[The change in bone marrow after intraosseous hypertonic saline-hydroxyethyl starch infusion for resuscitation of hemorrhagic shock in dog].

OBJECTIVE: To observe the change in bone marrow after intraosseous hypertonic saline- hydroxyethyl starch (HSH) in dogs with hemorrhagic shock. METHODS: Eighteen male dogs were randomly divided into sham group, normal saline (NS) group (0.9% sodium chloride) and HSH group (7.5% sodium chloride-6% hydroxyethyl starch). Wiggers graded controlled hemorrhagic shock model was reproduced in NS and HSH groups, and above solutions were injected into bone marrow of the tibia in low dosage (4 ml/kg) rapidly (within 5 minutes). The sham group did not lose blood or receive infusion. CD34(+) expression of peripheral blood and bone marrow, and the change in picture of different hematopoietic series in bone marrow were examined before and after bone marrow resuscitation. Pathological change in bone marrow was examined after different resuscitation fluid infusion. RESULTS: After 48 hours of resuscitation, the percentage of CD34(+) in bone marrow and peripheral blood in HSH group were significantly higher than that of NS group [peripheral blood: (6.915+/-1.178)% vs. (4.848+/-0.527)%, bone marrow: (7.900+/-0.648)% vs. (6.875+/- 0.403)% , both P<0.05]. One week after resuscitation, the erythrocyte count and granulocytic count in NS group were significantly increased compared with sham group (erythrocyte count: 0.289+/-0.016 vs. 0.253+/-0.014, granulocytic count: 0.615+/-0.019 vs. 0.560+/-0.013, both P<0.05], lymphocytic count was significantly decreased (0.049+/-0.007 vs. 0.132+/-0.015, P<0.05). In the HSH group, erythrocyte count (0.273+/-0.012), lymphocytic count (0.162+/-0.014) were all significantly increased (both P<0.05), while granulocytic count (0.517+/-0.038) was significantly decreased (P<0.05). After 4 week of resuscitation, the change in hemopoiesis series in NS group and HSH group recovered to normal level. The bone marrow morphology changed slightly, and no bone necrosis occurred. CONCLUSION: HSH in small amount through intraosseous space is safe and effective as a fluid resuscitation measure for shock, and little change in bone marrow has been found after the infusion.

[Effects of astragalus injection on extracellular signal-regulated kinase pathway in cultured normal cardiac myocytes].

OBJECTIVE: To investigate the dose-effect relationship and time-effect relationship of astragalus injection on the protein expression of nitric-oxide synthase (NOS) and extracellular signal-regulated kinase (ERK) in cultured cardiac myocytes. METHODS: (1) Ventricular myocytes of Sprague-Dawley rats were isolated, cultured, and incubated with astragalus injection of the concentrations of 0, 20, 100, 200, or 400 ml/L for 8 days. Cell growth curve was drawn. Automatic biochemical analyzer was used to detect the level of creatine kinase (CK). (2) Western blotting was used 2 h after incubation to observe the protein expression of ERK and nitric oxide synthase (NOS). (3) Myocytes were incubated with astragalus injection (200 ml/L) or with astragalus and PD98059, an ERK inhibitor, for 5, 15, 30, 60, and 120 min respectively, and Western blotting was used to detect the protein expression of ERK and NOS. RESULTS: Astragalus of different concentrations showed no effect on the cell proliferation. Only the CK level of the astragalus 200 ml/L group was significantly higher than that of the control group (P < 0.05). Astragalus showed no dose-dependent effect on the ERK expression, and dose-dependently reduced the NOS expression level. 15 min after the co-incubation with astragalus 200 ml/L the ERK level of the myocytes decreased transiently, and then began to increase gradually since 15 min after the co-incubation; the NOS level began to increase since 60 min after the co-incubation (P < 0.05). After the addition of PD98059 the ERK expression significantly decreased (P < 0.05). CONCLUSION: Astragalus injection has modulation effect on ERK and NOS expression. This effect is enhanced time-dependently at appropriate Astragalus concentration, but attenuated by high dose Astragalus injection.

Evaluation of 29 indicators for the prognosis of advanced non-small cell lung cancer with cytokine-induced killer cell therapy combined with chemotherapy.

The aim of the present study was to evaluate 29 whole blood or serum indicators to identify factors able to predict clinical outcome following cytokine-induced killer (CIK) cell therapy combined with chemotherapy in patients with advanced non-small cell lung cancer (NSCLC), and to evaluate the 5-year prognosis of the patients. From March 2008 to October 2013, 42 patients with advanced NSCLC (stages III and IV) were enrolled in the study. These patients were from a single hospital, and had been treated with CIK therapy combined with chemotherapy. Evaluation of the correlation between prognosis and age, gender, tumor stage, surgery resection status, number of CIK therapy cycles, tumor subtype, and the differential whole blood or serum indicators were analyzed by Kaplan-Meier methods and the log-rank test. The prognostic factors were analyzed by Cox proportional models. The median progression-free survival (mPFS) time of patients with high expression levels of albumin [20.0 months; 95% confidence interval (CI): 17.4-22.6 months] was significantly longer than the mPFS for patients with low expression levels of albumin (36.0 months; 95% CI: 24.7-47.3 months) (P=0.034). Other factors demonstrated no significant difference. Following analysis using the Cox proportional hazards regression model, the number of CIK therapy cycles (P=0.041) and the expression level of albumin (P=0.038) were revealed to be independent prognostic factors following the use of CIK cell therapy combined with chemotherapy for patients with advanced NSCLC. The risk of adverse outcomes in patients receiving ≥4 CIK therapy cycles and in patients with increased expression levels of albumin were 0.38 (95% CI: 0.14-1.13) and 0.32 (95% CI: 0.10-1.24)-fold those of patients receiving <4 CIK therapy cycles and with decreased expression levels of albumin, respectively. The serum albumin concentration may therefore be a predictor of the 5-year survival rate of patients with advanced NSCLC treated with CIK cell therapy combined with chemotherapy; patients with high expression levels of albumin may have a better prognosis in comparison with patients with low expression levels of albumin.

