In recent years,combat missions and non-combat missions of the army have been more and more frequent in the highlands.However,the special geographical environment in highlands will cause a series of damage to the human body,especially cardiovascular systems,respiratory system and hematologic systems,which will threaten the health of soldiers and directly affect the combat effectiveness.Platelets (PLT) are important blood cells.Soldiers entering the highlands always occur changes of PLT levels and functions,in which severe cases can lead to thrombotic disease and thrombocytopenic purpura.Nevertheless,the studies about high altitude-associated PLT abnormalities are scarce and the underlying mechanisms are far from elucidated.Therefore,deep investigations into high altitude-associated PLT abnormalities are the essential steps in preventing PLT-associated diseases,searching for effective intervention measures and improving the life quality of soldiers in highlands.In this article,we will summarize the latest research findings and discuss the potential mechanisms underlying high altitude-associated PLT abnormalities.Finally,we expect to provide new insights into the prevention and treatment of high altitude-associated PLT abnormalities.

Decreased phosphorylation in the insulin signalling pathway is a hallmark of insulin resistance. The causes of this phenomenon are complicated and multifactorial. Recently, genomic analyses have identified ARL15 as a new candidate gene related to diabetes. However, the ARL15 protein function remains unclear. Here, we show that ARL15 is upregulated by insulin stimulation. This effect was impaired in insulin-resistant pathophysiology in TNF-α-treated C2C12 myotubes and in the skeletal muscles of leptin knockout mice. In addition, ARL15 localized to the cytoplasm in the resting state and accumulated in the Golgi apparatus around the nucleus upon insulin stimulation. ARL15 overexpression can enhance the phosphorylation of the key insulin signalling pathway molecules IR, IRS1 and AKT in C2C12 myotubes. Moreover, ARL15 knockdown can also specifically inhibit the phosphorylation of PDPK1 Ser241, thereby reducing PDPK1 activity and its downstream phosphorylation of AKT Thr308. Co-immunoprecipitation assays identified ASAP2 as an ARL15-interacting protein. In conclusion, we have identified that ARL15 acts as an insulin-sensitizing effector molecule to upregulate the phosphorylation of members of the canonical IR/IRS1/PDPK1/AKT insulin pathway by interacting with its GAP ASAP2 and activating PDPK1. This research may provide new insights into GTPase-mediated insulin signalling regulation and facilitate the development of new pharmacotherapeutic targets for insulin sensitization.

Objective To investigate the clinical effects of beraprost sodium hormone therapy with atorv-astatin for diabetic nephropathy in elderly patients.Methods Total of 124 elderly patients with diabetic nephropathy were selected from PLA 324th Hospital during Dec.2012 and Dec.2014,and were randomly divided into observation group and control group,62 patients each.The control group was treated with bera-prost sodium,40 μg per time,3 times a day,oral;the observation group was given beraprost sodium combined with atorvastatin therapy,20 mg per time,1 time a day,oral.Both groups were treated for 8 weeks,and the clinical results of the two groups of patients were compared.Results After treatment,high density lipoprotein cholesterol(HDL-C) of the observation group was higher than the control group[(1.22 ±0.03) mmol/L vs (1.05 ±0.42) mmol/L],low density lipoprotein cholesterol ( LDL-C) was lower than the control group [(1.3 ±0.6) mmol/L vs (2.7 ±1.3) mmol/L],the difference was statistically significant(P <0.05). After treatment,albumin creatinine ratio(ACR) and cystatin C(Cys C) of the observation group were lower than the control group[(6.5 ±2.3) mg/mmol vs (11.6 ±3.2) mg/mmol,(0.8 ±0.3) mg/L vs (1.4 ± 0.6) mg/L],glomerular filtration rate(GFR) was higher than the control group[(138 ±27) mL/min vs (83 ±14) mL/min],the difference was statistically significant(P <0.05).Adverse reaction rate of the observation group was 12.9%(8/62),of the control group was 11.3%(7/62),the difference was not statis-tically significant(P>0.05).Conclusion Beraprost sodium combined with atorvastatin combination thera-py is safe and effective for diabetic nephropathy in elderly patients ,which can significantly improve the clini-cal symptoms and is worth of promotion in clinical practice .

