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Effect of Recombination Human Erythropoietin on the Apoptosis of Glutamate-induced Retinal Cells

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2014;(2):227-230. doi:10.3870/j.issn.1672-0741.2014.02.024

Objective To study the protective effect of recombination human erythropoietin (rhEPO) on the apoptosis of retinal neurons induced by glutamate.Methods The primary retinal neurons of postnatal SD rats were cultured in vitro for 7 days and divided into 3 groups :control group ,glutamate group and rhEPO pretreatment group.The neurons in the rhEPO pre-treatment group were afterwards allocated to three subgroups in terms of different rhEPO treatments (0.15 ,0.30 or 0.50 U/mL rhEPO for 12 h).Those in glutamate group and rhEPO pretreatment group were treated with glutamate at the concentration of 20μmol/L for 30 min for establishment of the apoptosis model.Twenty-four h later ,the apoptosis index (AI) was assayed by TUNEL and the expressions of BCL-xL mRNA and protein was detected by RT-PCR and immunocytochemistry respective-ly.Results The AI was significantly higher in the glutamate group than in the control group (P<0.01).The AI was signifi-cantly reduced ,and the expression level of BCL-xL mRNA and protein was markedly dose-dependently increased in the rhEPO pretreatment groups compared with the glutamate group (P<0.01).Conclusion The rhEPO pretreatment can inhibit the glu-tamate-induced apoptosis of retinal neurons by up-regulating the expression of BCL-xL .

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Effect of Fasudil on Inflammatory Secretion of Astrocytes

Zhihui DUAN ; Congmin MA

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2014;(2):219-222. doi:10.3870/j.issn.1672-0741.2014.02.022

Objective To observe the effect of Fasudil on inflammatory factors (TNF-α and IL-8) secreted by astro-cytes.Methods Primary rat spinal astrocytes were cultured and randomly divided into three groups :control group ,oxygen-glu-cose deprivation group (OGD group) and Fasudil group.The culture media were harvested at different time points (0 ,3 ,6 ,12 , 24 h) for detection of the concentration of TNF-αand IL-8 by ELISA.Results Compared with the control group ,the concentra-tions of TNF-αand IL-8 in OGD group were significantly increased at each time point (P<0.05);in Fasudil group ,the concen-trations of TNF-αand IL-8 increased at some time points ,but they were much lower than in OGD group (P<0.05) .Conclusion OGD can promote the secretion of TNF-αand IL-8 by astrocytes in vitro ,which can be effectively inhibited by Fasudil.It is suggested that Fasudil plays an important role in regulating the levels of TNF-αand IL-8 secreted by astrocytes.

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Effect of Parecoxib Sodium Preemptive Analgesia on Postoperative Analgesia and Cellular Immune Function in Patients with Uterine Fibroids

Mingyong LI ; Qianqing WANG

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2014;(6):687-691. doi:10.3870/j.issn.1672-0741.2014.06.017

Objective To investigate effects of parecoxib sodium preemptive analgesia on postoperative analgesia and cellu‐lar immune function in patients with uterine fibroids.Methods Totally ,116 cases of patients receiving uterine fibroids surgery were selected from March 2011 to April 2014 in our hospital.All cases were randomly divided into advanced group(n=58)and control group(n=58). The cases in the advanced group were intravenously injected with parecoxib sodium(40 mg)30 min before anesthesia ,while the control group received 5 mL saline intravenously.Postoperative pain conditions of the two groups were e‐valuated by using visual analog scale(VAS method). The PCIA pump first trigger time ,effective compression number and the total amount of fentanyl were recorded. Venous blood was collected before operation ,6 h(T1 ) ,24 h(T2 ) ,48 h(T3 )and 72 h(T4 ) after operation ,respectively. T lymphocyte subsets of CD3+ ,CD4+ and CD8+ and NK cells were detected by using FACS Cali‐bur flow cytometer.The ratio of CD4+ /CD8+ was calculated.Results After 4 ,8 ,12 ,24 and 48 h ,the pain scores of patients were significantly lower in the advanced group than in the control group(all P<0.05). The PCIA pump first trigger time was longer ,effective compression number and the total amount of fentanyl of patients were less in the advanced group than in the control group ,the differences were statistically significant(all P<0.05). At T1 ,T2 and T3 ,CD3+ and CD4+ were significantly higher in the advanced group than in the control group(P<0.05).At T1 and T2 ,CD4+ /CD8+ was significantly higher in the advanced group than in the control group(both P<0.05).At T2 and T3 ,the NK was significantly higher in the advanced group than in the control group(both P<0.05).Conclusion Parecoxib sodium preemptive analgesia used in patients with uterine fi‐broids can reduce postoperative pain ,reduce the amount of analgesics ,reduce the immune suppression ,and improve immune function in patients after surgery.

