BACKGROUND: Allogeneic blood transfusion in humans of the same blood type has been implemented, but studies regarding swine blood type and how to perform allogeneic blood transfusion have been rarely reported. OBJECTIVE: To investigate the feasibility of preoperative autologous blood donation using the leap-frog technique for cardiopulmonary bypass surgery in a swine model.METHODS: Sixteen domestic swine were randomly divided into two groups: autologous blood donation and allogeneic blood transfusion. Another four swine were used as blood donors. Two groups of swine underwent cardiopulmonary bypass surgery. The autologous blood donation group received self-transfusion reserved before surgery while the allogeneic blood transfusion group received the same amount of allogeneic blood. Hemoglobin (Hb) concentration and hematocrit (Hct) level prior to and after donation in the autologous blood donation group was recorded. Hb concentration in the two groups was recorded prior to, during, immediately after, and 1 day after the surgery. RESULTS AND CONCLUSION: The total blood volume of each experimental swine was (2500±428) mL. For the autologous blood donation group, the predicted blood volume of the first donation was (501±86) mL and the actual blood volume was (493±93) mL; in the second donation, the predicted blood volume was (750±128) mL and the actual blood volume was (719±98) mL. There was a significant difference in Hb concentration and Hct level between prior to and after donation in the autologous blood donation group (P < 0.01). Hb concentration at 1 day after the surgery was significantly higher in the autologous blood donation group than in the allogeneic blood transfusion group (P < 0.01), while no significant difference in Hb concentration existed between these two groups prior to, during, and immediately after the surgery (P > 0.05). Compared with prior to surgery, Hb concentration in each group was significantly lower at 1 day after the surgery (P < 0.01). The autologous blood donation group exhibited significantly higher survival rate of swine than the allogeneic blood transfusion group (P < 0.01). These findings indicate that compared with allogeneic blood transfusion, preoperative autologous blood donation using the leap-frog technique appears to be a safe, effective method with a high survival rate for cardiopulmonary bypass surgery.

Aim Toinvestigatewhetherexposureto Sanguinarine (SAN ) can inhibit cell proliferation in human mammary adenocarcinoma cells (MCF-7 ) and thepossiblemechanism.Methods WeexposedMCF-7 to anticancer compound SAN,cell viability was as-sessed by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT ) reduction assay. ROS was measured using confocal microscopy,expres-sion of caspase-3 ,caspase-8 and caspase-9 were calcu-latedusingchemiluminescencemethod.Results SAN remarkably inhibited growth of human mammary adeno-carcinoma MCF-7 cells by decreasing cell proliferation. ROS release and caspase-3,caspase-8,caspase-9 ex-pression were stimulated by SAN in MCF-7 ,and these changes were abolished by the antioxidant,N-acetyl-cysteine(NAC).Conclusion Regulationofcaspases expression and release from MCF-7 cells are possibly e-voked by SAN through reactive oxygen species.

Alanine Transaminase ; blood ; Hepatitis B, Chronic ; blood ; pathology ; Humans

Adult ; Antigens, CD20 ; metabolism ; Burkitt Lymphoma ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Histiocytic Necrotizing Lymphadenitis ; metabolism ; pathology ; Humans ; Lymphatic Diseases ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; Neprilysin ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Receptors, Complement 3d ; metabolism ; Submandibular Gland Diseases ; metabolism ; pathology ; Young Adult

Objective: To investigate the effect of ginsenoside-rg1 (G-Rg1) on rat’s cardiomyocytes H9c2 with its mechanism of signal pathwayin vitro.
Methods: H9c2 cells were cultured and treated in different conditions by following groups:①Blank control group,②Hypoxia alone group, the cells were treated for (2, 6, 12, 24, 48) hr respectively,③G-Rg1 group, the cells were treated by G-Rg1 at (5, 10, 50) μmol/L respectively,④YC-1 group, which is the speciifc inhibitor of hypoxia inducible factor-1α (HIF-1α),⑤YC-1 + G-Rg1 group,⑥Wortmannin group, which is the speciifc inhibitor for protein kinase B (Akt) phosphorylation and⑦Wortmannin + G-Rg1 group. Each experiment was conducted with 5 replicates. The effects of G-Rg1, hypoxia and YC-1 on cell activity and injury were studied; intracellular mRNA expressions of HIF-1α, glucose transporter-1 (GLUT-1) and heme oxygenase-1 (HO-1) were examined by RT-PCR; protein expressions of HIF-1α, GLUT-1, HO-1, activating transcription factor-6 (ATF-6), CCAAT/enhancer binding protein homologous protein (CHOP) and Akt with its signal pathway factors were measured by Western blot analysis.
Results: The time of hypoxia was negatively related to cell activity (r=-0.8580,P<0.05) and positively related to LDH overlfow rate (r=0.9201,P<0.05). G-Rg1 (10μmol/L) group showed increased cell activity than Hypoxia alone (24 hr) group (87.8% vs 62.6 %,P<0.05), while decreased LDH overlfow (25.0% vs 74.8%,P<0.05), and up-regulated mRNA expressions of HIF-1α, GLUT-1 and HO-1, P<0.05. YC-1+ G-Rg1 group had decreased cell activity than G-Rg1 group (68.0% vs 87.8%,P<0.05), while increased LDH overlfow (56.4% vs 25.0%,P<0.05). Meanwhile, YC-1 clashed the effect of G-Rg1 on protein expressions of HIF-1α, GLUT-1, HO-1, ATF-6 and CHOP,P<0.05; wortmannin clashed the effect of G-Rg1 on protein expressions of HIF-1α, CHOP,P<0.05 and suppressed the two phosphorylation sites for Akt activation,P<0.05.
Conclusion: G-Rg1 may protect rat’s H9c2 cellsin vitro by activating expressions of HIF-1α with its downstream factors and inhibiting endoplasmic reticulum stress, which might be related to the effect of G-Rg1 on Akt activation.

