The early phase of human sepsis is characterized by a combination of apoptosis and proliferation of T cells

Purpose

T cell activation as well as unresponsiveness has been described in separate studies in sepsis. Our aim was to establish the coexistence of both T cell fate in human sepsis.

Patients and Methods

This is a cross-sectional study of 48 patients presenting with severe sepsis or septic shock and 15 healthy controls. Cytofluorometric techniques were used to quantify T cell activation, apoptosis, proliferation, expression of costimulatory molecules, and cytokine secretion.

Results

Patients with sepsis were characterized by a significant increase in the percentage of activated T cell subsets, as measured using CD69 marker, compared with healthy controls (P < .05). T cell proliferation as measured through Ki67 expression was obvious in infected patients for both CD4 and CD8 T cell subsets compared with controls (P ≤ .006). T cell subset apoptosis as measured using Hoechst dye was also increased in infected patients compared with controls (P ≤ .002). CD4 T cell proliferation was correlated with interleukin 2 secretion (R² = 0.84, P < .001), whereas up-regulation of CD4 T cell apoptosis was correlated with CTLA-4 expression (R² = 0.24, P = .001). No such similar relationship was observed for CD8⁺ T cells.

Conclusions

Concomitant T cell proliferation and T cell apoptosis are observed in human sepsis, being related to a different pathway.     

CDCA3在食管癌中的表达及其对食管癌细胞增殖与凋亡的影响

目的研究细胞周期相关因子(CDCA3)在食管癌组织和细胞中的表达及其对食管癌细胞增殖与凋亡的影响。方法实时荧光定量PCR(qPCR)检测50例食管癌组织及其配对的癌旁组织中CDCA3 mRNA的表达水平,同时检测CDCA3 mRNA在不同食管癌细胞系(Eca109、Kyse-170、TE1)和人食管鳞状上皮细胞系Het-1A中的表达水平;采用si-CDCA3-1、si-CDCA3-2转染Eca109和TE1细胞,另转染si-NC作为阴性对照,MTS法检测细胞增殖能力,流式细胞仪检测细胞周期和凋亡变化;Western blot检测CDCA3、CyclinD1和cleaved caspase-3蛋白的表达变化。结果食管癌组织中CDCA3 mRNA的表达高于癌旁组织(P<0.05),Eca109、Kyse-170和TE1细胞中CDCA3 mRNA表达高于人食管鳞状上皮细胞(P<0.05)。沉默CDCA3表达后,Eca109细胞和TE1细胞的增殖率降低,G0/G1期细胞比例增加(P<0.05)。沉默CDCA3表达后,Eca109细胞和TE1细胞的凋亡率增加(P<0.05)。沉默CDCA3表达可降低CyclinD1蛋白表达和增加cleaved caspase-3蛋白表达(P<0.05)。结论CDCA3 mRNA在食管癌组织和细胞系中均高表达,干扰CDCA3的表达可能通过降低CyclinD1蛋白表达抑制细胞增殖,增加cleaved caspase-3蛋白表达促进细胞凋亡,提示CDCA3可能为食管癌的潜在治疗靶标。     

RNA干扰质粒对胰腺癌细胞系Panc-1原癌基因AKT2表达的影响

  目的  探讨RNA干扰质粒抑制胰腺癌细胞系Panc-1原癌基因AKT2的表达对胰腺癌细胞生长和凋亡的影响, 并初步探讨其作用机制。  方法  选择胰腺癌细胞系Panc-1, 构建特异性抑制AKT2表达的RNA干扰质粒, 瞬时和稳定转染胰腺癌细胞, 采用MTT法及软琼脂克隆形成实验检测胰腺癌细胞生长能力, Heochst染色及Annexin V-FITC/PI染色法检测细胞凋亡情况, 通过Western blot方法检测凋亡蛋白caspase-3表达; 并进行裸鼠移植瘤体内转染实验。  结果  采用RNA干扰质粒沉默胰腺癌细胞系Panc-1原癌基因AKT2, 能够有效抑制胰腺癌细胞Panc-1体外生长能力、促进细胞凋亡, 诱导凋亡激酶caspase-3的表达; 动物体内实验结果显示, 干扰质粒能够有效抑制胰腺癌细胞系Panc-1在动物体内的成瘤能力。  结论  RNA干扰质粒抑制原癌基因AKT2表达, 可有效抑制胰腺癌细胞生长, 促进凋亡, 针对原癌基因AKT2的基因治疗对胰腺癌具有重要的潜在应用价值。     