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Influence of JNK Signaling Pathway in the Epithelial-mesenchymal Transition Process of Human Alveolar Epithelial Cells Induced by TGF-β1

Yong ZOU ; Yulan ZENG

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2014;(6):658-662. doi:10.3870/j.issn.1672-0741.2014.06.010

Objective To explore the role of JNK signaling pathway in epithelial mesenchymal transition (EMT)process of human alveolar epithelial cells A549 induced by TGF‐β1 in vitro.Methods Human alveolar epithelial cells (A549)cultured in vitro were divided randomly into three groups :normal group ,TGF‐β1 group ( treated by TGF‐β1 with 10 ng/mL)and inhibitor group (treated by 10 ng/mL TGF‐β1 and 20 μmol/L Sp600125).Morphological observation on the cells was performed under light microscope after culturing in 3% serum medium for 72 h. The expression of E‐cadherin (E‐cad ,a epithelial cell marker) ,α‐smooth muscle actin (α‐SMA ,a mesenchymal cell marker)and Collagen fibers Ⅰ(ColⅠ ,a mesenchymal cell marker)were tested by RT‐PCR.The level of JNK phosphorylation (p‐JNK)was detected through Western blot.All experiments were repeated three times at least.Results The normal human alveolar epithelial cells (A549)cultured invitro were arranged closely like peb‐bles.E‐cad expressed at a certain level ,while the expression ofα‐SMA ,ColⅠand p‐JNK was weakly detected.In TGF‐β1 group , the cells were spindle‐shaped ,the expression of E‐cad was reduced ,while the expression ofα‐SMA ,ColⅠand p‐JNK was signifi‐cantly increased 72 h after treatment of TGF‐β1. However ,compared with TGF‐β1 group ,spindle‐shaped cells in the inhibitor group were recovered after 72 h being treated by TGF‐β1 and Sp600125 ,the expression of E‐cad was increased and the expres‐sion levels of α‐SMA ,ColⅠand p‐JNK were significantly decreased 72 h after treatment with TGF‐β1 and Sp600125 in inhibitor group. As compared with normal group ,the shape of the cells in inhibitor group was prolate ,and the expression of E‐cad ,α‐SMA ,ColⅠand p‐JNK was not significantly different.Conclusion JNK signaling pathway is related to the process of EMT of human alveolar epithelial cells induced by TGF‐β1. Sp600125 ,a special inhibitor of JNK ,could validly restrain the process.

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Expression and Effect of CGRP in Dorsal Spinal Root Ganglion of Rats with Oxaliplatin-induced Peripheral Neuropathy

Weiting CHENG ; Kai XU

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2014;(6):639-642. doi:10.3870/j.issn.1672-0741.2014.06.006

Objective To observe the pain behavior of rats with Oxaliplatin‐induced peripheral neuropathy(OXIPN)and cal‐citonin gene related peptide(CGRP)expression.Methods Twenty Wistar rats were randomly divided into 2 groups :normal con‐trol group and model group.Oxaliplatin(20 mg/kg)was injected intraperitoneally to the rats in the model group.Mechanical pain threshold and cold allodynia were detected before and 6 ,24 ,72 h and 7 days after modeling ,respectively.Western blot and im‐munohistochemical method were employed to measure the expression of CGRP in dorsal spinal root ganglion of control group and model group.Results The mechanical pain threshold of modal group rats decreased 6 h after injection of Oxaliplatin as compared with the normal group[(11.6 ± 2.2)g vs. (13.9 ± 3.7)g] ,and decreased with prolongation of time ,reduced to the lowest at 72 h[(3.9 ± 1.4)g]and maintained to day 7 ,and the same trend was found in cold chemical allodynia test.CGRP ex‐pression of dorsal root ganglion increased significantly in the model group rats ,and there was a significant positive correlation between the expression of CGRP and the degree of mechanical pain threshold.Conclusion CGRP plays an important role in Ox‐aliplatin‐induced central sensitization of rats with peripheral neuropathy.