Objective In order to study the epidemiology of Borna disease virus (BDV) in Zunyi region and its surrounding regions in Guizhou province. Methods p24 fragment of BDV fragments in peripheral blood mononuclear cells (PBMC) from 43 patients with viral encephalitis (VE), 9 cases with multiple sclerosis (MS), 7 cases with Guillain-Barre syndrome (GBS), 5 cases with Parkinson disease (PD), 98 healthy donors and 300 goats were examined by quantitative fluorescence nested reverse transcriptase polymerase chain reaction (PCR). Gene sequence and amino acid sequence were analyzed for positive products. Results The positive rate of BDV p24 fragment in PBMC from VE (13.95 %) and MS (22.22 %) were significantly higher than in healthy donors (0 %, P<0.05). The positive rate of BDV p24 fragment in PBMC from goats was 0.67 %, without statistical difference when compared with healthy donors (P>0.05). Guillain-Barre syndrome and Parkinson disease( PD)were tested negative. The sequence of the BDV p24 fragment from the patients with VE was in conformity with that of the MS. Results presented that 3 situs consistency silent mutation when compared with strain V and its homogeneity was 96.51%. 2 situs consistency silent mutation compared with BDV/MDCK and its homogeneity was 97.67%. 2 situs consistency silent mutation when compared with C6BV and its homogeneity was 97.67 %. Sequences of the BDV p24 fragment from the goats presented 3 situs consistency silent mutation when compared with strain V and its homogeneity was 96.51%. 3 situs consistency silent mutation when compared with BDV/MDCK and its homogeneity was 96.51%. 3 situs consistency silent mutation when compared with C6BV and its homogeneity was 96.51%. However, there were no changes of encoding amino acids in all BDV p24 fragments from neuropsychiatric disorders. Conclusion Our data indicated that the BDV infection was presented in patients with VE, MS and goats from Zunyi region and its surrounding regions of Guizhou province. BDV might play a potential role in the development of VE, MS as well as having correlations with animals.

Objective: To observe the effects of electroacupuncture (EA) on the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment (CI), and to explore the mechanism of EA in improving the learning and memory abilities. Methods: A total of 100 Sprague-Dawley (SD) rats were divided into a normal group (n=10) and a model group (n=90) by random number table method. Rats in the model group were intraperitoneally injected with a small dose of streptozotocin (STZ) to establish the type 2 diabetic models, after being fed with high-fat and high-sugar diet for 1 month. Twenty CI rats were selected from the 50 successful model rats by the Morris water maze (MWM) test and randomly divided into a model group and an EA group according to the blood glucose level and MWM data (n=10). Rats in the EA group received acupuncture at Zusanli (ST 36), Neiting (ST 44) and Yishu (Extra), of which Zusanli (ST 36) and Neiting (ST 44) were stimulated by EA apparatus, 20 min/time, once a day for 6 d a week and 4 consecutive weeks. The rats in the model and the normal groups were fixed without treatment. After 4-week treatment, the random blood glucose level of the rats was measured; the learning and memory abilities of rats were measured by MWM; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptotic cells; Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex. Results: After modeling, the random blood glucose level and the escape latency tested by MWM were significantly increased, and the number of crossing the platform tested by the MWM was decreased in the EA and model groups, and were significantly different from those in the normal group (P<0.05 or P<0.01), while the differences between the model group and the EA group were not statistically significant (all P>0.05). After 4-week treatment, the random glucose level and the escape latency tested by MWM were significantly increased (both P<0.05), and the number of crossing the original platform tested by the MWM was significantly reduced (P<0.01), the protein and gene expressions of Bax and Caspase-3 were significantly increased (all P<0.001), the protein and gene expressions of Bcl-2 were significantly reduced (both P<0.001), and the number of neuron apoptosis was significantly increased (P<0.001) in the model group than in the normal group; the random blood glucose level was significantly reduced (P<0.05), the escape latency tested by MWM was significantly shortened (P<0.05), and the number of crossing the original platform tested by MWM was significantly increased (P<0.05), the protein and gene expressions of Bax and Caspase-3 were significantly reduced (all P<0.001), the protein and gene expressions of Bcl-2 were significantly increased (both P<0.001), and the number of neuron apoptosis was significantly reduced (P<0.001) in the EA group than in the model group. Conclusion: EA can improve the learning and memory damages induced by type 2 diabetic model rats with CI; the action mechanism may be achieved via anti-apoptosis.