Objectives To investigate the roles of high-mobility group box 1 (HMGB1) protein in lymphangiogenesis and lymphatic node metastasis in colon cancer. Methods Archival tumour specimens from patients with colon cancer were analysed in this retrospective immunohistochemical study. HMGB1, vascular endothelial growth factor C (VEGF-C) and podoplanin protein levels were analysed immunohistochemically . In vitro studies using the colon cancer cell line HCT116 were also undertaken to investigate the relationship between HMGB1, VEGF-C and nuclear factor (NF)-κB. Results Specimens from 70 patients with colon cancer were reviewed. The presence of positive HMGB1 immunohistochemical staining significantly correlated with lymphatic microvessel density, lymph node metastasis and VEGF-C immunohistochemical staining in colon cancer specimens. The presence of positive VEGF-C immunohistochemical staining significantly correlated with lymph node metastasis. The in vitro studies demonstrated that HMGB1 upregulated VEGF-C mRNA and protein in a dose-dependent manner in HCT116 cells, and that this was mediated via NF-κB. Conclusions HMGB1 immunohistochemical staining was significantly associated with lymphangiogenesis and lymphatic node metastasis in colon cancer. There was evidence that HMGB1 upregulates VEGF-C by activating NF-κB in a colon cancer cell line.

目的探索低压暴露对血小板减少性紫癜小鼠模型的外周血细胞免疫功能的影响。方法将BALB/c小鼠随机分为常压生理盐水组（对照组）、常压GP-APS组、低压生理盐水组、低压GP-APS组。于第1、2、4、6d小鼠腹腔注射豚鼠抗小鼠血小板抗体（GP-APS）100ul/20g，造成小鼠慢性持续性血小板减少。于首次注射血清后第3、5、7d后杀鼠取血检测血小板数量及T细胞亚群表达。结果低压生理盐水组和低压GP-APS组较对照组，CD3+和CD4+明显减少（P＜0.001），CD8+轻度下降，CD4+/CD8+比值明显减少。低压GP-APS组或低压生理盐水组较常压GP-APS组，CD3+、CD4+明显低下（P＜0.001），而CD8+改变不明显，因此CD4+/CD8+比值明显下降。低压生理盐水组较低压GP-APS组各项细胞免疫指标均无显著差异（P＞0.05）。而低压GP-APS组的血小板减少分别与低压生理盐水组和常压GP-APS组之间有统计学差异（P＜0.05）。结论高原血小板减少性紫癜小鼠的血小板减少与低氧和细胞免疫功能异常存在一定的关系，而低压暴露对不同免疫状态小鼠的血小板有明显的影响，但细胞免疫损伤可能无叠加效应。

<正>例1男,51岁,因"反复头昏、乏力、尿黄6年,左下肢肿胀2个月,加重伴酱油尿1d"于2009年12月29日入院。2004年无明显诱因出现头昏、乏力、尿黄,血常规提示WBC2.1×109/L,Hb48g/L,PLT76×109/L,MCV122fl、MCH23.5pg,网织红细胞0.20。诊断为"Evans综合征",经激素治疗后好转出院。后因自行停用激素或减量过快而反复出现上述症状,多次住院经