The prognosis of patients with dissociated virological and immunological responses to HAART

Background

While HAART allows for the reconstitution of immune functions in most treated HIV patients, failure to achieve a significant increase in circulating CD4+ T cells despite undetectable viremia occurs.

Methods

A retrospective study was conducted to evaluate the treatment outcome in a subgroup of 232 patients who after 3.1 years of treatment had not achieved desirable immune reconstitution despite a good virological response to HAART.

Results

After a further 3.6 ± 2.4 years of HAART, 82 (35.3%) patients achieved immune reconstitution (565.2 ± 174.6 CD4 cells/μl), while 149 (64.2%) patients did not (268.8 ± 91.1 cells/μl); the difference in the achieved CD4 counts between these subgroups was significant (P < 0.01). One patient experienced treatment failure. Eleven patients died to the end of follow-up, of which 10 with a continuously dissociated response. Factors associated with immune recovery included clinical AIDS at HAART initiation (OR: 0.4, 95% CI: 0.24–0.81, P < 0.01), usage of PIs and of drugs from all three classes (OR: 1.7, 95% CI: 1.0–3.0, P = 0.046 and OR: 4.5, 95% CI: 1.15–18.19, P = 0.03, respectively), and a rise in CD4 count to over 200 cells/μl after the first 3.1 years of treatment (OR: 5.3 95% CI: 2.6–11.0, P < 0.01). Achievement of a rise in CD4 count to over 200 cells/μl after the first 3.1 years of treatment was an independent predictor of immune reconstitution in the following period.

Conclusion

If patients on HAART reach CD4 cell counts of above 200 cells/μl in the first 3 years, immune recovery is possible after at least 6 years of treatment.     

Kinematic and kinetic modifications in walking pattern of hip osteoarthritis patients induced by intra-articular injections of hyaluronic acid

Background

A growing body of evidence points to the efficacy of intra-articular injections of hyaluronic acid, in dealing with pain and function in hip osteoarthritis. To date, however, no data exist as to this treatment's effect on walking pattern.

Methods

We performed a prospective, open study in order to verify, in a group of 20 hip osteoarthritis patients (12 men, 8 women, mean age 60.5, range 47–73), the clinical effects of 3 intra-articular injections of 2 ml of hyaluronic acid in the hip (1/week) in terms of pain and function at 1 (T1), 3 (T2) and 6-month (T3) follow-ups, as well as changes in the kinematics and kinetics of gait at 6-month follow-up.

Findings

Pain as measured with visual analog scale significantly dropped after this procedure (P < 0.0001). A significant improvement was noted regarding stiffness (P = 0.005) and disability (P = 0.04), as measured by the Western Ontario and McMaster Universities osteoarthritis index. As regards gait analysis, patients at T3 walked with higher cadence (P = 0.004) and stride length (P = 0.02) compared to T0. Moreover, a significant increase for the pelvic tilt at heel contact (P = 0.0004) and for hip flexion–extension moment at loading response sub-phases of gait cycle (P = 0.02) was noted at T3.

Interpretation

In line with current literature, our patients display clinical improvement 6 months after intra-articular injections of hyaluronic acid, accompanied by changes in walking pattern, as measured by instrumental gait analysis. The kinematic and kinetic changes observed may be the consequence of the therapeutic effect of intra-articular injections of hyaluronic acid.     