Relationship of serum polyunsaturated fatty acids with cytokines in colorectal cancer.

AIM: To investigate the relationship of serum levels of polyunsaturated fatty acid (PUFA) with kinds of cytokines in colorectal cancer (CRC). METHODS: Serum samples of 100 CRC patients were collected. The concentration of total n-3 PUFA which included C18:3 n-3, C20:5 n-3, C22:5 n-3, C22:6 n-3 and the total n-6 PUFA included C18:2 n-6, C18:3 n-6, C20:3 n-6, C20:4 n-6, and C22:5 n-6 were detected on GC-2010 Plus Gas Chromatograph with a OmegawaxTM 250 column. Cytokines were detected by MagPlexTM-C microspheres. P values for the trend were estimated by creating a continuous variable using the median value within quartiles. RESULTS: Interleukin-6 (IL-6) showed significantly positive association with the C20:4 n-6 (P for trend = 0.004). Interferon gamma (IFN-γ) showed significant positive association with the C22:5 n-3 (P for trend = 0.035). IL-8 and matrix metalloproteinase-9 (MMP-9) showed significant inverse association with the C22:6 n-3 (P for trend = 0.049, and 0.021). MMP-2 showed significant inverse association with the C20:5 n-3 (P for trend = 0.008). MMP-7 showed significantly positive association with the ratio of n-6 PUFA and n-3 PUFA (P for trend = 0.008). MMP-7 also showed significantly inverse association with the ratio of C20:4 n-6 and (n-6 PUFA + n-3 PUFA) (P for trend = 0.024). IL-10 (P for trend = 0.023) and IL-6 (P for trend = 0.036) showed significantly positive association with the ratio of C20:4 n-6 and C20:5 n-3. CONCLUSION: Our data suggested that serum levels of PUFA is related to the inflammation of CRC, and also play different role in regulation of immune response.

[Modeling of acute respiratory distress syndrome in canine after inhalation of perfluoroisobutylene and preliminary study on mechanisms of injury].

OBJECTIVE: To establish of acute respiratory distress syndrome (ARDS) model in canine after inhalation of perfluoroisobutylene (PFIB), and to observe the progressing of lung injury, and to study the mechanisms of injury. METHODS: A device of inhalation of PFIB for canine was made. The concentration of PFIB was 0.30 - 0.32 mg/L. Serum IL-6 and IL-8 were dynamically measured. Clinical manifestations, pathology of organs in canine were observed. RESULTS: (1) During inhalation, the concentration of PFIB remained stable; (2) After inhalation, blood arterial oxygen partial pressure fell gradually, and eventually met the criteria for diagnosing ARDS; (3) The level of IL-8 in serum rises significantly after inhalation (P < 0.05), whereas that of IL-6 was not obviously altered (P > 0.05); (4) Within 6 hours after inhalation, no abnormality in canine was observed, but afterwards symptoms gradually appeared, and typical breath of ARDS, such as high frequency and lower level could be seen in later phase; (5) Pathological examination showed severe congestion, edema and atelectasis in most part of both lungs, and signs of anoxia in other organs. CONCLUSIONS: (1) The device designed is capable of ensuring control of inhalation of PFIB; (2) Exposure to PFIB for 30 mins, canines all met the criteria for diagnosing ARDS 22 hours after inhalation, therefore the modeling is successful; (3) PFIB specifically damages the lung by causing excessive inflammation.

Clinical study of autologous cytokine-induced killer cells for the treatment of elderly patients with diffuse large B-cell lymphoma.

To evaluate the effectiveness and safety of autologous cytokine-induced killer (CIK) cells in elderly patients with diffuse large B-cell lymphoma. Peripheral blood mononuclear cells (PBMC) were isolated from nine elderly patients with diffuse large B-cell lymphoma. PBMCs were augmented by priming with interferon gamma (IFN-γ) followed by IL-2 and monoclonal antibody (mAb) against CD3. Autologous CIK cells (range 5 × 10(9)-1 × 10(10)) were then infused back to individual patients; infusion was repeated every 4 weeks for 32 weeks (eight cycles). Patients were assessed for changes in lymphocyte subgroup, tumor-related biological parameters, imaging characteristics, the condition of remission, quality of life (QOL), and survival. Prior to CIK infusion, two patients were in complete remission and seven patients were in partial remission. After autologous CIK cell transfusions, the proportion of CD3+, CD3+CD8+, and CD3+CD56+ cells were significantly increased compared with baseline (P < 0.05); whereas serum levels of ß2-microglobulin and LDH were significantly decreased (P < 0.05). The lymphoma symptoms were reduced and QOL was improved (P < 0.05) in all patients. All patients achieved complete remission at study endpoint. No adverse reactions were reported. Autologous CIK cell immunotherapy is safe and efficacious for the treatment of elderly patients with diffuse large B-cell lymphoma.