目的:检测糖皮质激素受体在结肠癌组织及细胞株中的表达,探讨地塞米松共处理或预处理24h,结肠癌细胞株对奥沙利铂和氟脲嘧啶化疗敏感性的变化.方法:采用免疫组化检测糖皮质激素受体在61例结肠癌组织标本及4种结肠癌细胞株中的表达;Hoechst33342染色、流式细胞仪检测体外地塞米松对结肠癌细胞株的凋亡诱导作用;以MTT法检测地塞米松共处理或预处理24h后.结肠癌Lovo细胞株对奥沙利铂和氟脲嘧啶化疗敏感性的变化.结果:在57.3%的结肠癌组织标本中糖皮质激素受体呈阳性表达,在四种结肠癌细胞株中仅Lovo和HCT-116表达糖皮质激素受体,HT-29和SW-480细胞不表达.单独应用地塞米松后,对糖皮质激素受体阳性的Lovo和HCT-116细胞有较强的促凋亡作用,对不表达糖皮质激素受体的HT-29和SW-480细胞作用则不明显.以1×10~(-4)mol/L的地塞米松处理Lovo细胞24h,或与奥沙利铂共处理可使奥沙利铂的IC50从(13.7±1.3)μg/mL降低至(5.9±0.6)μg/mL和(4.8±0.7)μg/mL;同样处理可使氟脲嘧啶的IC50从(72.2±8.1)μg/mL降低至(21.1±4.1)μg/mL和(18.6±4.0)μg/mL.结论:部分结肠癌组织及细胞株表达糖皮质激素受体.地基米松体外作用能够促进糖皮质激素受体阳性的结肠癌细胞发生凋亡,与奥沙利铂和氟脲嘧啶预处理或共处理后可以增加结肠癌细胞的化疗敏感性.

患者男,23岁,体重52 kg,血型为O型Rh阳性.确诊慢性髓细胞白血病慢性期3个月.拟行非血缘异基因外周血造血干细胞移植(allO-PBSCT),于2008年11月22日入院,既往史无特殊.供者男性,血型为AB型Rh阳性,HLA-A、B、CW、DRBl、DQBl等位基因位点全相合.预处理方案为经典Bu/Cy方案,采用环孢素(CsA)+短程甲氨蝶呤(MTX)+抗淋巴细胞免疫球蛋白(ATG-Fresenius S)+吗替麦考酚酯胶囊(MMF)预防移植物抗宿主病(GVHD),前列腺素El、熊去氧胆酸、低分子肝素钙预防肝静脉闭锁综合征(HVOD).

Objective To observe the effect of electromagnetic radiation on the activation of JAK2 and STAT5 in the CD34 + cord blood stem cell.Methods CD34 + cord blood stem cells were isolated from the umbilical cord blood.Three exposed groups were administered with 1,5,25 mW/cm~2 electromagnetic exposure for 20 minutes,respectively.In the intervention group with the radiation power of 25 mW/cm~2,AG490 of the final concentration of 10~(-6) mol/L was pretreated for 30 minutes.The cell viabilitv (MTT),the rate of apoptosis (FCM),Caspase-3 protein level (WB) and the activation of JAK2 and STAT5 (WB) were assayed.Results Compared with the normal group,the cell viability of the electromagnetic group decreased at 12 h and 24 h,while apoptosis rate increased with dose.The activation of JA K2 and STAT5 increased at 12 h and 24 h and then decreased.The cell viability of the intervention group was significantly higher than the electromagnetic group with 25 mW/cm~2 irradiation at 12 h and 24 h,and apoptosis rate was significantly decreased as activation of JAK2 and STAT5 were lower. Conclusion JAK2 and STAT5 protein are involved in the process of the electromagnetic radiation within a scope of power (1-25 mW/cm~2) damaging CD34 + cord blood stem cells,suggesting that blocking the activation of JA K2 and STAT5 may be effective to alleviate the cell apoptosis of stem cells.