沉默Survivin基因对鼻咽癌CNE1细胞增殖和凋亡的研究

目的：探讨 Survivin表达降低时,对鼻咽癌细胞的增殖及凋亡的影响。方法运用 RNA 干扰技术,结合 RT-PCR,Western blot技术,观察干扰 Survivin效果。干扰 Survivn后用 MTT法和 TUNEL法检测鼻咽癌细胞增殖,凋亡的情况。用 Western-blot方法检测凋亡相关蛋白 PARP,Bcl-2和Bax的表达。结果 RT-PCR检测显示 Survivin-siRNA干扰组,Survivin mRNA表达明显下调,其相对表达量为0.26±0.02,抑制率为43.7％,与对照组相比差异具有统计学意义（P＜0.05）;Western blot检测显示 Survivin-siRNA组蛋白表达下调,其蛋白相对表达量下降了57％,与对照组相比差异具有统计学意义（P＜0.05）;MTT检测显示 Survivin-siRNA组对细胞增殖有明显抑制作用,在24,48和72 h的抑制率分别为21.9％,37.1％和29.6％,与对照组相比差异具有统计学意义（P＜0.05）;Tunel染色结果显示 Survivin-siRNA组细胞凋亡数明显增高。Western blot结果显示Survivin-siRNA组凋亡蛋白PARP（89KD）显著升高,为对照组的3.9倍,抑制凋亡蛋白Bcl-2降低,降低了70％,促进凋亡蛋白Bax升高,为对照组的2.4倍;同时也抑制AKT的磷酸化,磷酸化水平降低了57％,与对照组相比以上结果差异均具有统计学意义（P＜0.05）。结论干扰 Survivin表达,抑制鼻咽癌细胞增殖,促进细胞凋亡。Survivin基因可成为鼻咽癌基因治疗的一个潜在靶点。     

Effects of a histone deacetylase inhibitor, sodium butyrate, on 53-kDa protein expression and sensitivity to anticancer drugs of pancreatic cancer cells

Background: Several tumor-suppressor genes, such as 53-kDa protein (p53), are inactivated in some pancreatic cancers. The lack of a functional p53 has been proposed to be a component of resistance to chemotherapy, resulting in the inhibition of apoptosis. Therefore, reintroduction of wild-type p53 is a commonly used gene therapy strategy for the treatment of various types of cancer, including pancreatic cancer.Objective: The aim of this study was to examine the ability of the histone deacetylase inhibitor, sodium butyrate (NaB), to modulate the expression of p53.Methods: Five human pancreatic carcinoma cell lines (SW-1990, BxPC-3, PANC-1, MIA PaCa-2, JHP-1) were utilized. Two of the cell lines (SW-1990 and JHP-1) lacked p53 expression, as determined by Western blot analysis, and were investigated further. Expression of p53 was determined by densitometry of all bands present in the Western blot. Drug sensitivity was measured with a tetrazolium-based assay by exposing the cells to graded concentrations of NaB and/or anticancer drugs (cisplatin, fluorouracil, SN-38, and paclitaxel). Apoptosis was observed using gel electrophoresis.Results: In the SW-1990 and JHP-1 cell lines, use of 1 mM NaB was found to induce histone acetylation and p53 expression compared with those not treated with NaB (P = 0.01 and P = 0.018, respectively). Sensitivity to cisplatin (P = 0.021), fluorouracil (P = 0.046), and SN-38 (P = 0.039) was significantly enhanced by NaB treatment compared with nontreatment. However, sensitivity to paclitaxel was not significantly different between untreated and NaB-treated cells. A higher frequency of apoptosis was observed in NaB-treated cells compared with that of control cells.Conclusion: This in vitro study found that NaB induced p53 expression in 2 pancreatic cancer cell lines (SW-1990 and JHP-1). Moreover, NaB acted on a biochemical modulator for antieuplastic therapy. Future research is necessary to assess the value of these findings.     

The impact of chest compression rates on quality of chest compressions - a manikin study

Purpose

Chest compressions are often performed at a variable rate during cardiopulmonary resuscitation (CPR). The effect of compression rate on other chest compression quality variables (compression depth, duty-cycle, leaning, performance decay over time) is unknown. This randomised controlled cross-over manikin study examined the effect of different compression rates on the other chest compression quality variables.

