AIM: To investigate whether Programmed death-1 (PD-1) expression on peripheral CD4~+CD25~(nt/hi)CD127~(lo) regulatory T cells (Treg) was associated with disease progression in HIV-1-infected patients. METHODS: Peripheral blood from 108 HIV-1-infected patients in distinct disease progression statuses and 27 healthy individuals were collected in the present investigation. PBMCs were isolated by centrifugation on Ficoll-Hypaque, followed by staining with anti-CD4-PerCP, anti-CD25-FITC, anti-CD127-PE and anti-PD-1-APC. PD-1 expression on Treg was analyzed by four-color staining flow cytometry. CD4~+T cell absolute counts were determined using Multitest CD3/CD4/CD8/CD45 kit and plasma viral loads were detected on NucliSens EasyQ. All data were analyzed using SPSS14.0 software. RESULTS: In peripheral blood of healthy individuals, Treg expressed PD-1 at very low levels (1.72%±0.65%). In contrast, Treg from HIV-1-infected patients showed a significantly increased PD-1 expression (5.33%±2.24%, P<0.01). Moreover, AIDS patients exhibited statistically higher PD-1 expression on Treg (7.87%±2.23%) than newly HIV-1 infected patients (3.22%±1.01%, P<0.05) and patients in progression to AIDS(5.21%±1.72%, P<0.05). PD-1 up-regulation on Treg was closely correlated with reduced CD4~+T cell absolute counts but elevated plasma viral load. CONCLUSION: Overall, we found that PD-1 expression on peripheral Treg was up-regulated and correlated with disease progression in HIV-1-infected patients for the first time. These findings not only extend our understanding of how Treg functions in HIV-1-infected patients but also support the notion that blocking PD-1/PD-L1 interactions may represent a potential therapeutic strategy for HIV-1-infected patients.

Objective To investigate the effects of different doses of ulinastatin on platelet counts and function after normothermic cardiopulmonary bypass (CPB) in rabbits. Methods Fifty lung-ear white rabbits aged 5-6 months weighing 2.3-3.0 kg were randomly assigned to one of 5 groups (n = 10 each) : control group (group C) and4 ulinastatin groups (group U~1, U_2,U_3,U_4). The rabbits received ulinastatin 1×10~4, 3×10~4, 5×10~4 and 10×10~4 U/kg before CPB in group U~1, U_2, U_3 and U_4 respectively while equal volume of normal saline was given instead of ulinastatin in group C. All rabbits underwent CPB for 30 min at perfusion flow of 72-120 ml·kg~(-1) ·min~(-1). The rectal temperature was maintained at 36.5-37.5℃. Hemodynamic parameters were recorded and blood platelet count, platelet adhesion rate and platelet membrane glycopretein Gp Ⅰ b, Gp Ⅱ b, Gp Ⅲ a receptors were determined before CPB (baseline), at termination of CPB and at 1, 2 and 3 h after CPB. Results The platelet counts were significantly decreased after CPB in all 5 groups (P< 0.05), but there was no significant difference among the 5 groups. The platelet adhesion rates were significantly decreased after CPB as compared with the baseline value before CPB in all 5 groups but the platelet adhesion rates were significantly higher after CPB in group U_4 than in group C. The number of molecules of Gp Ⅰ b, Gp Ⅱ b and Gp Ⅲ a receptors was significantly decreased after CPB in all 5 groups. The number of molecules of Gp Ⅰ b, Gp Ⅱ b and Gp Ⅲ a receptors after CPB was significantly higher in group U_2, U_3 and U_4 than in group C, and there was no significant difference between group U_3 and U_4 . ConclusionUlinastatin 3×10~4-5×10~4 U/kg administered before CPB can inhibit breakdown of platelet membrane glycoprotein receptors. Ulinastatin 10×10~4 U/kg can preserve the platelet adhesion function.