BACKGROUND/AIMS:Extensive trials have indicated that cancer cells with high glycolytic activity exhibit decreased sensitivity to anticancer agents. Moreover, recent research has proved that a specific inhibitor of hexokinase II, which is a key glycolytic enzyme, may enhance the activity of anticancer drugs. The purpose of this study is to further investigate the effect and mechanisms of hexokinase II on chemosensitivity of colon cancer cells to 5-fluorouracil.METHODOLOGY:The expression of hexokinase II gene was down regulated by RNA interference in colon cancer cell line LoVo, then the IC50 value of 5-fluorouracil to LoVo cells was carried out by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and the protein expression of hexokinase II and thymidylate synthas by Western blot analysis Meanwhile, cell apoptosis and mitochondrial membrane potential were assessed by flow cytometry. Caspase-3 activity was also determined by its substrate color reaction.RESULTS:Down-regulation of the hexokinase II gene of LoVo cells resulted in decreased IC50 value of 5-fluorouracil and increased apoptosis rate, respectively. Furthermore, silencing hexokinase II of LoVo cells induced loss of mitochondrial membrane potential, activation of caspase-3, and inhibition of thymidylate synthase expression.CONCLUSIONS:Our findings suggest targeting hexokinase II has a potential role in the combination 5-fluorouracil treatments of colon cancer.

Objective: To investigate the relationship between Tiam1 and lymphangiogenesis in human colorectal carcinoma (CRC) tissues, as well as the expression of VEGF-C in a CRC cell line (HCT116) after knockdown of the Tiam1 gene with RNA interference (RNAi).Results: In the specimens of CRC tissue, the positivity rate of Tiam1 and VEGF-C was 84% and 58%, respectively. The positivity rate of VEGF-C in the Tiam1 positive group (64.3%) was significantly higher than that in the Tiam1 negative group (25.0%). The LMVD in the Tiam1 positive group (11.35 +/- 3.34) was significantly higher than that in the Tiam1 negative group (7.38 +/- 2.27). In addition, the expression of the Tiam1 gene was efficiently blocked by RNAi. Downregulation of Tiam1 gene expression significantly suppressed HCT116 cell growth in vitro. Compared with untransfected HCT116 cells, HCT116 cells transfected with pGenesil-1-Tiam1 plasmids showed a significant decrease in the expression of VEGF-C.Methods: The expressions of Tiam1, Rac1, VEGF-C and Podoplanin in 50 samples of CRC were detected by immunohistochemical analysis. The lymph microvessel density (LMVD) in Podoplanin positive specimens was evaluated. The results were analyzed statistically to investigate the correlation of Tiam1, VEGF-C, lymph node metastasis and other clinicopathological parameters. An shRNA eukaryotic expression vector against Tiam1 gene was constructed and transfected into HCT116 cells. The expression of Tiam1 gene was assessed by RT-PCR and western blot analysis.Conclusions: We suggest that the Tiam1 gene may act as a crucial therapeutic target for Lymphangiogenesis in CRC.

Objective To investigate the clinical features of the childhood chronic myelogenous leukemia (CCML), including the pathogenesis, incidence, clinical characteristics, diagnostic criterion, prognostic significance and the treatment strategies, etc. Method The data of 148 cases of CCML were comprehensively reviewed and analyzed, and international and domestic literature in the last two decades was reviewed. Results The CCML was found to be rare with unknown etiology, and was an acquired malignant disease of clonal proliferation of hematopoietic stem cells in children. The disease included two clinical types: adult CCML and juvenile CCML. 72.3% of CCML patients were diagnosed as the adult CCML. The clinical feature of CCML consisted of fatigue, low fever, anemia, hepatomegaly, splenomegaly, and lymphadenopathy. The laboratory findings of a typical CCML patient comprised of peripheral blood leukocytosis, basophilia and eosinophilia, myeloid differentiation in different stages, and increased megakaryocytes. The immunohistochemical features of the CCML consisted of highly positive MPO and CD68, significant lowering of neutrophil alkaline phosphatase (NAP), positive for Philadelphia chromosome (Ph) or chimeric BCR/ABL gene, etc. But in most cases of juvenile CCML, the Philadelphia chromosome could not be detected. The Gleevec therapy and hematopoietic stem cell transplantation (HSCT) might give better treatment result for CCML than the traditional therapy. Conclusions CCML has its characteristic clinical feature. The key of good therapeutic result is early diagnosis and treatment. The optimal therapy for CCML is Gleevec regime and HSCT.