Methods

Twenty healthcare professionals performed 2 min of continuous compressions on an instrumented manikin at rates of 80, 100, 120, 140 and 160 min⁻¹ in a random order. An electronic metronome was used to guide compression rate. Compression data were analysed by repeated measures ANOVA and are presented as mean (SD). Non-parametric data was analysed by Friedman test.

Results

At faster compression rates there were significant improvements in the number of compressions delivered (160(2) at 80 min⁻¹ vs. 312(13) compressions at 160 min⁻¹, P < 0.001); and compression duty-cycle (43(6)% at 80 min⁻¹ vs. 50(7)% at 160 min⁻¹, P < 0.001). This was at the cost of a significant reduction in compression depth (39.5(10) mm at 80 min⁻¹ vs. 34.5(11) mm at 160 min⁻¹, P < 0.001); and earlier decay in compression quality (median decay point 120 s at 80 min⁻¹ vs. 40 s at 160 min⁻¹, P < 0.001). Additionally not all participants achieved the target rate (100% at 80 min⁻¹ vs. 70% at 160 min⁻¹). Rates above 120 min⁻¹ had the greatest impact on reducing chest compression quality.

Conclusions

For Guidelines 2005 trained rescuers, a chest compression rate of 100–120 min⁻¹ for 2 min is feasible whilst maintaining adequate chest compression quality in terms of depth, duty-cycle, leaning, and decay in compression performance. Further studies are needed to assess the impact of the Guidelines 2010 recommendation for deeper and faster chest compressions.     

Greater Q angle may not be a risk factor of patellofemoral pain syndrome

Background

A greater Q-angle has been suggested as a risk factor for Patellofemoral Pain Syndrome. Greater frontal plane knee moment and impulse have been found to play a functional role in the onset of Patellofemoral Pain Syndrome in a running population. Therefore, the purpose of this investigation was to determine the relationship between Q-angle and the magnitude of knee abduction moment and impulse during running.

Methods

Q-angle was statically measured, using a goniometer from three markers on the anterior superior iliac spine, the midpoint of the patella and the tibial tuberosity. Thirty-one recreational runners (21 males and 10 females) performed 8–10 trials running at 4 m/s (SD 0.2) on a 30 m-runway. Absolute and normalized knee moment and impulse were calculated and correlated with Q-angle.

Findings

Negative correlations between Q-angle and the magnitude of peak knee abduction moment (R² = 0.2444, R = − 0.4944, P = 0.005) and impulse (R² = 0.2563, R = − 0.5063, P = 0.004) were found. Additionally, negative correlations between Q-angle and the magnitude of weight normalized knee abduction moment (R² = 0.1842, R = − 0.4292, P = 0.016) and impulse (R² = 0.2304, R = − 0.4801, P = 0.006) were found.

Interpretation

The findings indicate that greater Q-angle, which is actually associated with decreased frontal plane knee abduction moment and impulse during running, may not be a risk factor of Patellofemoral Pain Syndrome.     

The angiotensin receptor blocker, telmisartan, reduces and stabilizes atherosclerosis in ApoE and AT1aR double deficient mice