This study was aimed to establish a smear protocol for preparing bone marrow cells and investigate its effect on fluorescence in situ hybridization (FISH) signal. Probe DNA (C-myc, MDM2, STK6) was labeled with Spectrum Green, PromoFluor-555 and PromoFluor-415 by nick translation. Five bone marrow samples were tested by two methods separately. Traditional method: after removing the erythrocytes by hypoosmotic solution, the bone marrow cells were fixed in methanol/acetic acid (3:1). Improved method: erythrocytes were removed using density gradient centrifugation and fixed in methanol. The samples were then fixed again in 2 formaldehyde for 5 min. The FISH signal was assessed by comparing the relative signal intensity of each fluorophore with the autofluorescence background. The results indicated that improved method greatly increased the ratio of fluorescence signal intensity in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel (traditional method: 4.3 ± 0.19, 3.52 ± 0.04, 3.07 ± 0.08; improved method: 9.89 ± 0.41, 7.55 ± 0.5, 5.67 ± 0.18, n = 5, P < 0.01) respectively. The signal intensity increased 2.32, 2.14 and 1.85-fold in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel respectively. In addition, the improved method decreased the split signals [traditional method: (15.8 ± 1.74), (20.42 ± 2.88), (23.2 ± 3.02); improved method: (8.6 ± 1.2), (12.28 ± 1.33), (12.6 ± 2.56), n = 5, P < 0.05]. It is concluded that the improved optimal procedure which facilitates FISH intensity on bone marrow cells is developed, showing potential for wide application in the diagnosis of hematologic diseases.
Bone Marrow Cells ; cytology ; Histocytological Preparation Techniques ; Humans ; In Situ Hybridization, Fluorescence ; methods

BACKGROUND: Adipose-derived stem cells (ADSCs) represent one of the promising cell sources for myocardial regeneration due to their easy accessibility and efficacy in the improvement of cardiac function following myocardial infarction. However, previously reported studies on the underlying mechanism of ADSCs-mediated cardioprotective effect mainly focused on the ADSCs cultured at room air. OBJECTIVE: To test the paracrine actions and anti-apoptotic effect of ADSCs under hypoxic conditions. METHODS: After isolation and culture, neonatal rat myocardial cells were injured by hydrogen peroxide and co-cultured with rat ADSCs under normoxia and hypoxia (10% O2) conditions. Ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in the cell pellet and levels of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), and basic fibroblast growth factor (bFGF) were tested by ELISA. Expression of apoptotic proteins Bax and Bcl-2 were determined by western blot. RESULTS AND CONCLUSION: GSH/GSSG, VEGF, IGF-1, and bFGF were decreased in neonatal rat myocardial cells injured by hydrogen peroxide. ADSCs significantly attenuated hydrogen peroxide-induced myocardial apoptosis by increasing the ratio of GSH/GSSG and the secretion of VEGF, IGF-1 and bFGF. ADSCs also down-regulated Bax expression and up-regulated Bcl-2 expression. To conclude, hypoxic conditions can enhance the anti-apoptosis and cardioprotective effects of ADSCs through the paracrine mechanism.

The permeability of cell membrane was enhanced by exogenous SA in the culture of Taxus cuspidata. Nuclei condense and fragments were observed in some cells by using fluorescent microscope, and the degraded chromosomal DNA was observed by using agarose gel electrophoresis. The changes in soluble proteins of the suspension cultures of Taxus cuspidata induced by salicylic acids were analyzed by using two-dimensional polyacrylamide gel electrophoresis. Comparing with the control, seven new protein spots were found and six protein spots were not found in the cultures grown with SA at 48h. The results obtained showed that SA could induce the expression of some special proteins that might be related with the action of SA in the suspension cultures of Taxus cuspidata.
Cell Culture Techniques ; methods ; DNA, Plant ; genetics ; Electrophoresis, Agar Gel ; Electrophoresis, Gel, Two-Dimensional ; methods ; Gene Expression Regulation, Plant ; drug effects ; Plant Proteins ; genetics ; metabolism ; Salicylates ; pharmacology ; Taxus ; genetics ; metabolism