Objective To investigate the role of cAMP-PKA signal transduction pathway in the ischemie pleeonditioning-induced attenuation of ischemia.1eperfusion(ITR)injury in isohted rat hearts.Methods Fifty healthy adult SD rats weighing 300-350 g were anesthetized with intraperitoneal(IP)pentobarbital 50 meg/kg.Their he.arts were excised and perfused in a Langendorff apparatus with 37℃ oxygenated(95%O2-5%CO2)modified K-H solution at a constant pressure of 70 cm H2O,and randomly divided into 5 groups(n=10 each):group Ⅰ I/R;group Ⅱ ischemic preconditioning(IPC);groupⅢH89(PKA inhibitor);group Ⅳ PDTC(NF-κB inhibitor)and group Ⅴ db-cAMP.The experiment started after 10 min stabilization.The isolated hearts were tinct perfuzed for 30 min.followed by 60 min ischemia and 30 min leperfusion in group I/R(Ⅰ).Group IPC(Ⅱ)w88 subjected to 3 episodes of 5 min ischemia at 5 min intervals before I/R.Group Ⅲ and Ⅳ received 5 min perfnsion withH89 10 μmol/L and PDTC 100 μmol/L 3 times at 5 min intervals respectively before I/R.Group Ⅴ was peffnsed with db-cAMP 200 μmol/L for 30 min before I/R.Left ventricular developed pressure(LVDP)and ± dp/dtu were measured at stabilization period and 10, 20, 30 rain of reperfusion. Coronary flow (CF) was measured at stabilization period and 30 min of reperfusion and activities of LDH and creafinc kinase (CK) in the coronary effluent were determined. The myocardial specimens were obtained at 30 rain of reperfusion for determination of NF-κB-DNA binding activity (by EMSA) and expression of TNF-κ mBNA (by RT-PCR ) and p-CBEB (Ser133) (by Western blot). Results Compared with 1/R group, NF-κB-DNA binding activity and TNF-α mRNA expression were significandy decreased, ± dp/dt and CF were significandy increased, CK and LHD activities in the coronary effluent were significantly decreased in group IPC and db-cAMP (group Ⅱ , Ⅴ ) and p-CREB (Ser133) expression was significantly increased in group IPC, PDTC and db-cAMP (group Ⅱ , Ⅳ,Ⅴ ). Compared with IPC group, NF-κB-DNA binding activity and TNF-α mBNA expression were significantly increased, ± dp/dtmax, LVDP and CF were significantly decreased, CK and LDH activities were significantly increased in group H89 and PDTC (group Ⅲ, Ⅳ ) and p-CREB (Ser133) expression was significantly decreased in group H89(group Ⅲ ). Conclusion lschemic preconditioning can attenuate I/R injury in isolated hearts by inhibition of NF-κB-DNA binding activity via cAMP-PKA signal transduction pathway which reduces gene transcription of inflammatory cytokine.

BACKGROUND:Short-segment pedicle screw technology has been extensively used in the treatment and repair of thoracolumbar burst fractures. The technique of operative treatment through the paraspinal muscle approach has advantages such as less trauma and bleeding, and rapid recovery. However, it requires further investigations to verify the superiority of the paraspinal muscle approach of two lateral incisions near the posterior median line. OBJECTIVE:To evaluate the clinical efficacy and Cobb’s angle of short-segment pedicle screw fixation through paraspinal muscle approach of two lateral incisions near the posterior median line in the treatment of thoracic and lumbar fractures. METHODS:From September 2010 to June 2012, 56 patients with thoracic and lumbar fractures were included in the retrospective study, including 42 males and14 females, with an average of 45 years (range 18-59 years). According to the surgical approach, patients were divided into two groups, traditional approach (n=25) and paraspinal muscle approach (n=31). The operative time, intraoperative blood loss, postoperative drainage and postoperative ambulant time in the two groups were observed and compared. The visual analog scale scores at 7 days, 1 month and 6 months postoperatively were recorded. The Cobb’s angles of suffered vertebra were measured preoperatively and at 7 days and 6 months postoperatively. RESULTS AND CONCLUSION:Al patients were fol owed up after internal fixation. The paraspinal muscle approach was superior to traditional approach in the operation time, intraoperative blood loss, postoperative drainage and postoperative ambulant time, and visual analog scale scores at 7 days and 1 month postoperatively (P<0.05). There was no significant difference between the preoperative and postoperative Cobb’s angle in the two groups (P>0.05). The short-segment pedicle screw fixation through paraspinal muscle approach of two lateral incisions near the posterior median line in the treatment of thoracic and lumbar fractures, is an effective and minimal y invasive treatment, with less trauma, less bleeding, rapid recovery, and simple operations. Similar to open surgery, this treatment can recover the anatomical morphology and reconstruct spinal stability, and had good biocompatibility to the host.

Flashing light effect on microalgae could significantly improve the light efficiency and biomass productivity of microalgae. In this paper, the baffles were introduced into the traditional flat plate photobioreactor so as to enhance the flashing light effect of microalgae. Making Chlorella sp. as the model microalgae, the effect of light intensity and inlet velocity on the biomass concentration of Chlorella sp. and light efficiency were evaluated. The results showed that, when the inlet velocity was 0.16 m/s, with the increase of light intensity, the cell dry weight of Chlorella sp. increased and light efficiency decreased. With increasing the inlet velocity, the cell dry weight of Chlorella sp. and light efficiency both increased under the condition of 500 μmol/(m2 x s) light intensity. The cell dry weight of Chlorella sp. cultivated in the novel flat plate photobioreactor was 39.23% higher than that of the traditional one, which showed that the flashing light effect of microalgae could be improved in the flat plate photobioreactor with inclined baffles built-in.
Biomass ; Chlorella ; growth & development ; Culture Techniques ; instrumentation ; Light ; Microalgae ; growth & development ; Photobioreactors

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.