The renin-angiotensin system (RAS) plays critical roles in the pathogenesis of atherosclerosis. Clinical studies demonstrate that pharmacological blockade of RAS with Angiotensin II type 1 receptor (AT1R) blockers (ARBs) is effective in the treatment of patients with cardiovascular diseases. Recent studies reported that telmisartan, an ARB, has a partial agonistic effect on peroxisome proliferator-activated receptor-gamma (PPAR-γ). The role of PPAR-γ-mediated signaling has been implicated in regulation of not only metabolic disorders but also atherosclerosis. Here, we investigated the effects of telmisartan, which is not related to AT1R blockade, using AT1aR and apolipoprotein E (ApoE) double-deficient (ApoE−/−AT1R−/−) mice in vivo. Both genetic ablation of AT1R in ApoE-deficient (ApoE−/−) mice and administration of telmisartan (10 mg/kg/day) to ApoE−/− mice for 20 weeks reduced the development of atherosclerosis (P < 0.05, respectively). Telmisartan decreased lipid deposition (P < 0.01) and increased collagen contents (P < 0.05) in plaques in ApoE−/− mice. Administration of telmisartan to ApoE−/−AT1aR−/− mice also inhibited the progression of atherosclerosis in aorta (P < 0.05) even in mice, which have no AT1aR genetically. Moreover, in these mice, telmisartan decreased macrophage accumulation and lipid deposition, and increased collagen contents in plaques in aortic root (P < 0.05, respectively), indicating stabilization of plaques. Telmisartan-treated ApoE−/−AT1aR−/− mice showed lower body weight and higher plasma high-density lipoprotein levels compared with vehicle-treated mice (P < 0.05, respectively). Telmisartan lowered systolic and diastolic blood pressure in ApoE−/−AT1aR−/− mice (P < 0.01). These results suggest that telmisartan has protective effects on the development of atherosclerosis and metabolic disorders beyond AT1R blockade in ApoE-deficient mice.     

Relationships between clinical measures of static foot posture and plantar pressure during static standing and walking

Background

Information is limited about the relationships between clinical measures of static foot posture and peak plantar pressures under the medial column of the foot. The purpose was to examine these relationships during static standing and walking.

Methods

A single-group exploratory design using correlation and regression was used to determine relationships. Ninety-two healthy volunteers participated. Clinical measures of static foot posture including arch index, navicular drop and navicular drift were obtained during static standing. Peak plantar pressures under the hallux, medial forefoot, medial midfoot, and medial rearfoot were obtained during standing and walking.

Findings

Static foot posture was related to peak plantar pressures during standing and walking, but the strengths of relationships ranged from poor to fair. During standing, navicular drop was correlated (P ≤ 0.05) with hallux (r = 0.29) and medial forefoot (r = − 0.17) pressures, while arch index (r = − 0.17) and navicular drift (r = 0.25) were correlated (P ≤ 0.05) with hallux pressure. During walking, arch index, navicular drop and navicular drift were correlated (P ≤ 0.05) with hallux and medial forefoot pressures (r range − 0.30 to 0.41), while arch index (r = − 0.15) and navicular drop (r = 0.16) were correlated (P ≤ 0.05) with medial midfoot pressure. Regression models predicted (P ≤ 0.05) hallux (R² = 0.08) and medial midfoot (R² = 0.05) pressures during standing, and hallux (R² = 0.18), medial forefoot (R² = 0.07), and medial rearfoot (R² = 0.05) pressures during walking.

Interpretation

In healthy participants, lower arch foot postures are associated with greater pressures under the hallux and medial mid-foot and lower pressures under the medial forefoot, but the strength of these relationships may be only poor to fair.     

Changes in infraspinatus cross-sectional area and shoulder range of motion with repetitive eccentric external rotator contraction

Background

Repetitive eccentric loading results in muscle damage and subsequent changes in muscle stiffness and edema accumulation, which manifest as reduced joint range of motion and increased muscle cross-sectional area. The purpose of the study was to evaluate changes in shoulder range of motion and the infraspinatus cross-sectional area with repetitive eccentric contraction.

Methods

Twenty physically active participants performed 9 sets of 25 repetitions of eccentric external rotator contractions. The ultrasonographic measurement of the infraspinatus cross-sectional area, and shoulder internal/external rotation and horizontal adduction range of motion were measured before, immediately after, and 24 h after the intervention.

Findings

Infraspinatus cross-sectional area significantly increased from baseline immediately after exercise (P < 0.001), and remained elevated from baseline at the 24-hour follow up (P < 0.001). Internal rotation and horizontal adduction range of motion did not change significantly between baseline and post-exercise (P > 0.05), but were significantly decreased at the 24-hour follow up from the baseline (internal rotation: P < 0.001, horizontal adduction: P < 0.001) and the immediate post-exercise (internal rotation: P = 1.012, horizontal adduction: P = 0.016).