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Inhibition of Hepatic Stellate Cell Proliferation and Extracellular Ca2+ Influx by Leptin-siRNA

Xiulan XUE ; Jusheng LIN

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2014;(6):631-635. doi:10.3870/j.issn.1672-0741.2014.06.004

Objective To evaluate the effect of silencing leptin by small interfering RNA(siRNA)on the expression of lep‐tin ,and apoptosis ,proliferation and intracellular Ca2+ concentration([Ca2+ ]i )of hepatic stellate cells(HSCs)and to provide evi‐dence for liver fibrosis gene therapy.Methods HSCs were divided into normal control group ,blank vector group ,siRNA nega‐tive control group and leptin‐siRNA group.After transfection of the leptin‐siRNAs into HSCs ,cell proliferation was measured by MTT assay.Cell cycle and apoptosis were measured by flow cytometry.Expression of leptin was detected by immunocyto‐chemistry and Western blot. [Ca2+ ]i was measured by Fura‐2/AM loading.Results Compared with the normal control group , the blank vector group and the siRNA control group ,the protein expression of leptin and the cell growth were significantly in‐hibited in the leptin‐siRNA group(P<0.05). The proliferation rate of HSCs was significantly different at different time points (24 ,48 and 72 h)(P<0.05).The cell apoptosis rate was increased significantly in the leptin‐siRNA group(P<0.01).At the same time ,Leptin‐siRNA‐induced [Ca2+ ]i was also significantly reduced(P<0.05).Conclusion The leptin gene may play an important role in liver fibrosis progression and is potentially a novel predictive and prognostic marker for liver fibrosis.

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Effects of Goal-directed Volume Therapy on the Intracranial Pressure and the Balance of Cerebral Oxygen Consumption and Supply in Selective Neurosurgery

Shenglan TIAN ; You ZHOU ; Dan FENG

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(1):106-109. doi:10.3870/j.issn.1672-0741.2015.01.021

Objective To investigate the effects of goal‐directed volume therapy (GDVT )on the intracranial pressure(ICP) and the balance of cerebral oxygen consumption and supply in selective neurosurgery. Methods Twenty‐four patients sched‐uled for intracranial tumor resection were randomly divided into 2 groups:conventional fluid management group (group C ,n=12) and GDVT group(group G ,n=12). Patients in group C received introperative fluid transfusion according to classical fluid management strategies while those in group G received GDT according to stroke volume variation (SVV) ,guided by Flotrac‐Vigileo system.Mean arterial pressure(MAP) ,heart rate(HR) ,cardiac index(CI) ,ICP ,SVV and jugular bulb oxygen saturation (SjvO2 )were recorded before the anesthesia induction(T1 ) ,at the moment of intubation(T2 ) ,at the moment of opening the hard meninges(T3),1hafteropeningthehardmeninges(T4),andattheendofthesurgery(T5).Thecerebraloxygenextractionra‐tio(CERO2 )was calculated. The duration of surgery ,crystalloid volume ,colloid volume ,blood transfusion volume ,urinary output and bleeding volume were recorded as well.Results The colloid transfusion volume ,the total fluid transfusion volume and uri‐nary output were significantly increased in group G when compared with those in group C (P<0.05).MAP ,CI ,and SjvO2 were much higher and CERO2 were much lower at T4 and T5 in group G than in group C(P<0.05). There was no significant differ‐ence in the ICP at each time point between groups G and C (P>0.05).Conclusion Goal‐directed fluid therapy optimizes the cardiac preload without increasing the ICP in selective neurosurgery ,and it also improves the balance of cerebral oxygen con‐sumption and supply.