[Objective] To investigate the expression of MICA mRNA and protein in osteosarcoma and give a more comprehensive understanding to its immune evasion.[Methods] RT-PCR was used to analyze MICA mRNA in 11 osteosarcoma tissues and 5 osteosarcoma cell lines.MICA expression was examined in 66 paraffin-embedded osteosarcoma tissues and 6 paraffin-embedded normal bone tissues by immunohistochemistry,and MICA protein in 9 fresh osteosarcoma tissues was detected by Western blot too.MICA surface expression was estimated by flow cytometry.[Results] Nine of eleven (81.8%) osteosarcoma specimens and all of the five cell lines consistently expressed MICA mRNA.Up-regulation of MICA expression was found in osteosarcoma (34/66,51.6%),compared with normal bone tissues (0/6,0%).The cell lines Saos-2,MG63,and HOS showed positive surface MICA expression,while the U2OS and OS732 showed very limited expression.[Conclusion] MICA mRNA and protein predominantly expressed on osteosarcoma tissues and cell lines.

[Objective] To investigate the expression of NF-κB and IL-lra in gastric cancer, and to explore their correlation with the clinicopathological parameters and prognosis of gastric cancer (GC). [Methods] Expression of NF-κB and IL-1ra was detected by immunohistochemistry in tissue microarry of 359 cases of GC. [Results] The expression rates of NF-κB in the patients with metastasis of lymph node, TNM Ⅲ-Ⅳ stage, and poorly differentiated of histology were 80.2%, 80.0%, and 79.2%, respectively. The expression rates of IL-1ra in the patients with tumor size ≤3 cm, early stage, non-metastasis of lymph node, and TNM Ⅰ-Ⅱ stage were 61.7%, 75.0%, 66.4%, and 61.9%, respectively. The 5-year survival rate of the cases with non-expressed NF-κB was statistically higher than that of the cases with expressed NF-κB(P=0.036). The 5-year survival rate of the patients with negative expression of NF-κB and positive expression of IL-1ra was statistically higher than the others (P=0.021). [Conclusions] NF-κB is the adverse predictors of prognosis of gastric cancer. IL-1ra maybe play a protective role in early gastric cancer stage, but it is necessary to study deeply and get more evidences.

Objective To explore the experimental conditions of labeling rabbit bone marrow mesenchymal stem cells (Rb-MSCs) with 5,6-carboxyfluorescein diacetic succinimidyl ester (CFSE) and to investigate the impact on the biological characteristics of Rb-MSCs in vitro.Methods Rb-MSCs were separated and purified by whole bone marrow adherent culture and then were identified by morphology and surface markers.Rb-MSCs were labeled with CFSE and the labeling effect was measured by flow cytometer.The proliferation capacity of labeled cells was detected with CCK-8.The differentiation capacity of labeled cells was investigated by being induced to osteoblasts and lipoblasts.The capacity of labeled cells to secret vascular endothelial growth factor (VEGF) was detected by VEGF ELISA kits.Results The primary Rb-MSCs adhered in 48 h,being fusiform and colony-like growth.Subculture cells became fibroblast-like cells in order with uniform configuration.Most (above 98％) cultured cells expressed the surface markers CD29 and CD44 except for CD45.Compared with other labeling conditions,10μmol/L final concentration of CFSE and 10 min was the best one with a 100％ labeling rate and high fluorescence intensity.Compared with unlabeled cells,the ability of the labeled cells to proliferate and to secrete VEGF was not significantly decreased (P ＞ 0.05).Moreover,the labeled cells had osteogenic and adipogenic differentiation capacity.Conclusions It was a simple and efficient method to label Rb-MSCs with CFSE,especially in a short-term.The capacity of cell proliferation,differentiation,and secretion were not affected.

Objective To evaluate the blood-saving effect of tranexamic acid in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB).Methods The study was a prospective,randomized and placebo-control trial.Two hundred ASA Ⅰ-Ⅳ patients,aged 18-64 yr,weighing 50-100 kg,were randomized to receive placebo (group C,n =100) or tranexamic acid (group T,n =100).Tranexamic acid 10 mg/kg was intravenously infused over 20 min before skin incision followed by continuous infusion at 10 mg· kg-1 · h-1 until the end of operation in group T.While the equal volume of normal saline was given in group C.The total volume of postoperative chest tube drainage,postoperative massive bleeding and a second thoracotomy for stopping the bleeding were reordered.The requirement for transfusion of allogeneic blood and complications during the perioperative period were also recorded.Results Compared with group C,the total volume of postoperative chest tube drainage and incidences of postoperative massive bleeding and a second thoracotomy for stopping the bleeding were significantly decreased,and the requirement for transfusion of allogeneic red blood cells,platelet and fresh frozen plasma was reduced in group D (P ＜ 0.05).There was no significant difference in the incidence of complications between the two groups (P ＜ 0.05).Conclusion Tranexamic acid exerts the blood-saving effect in patients undergoing CABG with CPB and can significantly reduce postoperative bleeding and transfusion of allogeneic blood.