid shear stress promotes osteoblast proliferation via the Gαq-ERK5 signaling pathway.

Fluid shear stress (FSS) is a ubiquitous mechanical stimulus that potently promotes osteoblast proliferation. Previously, we reported that extracellular signal-regulated kinase 5 (ERK5) is essential for FSS-induced osteoblast proliferation. However, the precise mechanism by which FSS promotes osteoblast proliferation via ERK5 activation is poorly understood. The aim of this study was to determine the critical role of Gαq in FSS-induced ERK5 phosphorylation and osteoblast proliferation, as well as the downstream targets of the Gαq-ERK5 pathway. MC3T3-E1 cells were transfected with 50 nM Gαq siRNA, treated with 5 mM XMD8-92 (a highly selective inhibitor of ERK5 activity), and/or exposed to FSS (12 dyn/cm(2)). Cell proliferation was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The protein expression levels of Gαq, P-ERK5, ERK5, Cyclin B1, and CDK1 were analyzed by Western blot. Physiological FSS exposure for 60 min remarkably promoted MC3T3-E1 cell proliferation, however, this effect was suppressed by siRNA-mediated Gαq knockdown or inhibition of ERK5 activity by XMD8-92 treatment, suggesting that Gαq and ERK5 might modulate FSS-increased osteoblast proliferation. Furthermore, ERK5 phosphorylation was dramatically inhibited by Gαq siRNA. In addition, our study further revealed that FSS treatment of MC3T3-E1 cells for 60 min markedly upregulated the protein expression levels of Cyclin B1 and CDK1, and this increased expression was predominantly blocked by Gαq siRNA or XMD8-92 treatment. We propose that FSS acts on the Gαq-ERK5 signaling pathway to upregulate Cyclin B1 and CDK1 expression, thereby resulting in MC3T3-E1 cell proliferation. Thus, the Gαq-ERK5 signaling pathway may provide useful information regarding the treatment of bone metabolic disease.

Fluid shear stress inhibits TNF-α-induced osteoblast apoptosis via ERK5 signaling pathway.

Fluid shear stress (FSS) is a potent mechanical stimulus and prevents cells from TNF-a-induced apoptosis. Recently, Extracellular-signal-regulated kinase 5 (ERK5) has been found to be involved in regulation of cell survival. However, little is known about the role of ERK5 signaling pathway in FSS-mediated anti-apoptotic effects in osteoblast. In this study, we show that FSS blocks TNF-a-induced apoptosis of MC3T3-E1 cells via ERK5 signaling pathway. We found that physiological FSS for 1 h significantly decreased TNF-α-induced MC3T3-E1 cells apoptosis. After inhibition of ERK5 activity by XMD8-92, a highly-selective inhibitor of ERK5 activity, the ability of FSS to inhibit TNF-α induced apoptosis was significantly decreased. Analysis of anti-apoptotic mechanisms indicated that exposure of MC3T3-E1 cells to FSS for 1 h increased phosphorylation of Bad and inhibited caspase-3 activity. After treatment with XMD8-92, phosphorylation of Bad by FSS was significantly blocked, but caspase-3 activity was increased. In summary, these findings indicated that FSS inhibits TNF-α-mediated signaling events in osteoblast by a mechanism dependent on activation of ERK5, and Bad is a crucial downstream target for ERK5. Those results implied that ERK5 signaling pathway play a crucial role in FSS-mediated anti-apoptotic effect in osteoblast. Thus, ERK5 signaling pathway may be a new drug treatment target of osteoporosis and related bone-wasting diseases.

Effects of corrugated tissue engineered bone scaffold on cell seeding and osteogenesis / 中国组织工程研究

BACKGROUND:The effects of engineered bone scaffold containing seeding cels with different shapes to repair bone defect are varied, while the loaded cellquantity is the important factor influencing the curative effect, but which is rarely reported. OBJECTIVE:By preparing self-made corrugated tissue-engineered bone scaffold and other three forms of bone tissue engineering scaffolds, to study the quantity of loaded cels on different scaffolds and osteogenesis of corrugated tissue-engineered bone scaffold so as to discuss the advantages and features of self-made corrugated tissue-engineered bone scaffold. METHODS: (1) Experimentin vitro: There were four kinds of scaffolds with the same volume and samples: calcium phosphate cement (CPC) corrugated surface scaffold group, smooth surface scaffold group, cylindrical scaffold group and porous cylindrical scaffold with holow tubes group, in which the latter three groups are control ones. A certain volume with same density of rabbit bone marrow mesenchymal stem cels (BMSCs) suspension after osteogenesis induction was seeded onto the scaffolds. After incubation, culture, digestion and colection, cellquantity was counted, absorbance value was finaly detected and cellactivity was proofed by alkaline phosphatase and alizarin red staining. (2) Experimentin vivo: New Zealand rabbits were randomly and equaly divided into recombinant human bone morphogenetic protein-2 (rhBMP-2)/CPC/BMSCs corrugated scaffold group, pure CPC corrugated scaffold group and cancelous bone implant group. Three kinds of scaffold implants with the same volume were inserted into the area between rabbit’s L5, 6 transverse processes bilateraly. At 4, 8, 12 weeks postoperatively, gross and histological observation was performed. RESULTS AND CONCLUSION:(1)Experimentin vitro: The drip of cellsuspension steadily stayed on the surface of corrugated scaffold because of corrugated shape groove and the surface tension of the liquid. The amount of cels per sample digested down from the CPC corrugated surface scaffold was significantly more than that from the other three groups (P 0.05). (2) Experimentin vivo: At each time point the osteogenesis quantity of rhBMP-2/CPC/BMSCs corrugated scaffold group was more than that of the pure CPC corrugated scaffold group (P 0.05). These findings indicate that the characteristics of the self-made corrugated engineered bone scaffold are beneficial to seed cellloading, which supports a large number of osteogenesis and provides feasibility to promote the healing of segmental bone defects.