Objective To study the effect of HCT116,transfected by Tiam1-SiRNA plasmid,on the lymphangiogenesis of transplantation tumor in nude mouse,and to investigate the relationship between the expression of Tiam1 and lymphangiogenesis.Methods Ten node mice were devided into two groups: Tiam1-SiRNA group and control group.The transfected HCT116 cells and control HCT116 cells were inoculated to nude mice right back subcutaneously.Five weeks later,the transplantation tumors were taken.The expressiong of Tiaml,Racl,VEGF-C,VEGF-D and lymphangiogenesis between the two groups were examined using immunohistochemistry methods(SP) and Western blot.Results Tiam1,Rac1 and VEGF-C/D expression were lower in tranfected tumor than those in control tumor.LMVD in transfected tumor 2.2±1.31 was also reduced than that in control tumor 6.8±2.86.Conclusion Tiam1-SiRNA can inhibited lymphangiogenesis in colorectal cancer of nude mouse by its effect on VEGF-C/D in local microenvironment.

Background: Extensive trials have indicated that cancer cells with high glycolytic activity exhibit decreased sensitivity to anticancer agents. Moreover, recent research has proven that specific inhibitors of hexokinase (HK) II, a key glycolytic enzyme, may enhance the activity of anticancer drugs. The aim of this study was to investigate the effect and mechanisms of HK II on chemosensitivity of a colon cancer cell line (LoVo) to 5-fluorouracil (5-FU). Methods: HK II gene expression was downregulated by RNA interference in the colon cancer cell line LoVo, which was detected by Western blot analysis. Then the IC50 value of 5-FU was determined in LoVo cells via MTT assay. In addition, cell apoptosis and mitochondrial membrane potential (MMP) were assessed by flow cytometry and caspase-3 activity by its substrate color reaction. Results: In LoVo cells, HK II downregulation resulted in a decreased IC50 value of 5-FU and increased apoptosis. Furthermore, HK II downregulation resulted in a decreased MPP and activation of caspase-3. Conclusion: Our findings suggest that targeting HK II may be beneficial for patients with colon cancer treated with 5-FU.

Objective To study the relationship between Tiam1 gene and lymphangiogenesis in colorectal cancer. Methods Tiam1 was silenced by siRNA, then linked with pGenesil-EGFP1, finally transfected into the colorectal cancer cells HCT116. The cells without transfection and transfected with blank vector served as controls. The Tiam1 mRNA expressions were detected by RT-PCR. The expressions of VEGF-C/-D mRNA and protein were detected by RT-PCR and Western blot. Results The expressions of Tiam1 and VEGF-C/-D in HCT116 cells with Tiam1 gene silenced were lower than those in control groups. Conclusion Tiam1 can affect lymphangiogensis in colon cancer by its effect on VEGF-C/-D. It may be a useful marker for lymphatic metastasis in colorectal cancer.

The purpose of this study is to investigate the effect of silencing hexokinase II (HK II) gene with RNA interference (RNAi) technique on colon cancer LoVo cell proliferation in vitro and in vivo. A short hairpin RNA (shRNA) eukaryotic expression vector against HK II gene was constructed, named as plasmid pGenesil-1-HK II, and transfected into LoVo cells. The expression of HK II gene was detected by RT-PCR and Western blot analysis respectively. Then, tumor colony formation was observed, and cell cycle was also assessed by flow cytometry and the contents of intracellular adenosine triphosphate (ATP) by high performance liquid chromatography (HPLC). Furthermore, LoVo cells were injected subcutaneously into nude mice. After a 4-week follow-up period, the sizes and weights of tumors were measured. Moreover, the expression of Ki67 protein was observed by immunohistochemical technique, and cell apoptosis by terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL). Consequently, the expression of HK II gene was efficiently blocked by RNAi. Down-regulation of HK II gene expression significantly suppressed cloning efficiency and cell cycle of LoVo cell in vitro and tumor growth in vivo. Compared with untransfected LoVo cells, LoVo cells transfected with pGenesil-1-HK II plasmids showed significant decrease in the cellular ATP contents and Ki67 expression, and obvious increase in the apoptosis indexes. Our results suggest that HK II gene can act as a crucial therapeutic target for slowing colon cancer growth.