Interpretation

These changes observed after the eccentric contractions may have implications for injury development in pitchers, because 1) the infraspinatus endures repetitive eccentric loading with pitching and 2) decreased internal rotation and horizontal adduction range of motion have been linked to upper extremity injuries. However, since the muscle response after eccentric loading varies by the task and previous exposure to similar stress, future study needs to investigate the time course of recovery of the muscle cross-sectional area and range of motion after pitching in competitive pitchers.     

Embryonic develop-associated gene 1 is overexpressed and acts as a tumor promoter in thyroid carcinoma

Embryonic develop-associated gene 1 (EDAG-1), a hematopoietic tissue-specific protein, is usually highly expressed in the placenta, fetal liver, bone marrow and leukemia cells, but the expression status in normal or solid tumor tissues is rarely reported. In this study, we found that EDAG-1 was up-regulated in thyroid carcinoma tissues and cells. Knockdown of EDAG-1 suppressed proliferation and enhanced cisplatin-induced apoptosis of thyroid carcinoma cells. We also demonstrated that knockdown of EDAG-1 inactivated the phosphatidylinositol-3 kinase (PI3K)/Akt signaling pathway in vitro and in vivo. Moreover, knockdown of EDAG-1 suppressed tumorigenesis of thyroid carcinoma in vivo. Taken together, these results suggest that EDAG-1 regulates the proliferation and apoptosis of thyroid carcinoma via the PI3K/Akt signaling pathway.     

Lack of effect of moderate-duration static stretching on plantar flexor force production and series compliance

Background

The effects of an acute bout of moderate-duration static stretching on plantar flexor force production, series compliance of the muscle–tendon unit, and levels of neuromuscular activation were examined.

Methods

Eighteen active individuals (9 men and 9 women) performed four 45-s static plantar flexor stretches and a time-matched control of no stretch (where subjects remained seated in the dynamometer for 4 min with no stretch being performed). Measures of peak isometric moment, rate of force development, neuromuscular activation (interpolated twitch technique and electromyography), twitch force characteristics, passive moment during stretch, and tendon elongation during maximal voluntary contractions were taken before and after the stretching.

Findings

Despite a significant stress–relaxation response during stretch (9.3%, P < 0.01) there were no significant differences in peak isometric moment (P = 0.35; effect size 0.13), rate of force development (P = 0.93; effect size 0.01), neuromuscular activation (interpolated twitch: P = 0.86; electromyography: P = 0.09; effect size 0.02), or tendon elongation (P = 0.61; effect size 0.07) after stretching. Twitch characteristics were also unchanged after stretching, although there was a reduction in the rate of twitch torque relaxation (RR_t; P < 0.01).

Interpretation

The acute bout of moderate-duration static stretching did not impair the force generating capacity of the plantar flexors or negatively affect muscle–tendon mechanical properties. Static stretching may not always have detrimental consequences for force production. Thus, clinicians may be able to apply moderate-duration stretches to patients without risk of reducing muscular performance.     

Biomechanical changes accompanying unilateral and bilateral use of laterally wedged insoles with medial arch supports in patients with medial knee osteoarthritis

Background

Laterally wedged insoles have controversial effect in treating medial compartment knee osteoarthritis. This study examined the effects of unilateral and bilateral use of insoles having medial arch supports and of different inclinations on the frontal plane external hip, knee, subtalar moments and pelvic alignment.

Methods

Kinetic and kinematic gait parameters were collected from 21 patients with primary medial knee osteoarthritis. The insoles' inclinations were 0, 6 and 11°, where each of the 6° and 11° was used once unilaterally and another bilaterally while the 0° was used bilaterally as a control.