Application of muscle basal lamina containing neural stem cells in repair of spinal cord injury in rats / 中华创伤杂志

Objective To observe effect of muscle basal lamina containing neural stem cells (NSCs) in repair of spinal cord injury.Methods Thirty-six SD rats from the same nest were used in the study and spinal cord hemisection models were induced.The animals were classified to blank control group (clearance of the lesion edge only with isotonic saline),NSCs group (transplantation of NSCs to the edge),NSCs + muscle basal lamina group (transplantation of complex of NSCs and muscle basal lamina to the edge) according to random number table,with 12 rats per group.At weeks 4 and 8,survival and migration of the transplanted cells and compatibility of muscle basal lamina with the host were detected.At weeks 2,4,and 8,the hindlimb function was assayed using BBB scoring system.Results NSCs in NSCs + muscle basal lamina group grew at the lesion edge,migrated to both sides of the edge,and integrated with peripheral tissues.Whereas,few NSCs survived at the lesion edge in NSCs group and inflammatory cell infiltration was notable.At week 2,there was no statistical difference of BBB score among the three groups.At weeks 4 and 8,BBB score in NSCs + muscle basal lamina group (7.92 ± 1.00,11.38 ± 1.51) was significantly higher than that in blank control group (3.82 ± 0.75,3.71 ± 0.76) and NSCs group (6.25 ±1.06,8.25 ± 1.83) (P＜0.05).Conclusion Muscle basal lamina orients growth of NSCs along its lumen,facilitates migration of host cells to ground substance within its lumen,and reduces local inflammatory reaction.

Effects of exogenous sonic hedgehog on proliferation of neural stem cells in ependymal area after spinal cord injury in adult rats / 中华创伤杂志

Objective To investigate effects of exogenous sonic hedgehog (Shh) on proliferation of neural stem cells (NSCs) in ependymal area and recovery of motor function after spinal cord injury (SCI) in adult rats.Methods Fifty-five female SD rats were involved in the study:five were selected as normal control group and fifty as Shh group (n =25) and SCI group (n =25) after being subjected to SCI at T10 segment using the modified Allen' s method according to the random number table.At 1,3,7,14,and 28 days after operation,restoration of hindlimb motor function of SD rats was assessed with modified Tarlov scale and changes of double positive cells of Brdu and Nestin with double-stained immunofluorescence.Results Tarlov scale revealed statistical difference between Shh and SCI groups since days 7 postoperatively (P ＜ 0.05).In the double-staining test,number of double positive cells of Brdu and Nestin was greater in Shh Group than in SCI Group since day 3 postoperatively [(97.20 ± 18.23) vs (72.60± 15.60),(153.60 ±25.76) vs (112.20 ±23.63),(133.80 ±21.02) vs (94.20± 18.70),(89.80 ± 15.42) vs (43.40 ± 10.62),P ＜ 0.05].Conclusion Exogenous Shh is conducive to the proliferation of ependymal NSCs and the recovery of motor function in SCI rats.

Discussion on origin and prevention of myopia from dysfunction of internal organs and obstruction of channels / 国际中医中药杂志

To explore the origin and prevention of myopia from disfunction of internal organs and obsrtuction of channels.According to the research of ancient literature on relationship between internal organs/channels and myopia,and in the combination with research on neuroelectrophysiology,hemorheology molecular biology and clinic study,the feasibility of treatment on myopia from dysfunction of internal organs and obstruction of channels were analyzed.Dysfunction of internal organs and obstruction of channels played an increasingly important role in the occurring and developing of myopia.In the treatment of myopia,the role of internal organs and channels must be emphasized.

Investigation on diarrhea of 285 HIV/AIDS cases / 国际中医中药杂志

Objective To get overview about diarrhea condition of HIV/AIDS cases among paid blood donors in our country. Methods One-to-one questionnaire was adopted to investigate a total of 285 HIV/AIDS cases among paid blood donors in Henan province. Diarrhea HIV/AIDS patients were further picked out according to 'Criteria of HIV/AIDS diarrhea'and their syndrome characteristics were analyzed. Results There was 41 diarrhea HIV/AIDS patients in all 285 HIV/AIDS cases (14.39%). Deficiency Syndrome and mixed insufficiency and excess syndrome were predominant Syndromes in these patients. Conclusion The rate of diarrhea in HIV/AIDS cases among paid blood donors of our country was lower than some foreign countries. Such internal organs as spleen, liver, kidney, stomach and lung were mainly involved in the disease.