Objective To construct eukaryotic expression vectors of siRNA tageting glucocoriticoid receptorα(GRα),and meas- ure its inhibition efficiency in GRαexpression of the human colon cancer cell line Lovo.Methods Two pairs of siRNA were de- signed and synthesized according to the GRαcDNA sequence in Genebank,and they were then inserted into pGenesil-1 plasmids re- spectively.The recombinants(named pGenesil-1/GRα1 and pGenesil-1/GRα2)were sequenced and identified.The vectors were transfected into Lovo cell line.RT-PCR and Western blot were performed to examine the expression of GRα.Results GRαsiRNA eukaryotic expression vectors were successfully constructed.RT-PCR and Western blot analysis demonstrated that siRNA against GRαreduced the expression of GRαmRNA and protein after pGenesil-1/GRα2 transference.Conclusion Construction of GRαsiR- NA eukaryotic expression vectors lays an experimental foundation for the role of GR during the treatment of colon cancer.

Objective To construct Neuropilins-2 eukaryotic expression vector for RNA interference.Methods Recombinant targeting on gene NRP2 was designed and established with plasmid pGenSil-1 based on NRP2 cDNA equences of Genomes.Two pairs of oligonucleotides were synthesized according to the Tuschl and inserted into plasmid pGenSil-l to generate siRNA eukaryotic expression vector,DH5α strains were transformed,plasmid were extracted,and recombinant vectors were identified by the restriction map and the sequence analysis.The recombinant plasmid(pGenSil-NRP2) was transfected into the cultured LOVO cells.At 48 h after transfection,the whole cell protein was extracted,and the protein level was detected by Western blotting with mouse-anti-human NRP2 monoclonal antibody.Results Recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis.pGenSil-NRP2 expression vector into LOVO cells down-regulated the protein level of NRP2 at 48 h after transfection.The recombinant eukaryotic expression vector were constructed successfully.Conclusion siRNA recombinant can be constructed successfully by RNAi technique for inhibiting NRP2 expression.

OBJECTIVE:To examine the expression of the glucocorticoid receptor and the effect on proliferation and apoptosis induced by Dexamethasone in human colon carcinoma cell lines Lovo, HCT-116,HT-29 and SW-480 in vitro. METHODS: GR protein was determined by immunohistochemistry and GR mRNA by RT-PCR. Proliferation and apoptosis were detected by MTT, DNA Ladder method and flow cytometer. RESULTS: The glucocorticoid receptor was determined in Lovo and HCT-116, not in HT-29 and SW-480. Dexamethasone had an inhibitory effect on the four human colon cancer cell lines, respectively, and Lovo and HCT-116 cells were among the most sensitive. And the characteristic apoptotic DNA Ladders of Lovo were confirmed by DNA Ladder method. Apoptosis cusps were investigated by flow cytometer in Lovo and HCT-116, the rates of apoptosis [Lovo(34.8±1.9)% and HCT-116(33.6±1.4)%] were higher than those [Lovo(2.9±0.4)% and HCT-116(6.4±1.3)%] in the control groups without Dex. CONCLUSION: The results demonstrate expression of the glucocorticoid receptor in Lovo and HCT-116. Inhibiton of proliferation and apoptosis occur on Lovo and HCT-116 cells induced by dexamethasone in vitro, which may have some relationship with glucocorticoid receptor.