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Extracorporeal Liver Support with Less Fresh Frozen Plasma for Treatment of Acute-on-chronic Liver Failure

Lisha ZHANG ; Manzhi ZHAO ; Dong XU

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(1):87-91. doi:10.3870/j.issn.1672-0741.2015.01.017

Objective To observe the efficacy of extracorporeal liver support by using less fresh frozen plasma in the treat‐ment of acute‐on‐chronic liver failure.Methods A total of 45 patients with acute‐on‐chronic liver failure were divided into ob‐servation group[plasma perfusion(PP) with a small amount of plasma+ plasma exchange(PE)] ,control group 1(PE) ,control group 2(PP+PE)in terms of the amount of plasma used on the day of treatment. All the patients received artificial liver treatnts 62 times totally.Results The clinical symptoms were improved in the three groups after treatments.There were significant differences in the decrease of alanine transaminase (ALT) ,aspartate transaminase(AST) and direct bilirubin(DBil)rather than the decrease of total bilirubin(TBil)and blood ammonia among the groups.No significant difference was noted in the liver and kidney function among the three groups. The improvement of the coagulation function was poor in the observation group when compared with the control group 1 and control group 2 and there were significant differences.Conclusion During the short sup‐ply of the plasma ,plasma perfusion combined with small amount of plasma can be considered to be used in artificial liver treat‐ments ,which can effectively decrease the level of TBil ,relieve symptoms and decrease the occurrence of complications.

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Expressions of SOCS3 and IL-6 in Triple-negative Breast Cancer and Their Chinical Significance

Lin YANG ; Siqi HAN

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(1):82-86. doi:10.3870/j.issn.1672-0741.2015.01.016

Objective To investigate the expressions of SOCS3 and IL‐6 in triple‐negative breast cancer(TNBC)and their clinical significance.Methods The expressions of SOCS3 and IL‐6 in TNBC and normal breast cells were detected by Western blot and fluorescence quantitative PCR.Immunohistochemical staining was used to detect their expressions in 84 TNBCs ,33 breast fibromas and 11 normal breast tissues. The correlation of their expressions in TNBC and the relationship between their expressions and clinical pathological parameters were analyzed as well.Results SOCS3 was not expressed or down‐expressed in TNBC cell lines while IL‐6 was over‐expressed. The positive rate of SOCS3 protein was 32.1% in TNBCs ,63.6% in breast fi‐bromas(P<0.01)and 72.7% in normal breast tissues (P< 0.01);and that of IL‐6 protein was 72.6% in TNBCs ,30.3% in breast fibromas(P< 0.01)and 27.3% in normal breast tissues (P< 0.01).Moreover ,a significant negative correlation was found between the expression of SOCS3 and that of IL‐6 in TNBC(rs = -0.590 ,P<0.01). The expressions of SOCS3 and IL‐6 were markedly associated with tumor size ,lymph node metastasis ,clinical stages and expressions of Ki‐67 and p53(P<0.05) , but not with age ,histological grades or menopausal status(P>0.05).Conclusion The expressions of SOCS3 and IL‐6 are asso‐ciated with the development of TNBC ,suggesting that SOCS3 and IL‐6 can serve as biological markers for evaluating the prog‐nosis ,and therapeutic targets of T NBC.

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Stem Cell Tracking with Amphiphilic Polyethyleneimine Modified Quantum Dots as Functional Optic Probes

Qing WEN ; Renkai LIU ; Zhiyong WANG

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(1):28-31. doi:10.3870/j.issn.1672-0741.2015.01.005

Objective Stem cell therapy has been extensively used in clinical practice. However ,the methods available are unable to non‐invasively detect the distribution of cells in vivo. This study was aimed to establish a novel molecular imaging probe for stem cell labeling ,which would monitor cells via fluorescence imaging. Methods 1‐iodododecane was used to modify polyethyleneimine (PEI) to form amphiphilic macromolecules. These macromolecules encapsulated the hydrophobic quantum dots by self‐assembly to form imaging probes. Results Strong fluorescence signals of the probe occurred at 630 nm ,and the probe could be effectively endocyted by mesenchyme stem cells. In vivo imaging showed that significant optic signals occurred in probe‐labeled mesenchyme stem cells compared with the control cells (blank group:1.47e8 [p/s/cm2/sr]/[μW/cm2 ];test group:2.78e8 [p/s/cm2/sr]/[μW/cm2 ]). Conclusion The nanoprobes have good optic imaging effects ,and can be used to la‐bel cells.

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