Chemical constituents of Periploca forrestii and their cytotoxicity activity / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of the roots of Periploca forrestii and evaluate their cytotoxicity activities.</p><p><b>METHOD</b>Silica gel column chromatography was employed for the isolation and purification of chemical constituents. The structures were identified on the basis of spectral data and the cytotoxicities of compounds 2-4 were investigated by several tumors cell lines including blood tumor (HL-60, CCRT-CEM), prostate tumor (PC-3, DU-145) and Melanoma (UACC-62).</p><p><b>RESULT</b>Four compounds were isolated and identified as follows, lupeol-20(29)-en-3-nonadecanoate (1), peroiforoside I (2), 3beta,5beta,14beta-3OH-8beta-H-car-20(22)-enolide (3), perplocin (4).</p><p><b>CONCLUSION</b>Compound 1 is a new lupane triterpene fatty acid ester. Compounds 2-4 showed notable cytotoxicity against all tumor lines.</p>

Recent Research of Traditional Chinese Medicine in the Treatment of HIV-related Diarrhoea / 国际中医中药杂志

Objective To get an overview to studies of traditional Chinese medicine treating HIV-related diarrhea recently, and analysis the predominance and inadequacies of these studies. Methods Articles about TCM treating HIV-related diarrhea from 1981 to 2008 in CNKI, CBM, VIP and MEDLINE were summarized and analyzed by analyzing and found, showing the shortages in the methods, detections, and evaluation of treating this disease.

Proliferation of neural progenitor cell after chronic compressive injury of spinal cord / 中国组织工程研究

BACKGROUND: There is still no affirmative conclusion on the proliferative characteristics and the sources of neural progenitor cells after chronic compressive injury of spinal cord in adult mammals and the effects of astrocytes in this process.OBJECTIVE: To investigate the proliferative characteristics and the sources of neural progenitor cell and the effects of astrocytes by means of analyzing the changes of expression of nestin and glial fibrillary acidic protein after chronic compressive injury of spinal cord and after decompression in adult rats.DESIGN: Completely randomized control trial.SETTING: Orthopaedics Research Institute, the Second Hospital of Lanzhou University.MATERIALS: The experiment was completed in Orthopaedics Research Institute of the Second Hospital of Lanzhou University from March to October 2003. A total of 50 adult healthy Wistar rats were selected and randomly divided into normal control group, moderate chronic compressive spinal cord injury group (compressive mass occupied 40% of the diameter of spinal canal), severe compression group (compressive mass occupied 60% of the diameter of spinal canal). Three-day and 10-day decompression groups (depression after 24-hour severe compressive injury) with 10 in each group.MAIN OUTCOME MEASURES: ① Grey value of positive expression of nestin in grey and white matter in spinal cord segment near compression (5 mm to the edge of compression) in rats of each group. ② Expression of glial fibrillary acidic protein in spinal cord of rats in each group.RESULTS: All the 50 rats entered experimental analysis. ①There were significant expressions of nestin in moderate compression group (white matter 235.33±6.48, grey matter 196.28±6.55), severe compression group (white matter 190.45±4.91, grey matter 173.15±5.98), 3-day decompression after severe compressive injury group (white matter 198.39±3.24, grey matter 180.38±4.51) and 10-day decompression group (white matter 202.55±3.54) (P ＜ 0.05), especially in severe compression group (P ＜ 0.01).Compared with the normal control group, the difference between the ex pression of nestin in grey matter and that in ependymal cells on the central canal of spinal cord in 10-day decompression group has no significance (P ＞ 0.05). ②Compared with normal control group, the expression of glial fibrillary acidic protein in spinal cord increased in each injury group,and the amount of positive cells of glial fibrillary acidic protein went up and cell soma was hypertrophic, and the processes became thicker and longer.CONCLUSION: There is neural progenitor cell proliferation in the early stage of chronic compressive injury of spinal cord and after decompression in adult rats. Astrocyte participates in proliferation and migration of neural progenitor cells and has important trophic and repair effects on spinal cord.

Expression of cyclooxypenase-2 and the relation between cyclooxypenase-2 and CD_(105)in osteosarcoma / 肿瘤研究与临床

Objective To study the expression of cyclooxypenase-2(COX-2),prognosis of patients and the correlation between COX-2 and CD_(105)in osteosarcoma.Methods The expression of COX-2 and CD_(105)in human osteosarcoma was detected with immunohistochemistry.Then to count the microvessl density (MVD)were marked with CD_(105),The prognosis of the patients with osteosarcoma was analyzed by Cox multi- variate survival analysis.Results The positive rate of COX-2 expression was 77.5 %,The expression of COX-2 was positively correlated with surgical grade and metastasis of osteosarcoma;There were significant difference between surgical grade Ⅰ and Ⅱ b、grade Ⅰ and Ⅲ(P0.05);Metastatic cases had higher MVD than those without metastasis(P