Findings

The Multivariate Analysis of Variance revealed significant increase in the external subtalar eversion moment using either of the 6° or 11° laterally wedged vs the 0° non-wedged insole conditions (P = 0.003). Moreover, there were significant increases in the external eversion moment using the 11° vs the 6° insole conditions (P < 0.05). However, there were no significant differences for the remaining tested variables (P > 0.05). The bivariate correlations revealed significant negative correlations between the subtalar eversion and knee adduction moments (r = − 0.409, P = 0.000) and the subtalar eversion and hip adduction moments (r = − 0.226, P = 0.049), and positive correlation between the hip and knee adduction moments (r = 0.268, P = 0.019).

Interpretation

The non-significant reduction in the external knee adduction moment may question the efficacy of using wedged insoles having medial arch supports in treating patients with medial knee osteoarthritis. Additionally, using such insoles did not produce appreciable mechanical effects on remote articulations as the hip and pelvis.     

Gallic Acid as a cancer-selective agent induces apoptosis in pancreatic cancer cells

Background: Gallic acid (GA) is a plant phenol isolated from water caltrop which is reported to have anti-inflammatory and anti-cancer effects. In this study, the antiproliferative effect of GA on human pancreatic cancer cell lines CFPAC-1 and MiaPaCa-2 as well as hepatocytes HL-7702 as normal cells was examined. Particularly, the mechanism of GA-induced apoptosis in MiaPaCa-2 cells in vitro was further studied. Methods: Cell viability was measured using SRB assay, and apoptosis was detected by Hoechst staining and annexin V-PI staining assays. Mitochondrial membrane potential was detected by rhodamine-123 staining. Flow cytometry analysis was employed to detect the apoptosis-related events. Results: GA inhibited the proliferation of CFPAC-1 and MiaPaCa-2 cells in a time- and dose-dependent manner, with IC(50)S of 102.3 ± 2.4 and 135.2 ± 0.6 μM at 48 h, respectively. GA treatment led to the increased proportion of cell apoptosis from 12.5 ± 0.72 to 78.3 ± 2.48% at the concentrations of 6.25 and 25.0 μg/ml, which was evidenced again by chromatins staining assay. Also, GA activated caspase-3, caspase-9, and reactive oxygen species, elevated Bax expression and [Ca(2+)](i) and reduced mitochondrial membrane potential (ΔΨm) in MiaPaCa-2 cells. Remarkably, when compared with human normal cells HL-7702 (IC(50) >100 μg/ml), GA showed selective toxicity for cancer cells. Conclusions: GA can function as a cancer-selective agent by inducing apoptosis in MiaPaCa-2 cells via the mitochondria-mediated pathways. To the best of our knowledge, GA should open up new opportunities for the therapy of pancreatic cancer.     

Molecular mechanisms of therapeutic hypothermia on neurological function in a swine model of cardiopulmonary resuscitation

Objective

To explore the molecular mechanisms by which mild hypothermia following resuscitation improves neurological function in a porcine model of cardiac arrest.

Methods

Thirty-three inbred Chinese Wuzhishan (WZS) minipigs were used. After 8 min of untreated ventricular fibrillation (VF), the surviving animals (n = 29) were randomly divided into two groups including serum group (n = 16) and molecular group (n = 13). Serum group animals were used to measure porcine-specific tumour necrosis factor-alpha (TNF-α), interleukin (IL-6, IL-10), matrix metalloproteinase (MMP9), Aquaporin-4 (AQP4), tissue inhibitor to metalloproteinase-1 (TIMP1), neuron-specific enolase (NSE) and S100B at 0.5 h, 6 h, 12 h, 24 h and 72 h recovery by enzyme-linked immunosorbent assay (ELISA). Molecular group animals were used to measure cerebral cortex messenger RNA (mRNA) and protein expression of nuclear factor-κB (NF-κB), MMP9 and AQP4 by real-time (RT) quantitative polymerase chain reaction (PCR) and Western blotting at 24 h and 72 h recovery. Animals were further divided into either normothermia or hypothermia groups. Hypothermia (33 °C) was maintained for 12 h using an endovascular cooling device. Swine neurologic deficit scores (NDS) were used to evaluate neurological function at 24-h and 72-h recovery.

Results

Twenty-nine of the 33 (87.9%) animals were successfully resuscitated. The hypothermia group exhibited higher survival rates at 24 h (75%) and 72 h (62.5%) compared to the normothermia group (37.5% and 25%, respectively). Hypothermia markedly inhibited expression of NF-κB, TNF-α, MMP9 and NSE, and promoted expression of TIMP1 (P < 0.01). The mean NDS at 24-h and 72-h recovery was 112.5 and 61, respectively, in the hypothermic group, and 230 and 207.5, respectively, in the normothermia group.

Conclusion

Brain protection induced by hypothermia involves inhibition of inflammatory and brain edema pathways.     

Effects of continuous haemofiltration on serum enzyme concentrations, endotoxemia, homeostasis and survival in dogs with severe heat stroke

Aim

To examine the effectiveness of continuous haemofiltration as a treatment for severe heat stroke in dogs.

Methods

Dogs were randomly allocated to a control or continuous haemofiltration group (both n = 8). Heat stroke was induced by placing anaesthetised dogs in a high temperature cabin simulator. Upon confirmation of heat stroke (rectal temperature > 42 °C, mean arterial pressure (MAP) decrease > 25 mmHg), dogs were removed from the chamber and continuous haemofiltration was initiated and continued for 3 h for dogs in the continuous haemofiltration group. Dogs in the control group were observed at room temperature.

Results

Rectal temperature, haemodynamics, pH, blood gases and electrolyte concentrations rapidly returned to baseline in the continuous haemofiltration group, but not the control group. After 3 h, rectal temperature was 36.68 ± 0.51 °C in the continuous haemofiltration group and 39.83 ± 1.10 °C in the control group (P < 0.05). Continuous haemofiltration prevented endotoxin and all serum enzyme concentrations from increasing and caused malondialdehyde concentrations to decrease. After 3 h, endotoxin concentrations were 0.14 ± 0.02 EU ml⁻¹ in the continuous haemofiltration group and 0.23 ± 0.05 EU ml⁻¹ in the control group (P = 0.003), while malondialdehyde concentrations were 4.86 ± 0.61 mmol l⁻¹ in the continuous haemofiltration group and 8.63 ± 0.66 mmol l⁻¹ in the control group (P < 0.001). Five dogs died in the control group within 3 h, whereas no dogs died in the continuous haemofiltration group.

Conclusions

Continuous haemofiltration rapidly reduced body temperature, normalised haemodynamics and electrolytes, improved serum enzyme concentrations and increased survival in dogs with heat stroke. Continuous haemofiltration may be an effective treatment for heat stroke.     

Gender and fatigue have influence on knee joint control strategies during landing

Background

Gender and fatigue are thought to affect the anterior cruciate ligament injury risk. In spite of much effort, the influence of these factors on knee joint biomechanics is still under discussion. The purpose of this study was to investigate kinematics, kinetics, and active muscle control strategies of the knee joint across gender in fatigue conditions during a landing task.

Methods

Thirteen females and thirteen males performed two-legged landings before and after a closed kinetic chain exercise protocol. Knee joint kinematics and vertical ground reaction forces were assessed as well as electromyography of the quadriceps, hamstring, and gastrocnemius muscles.

Findings

Females landed with increased knee flexion velocities (P < 0.001) and knee joint abduction angles (P < 0.01). Compared to males, females also showed different muscle activation patterns such as a delayed activation of the lateral hamstring (P < 0.05) and the m. vastus lateralis (P < 0.05) during the preparatory phase of the landing. Fatigue led to a reduced pre-activation of the medial and lateral hamstrings (P < 0.05 and P < 0.001) and the gastrocnemius muscle (P < 0.05) both in males and females.

Interpretation

The gender differences in knee flexion velocity, abduction angle, and muscle activation suggest that females and males possess different neuromuscular strategies to control the knee joint during dynamic landing movements. These differences as well as decreased hamstring and gastrocnemius muscle activity, due to fatigue, provide evidence for different knee joint control in females as well as in fatigued conditions.