In order to explore the permissible range of temperatures for cryopreserved platelets, each of the eleven bags of platelets with 5% dimethyl sulfoxide was divided at random into eight aliquot paired groups labeled A, B, C, D, E, F, G, and H respectively. All of the groups were stored at -80℃ for one day. The control group A was stored at -80℃ throughout the experiment, and group B, C, D, E, F, G, H were replaced to the refrigerator at -80℃ after they were stored for 4 hours at -60℃, -50℃, -40℃, -30℃, -20℃, -10℃, and 0℃ respectively. All samples were thawed to test the phosphatidylserine expression, CD62p re expression, slide platelet aggregation test(SPAT), and activated plasma clotting test(aPCT). The above items were monitored in group A hourly when group A was thawed and stored at 22℃ for 4 hours. The results showed that the permissible storage temperature of cryopreserved platelet was under -40℃, and storage at temperature from -30℃ to 0℃ was detrimental for cryopreserved platelets. No significant damages were detected when the thawed platelets were stored at 22 with a gentle shaker.

To observe the regulative effect of electroacupuncture(EA) on plasma levels of some brain gut peptides in dogs, contents of gastric secretin(GT), vasoactive intestinal peptide(VIP), somatostatin(SS) and endothelin(ET) in plasma were simultaneously measured by radioactive immunonassay method in the dogs with or without EA stimulation at different acupoints. The results showed that in the Zusanli(st 36) group, after EA stimulation, an increase was observed in plasma contents of GT and VIP( P

Objective To investigate the regulation of electroacupuncture(EA)on gastric mucosal blood flow(GMBF) and its relation to the contents of plasma and gastric mucosal calcitonin gene-related peptide(CGRP) in dogs.Methods Twenty dogs were randomly divided into four groups:blank control group,no meridian point group,Shangjuxu group and Zusanli point group.By using laser doppler flowmeter(LDF)and radioimmunoassay method,GMBF,contents of CGRP were simultaneously measured,with or without EA stimulation at different acupoints in expermental dogs.Results In the Zusanli group,after EA stimulation, an increase was observed in plasma CGRP content(P

Objective 1) To establish and optimize a flow cytometry method for detecting CD62p reexpression by preserved platelets.2) Daily determination of the CD62p expression by platelets stored at 22℃ to assess its value in evaluation of preserved platelet quality.Methods 1) Statistic methods including factorial experiment was applied to optimize the major conditions for sample management, and the feasible negative and positive control for FCM analysis of CD62p expression were check out.2) Thirteen donors' platelets including ten PRP and three apheresis concentrations were stored in a flatbed shaker at 22?2℃ for 12 days. The CD62p expression were detected daily. The correlation between CD62p expression and in vivo residual life span of 22℃ stored platelet were analyzed.Results 1) Satisfactory conditions for analysis CD62p anew expression by FCM were obtained. 2) The CD62p anew expression by stored platelets was correlated to the in vivo residual life span of the platelets with the correlation coefficients between 0.91 and 0.99.Conclusion 1) This FCM method is simple, sensitive, stable, and easy to carry. 2) Platelet CD62p expression can be used to monitor the function of stored platelets and has the potential in the study of new platelet preservation technique.

Objective To observe the effect of early non-invasive positive pressure ventilation(NIPPV)on the treatment of serious hypoxemia induced by acute left heart failure. Methods Forty patients with acute left heart failure( Grade Ⅳ heart function)were randomly divided into two groups. Patients in both groups accepted supportive treatment included cardiotonics, diuretics, vasodilators, in additional to these high concentrations of oxygen were given in conventional group, and non-invasive positive pressure ventilation were given in NIPPV group by biphasic positive airway pressure(BiPAP). Systolic blood pressure, heart rate, respiratory rate, blood-gas analysis( pH, PaO2, PaCO2, SaO2 )and clinical signs were observed at 2 hours after treatments. Results Compared to control,RR( [ 19.55 ± 1.88] vs [21.85 ±3.51 ] ) BPM] ,HR ( [96.40 ±2.80] vs[ 103.20 ±6.78 ] BPM), SBP ( [ 116.30 ± 8.95 ] mm Hg vs [ 122.50 ± 6.13 ] mm Hg), pH (7.404 ± 0.027 vs 7.358 ±0.05) ,SaO2 ( [93.57 ± 1.18]% vs [91.97 ± 1.85]% ) ,PaO2 ( [75.58 ±4.61 ]mm Hg vs [68.38 ±7.95]mm Hg), PaCO2 ( [ 38.69 ± 3.06 ] mm Hg vs [ 43.61 ± 2.65 ] mmHg) were significantly different in NIPPV group( t = 2.582,4.146,2.558,3.534,3.256,3.505,5.428, Ps ＜ 0.05 ). We found no significant differences in the comparisons before treatments. Hypoxia improved in NIPPV group,and the total effective rate was 95% in NIPPV group and 70% in control group,which showed significant difference( x2 =4.329 ,P ＜0.05 ) Conclusion BiPAP non-invasive positive pressure ventilation combined with routine treatment in treating heart failure, could promptly correct hypoxia, improve heart function and shortening disease course.

To explore the mechanism of electroacupunture (EA) of Zusanli (ST36)in protecting gastric mucosa of dogs.Twenty mongrel dogs were randomly allocated to four groups: blank control group (Group A), non acupoint group (Group B), Shangjuxu (ST37) group (Group C) and Zusanli (ST36) group (Group D). Dynamic gastric mucosal blood flow (GMBF) was monitored by laser Doppler flowmeter and calcitonin gene related peptide (CGRP) contents in plasma and gastric mucosa were measured simultaneously by radioimmunoassay method. After sixty minutes of EA, GMBF and CGRP contents in plasma and gastric mucosa were increased in Group D (P

Objective To investigate the clinical effects of the posterior atlantoaxial pedicle screw fixation combined with unilateral axial spinous process and lamina screws and allogeneic bone graft for atlantoaxial instability.Methods From March 2010 to April 2014,data of 10 patients with atlantoaxial instability who had undergone atlanto-axial vertebral pedicle fixation and interbody fusion combined with posterior atlanto-axial spines lamina nails were retrospectively analyzed.There were 6 males and 4 females with an average age of 39 (range,16-62) years old.The inclusion criteria was atlantoaxial instability together with unilateral vertebral artery segment high cross deformity (7 patients) or atlantoaxial joint break bad involving unilateral pedicle (7 patients).3 patients have spinal cord disease symptoms and physical signs,both suffered limited neck mobility and pain.The VAS scores were 1-8,with an average of 3.70±2.11.Preoperative X-ray,CT three-dimensional reconstruction and MRI were collected.X-ray and CT examination were performed 7 d and 1,3,6,12 months after operation to evaluate the internal fixation and bone grafting fusion.Results There's no occurrence of cervical spinal cord and vertebral artery injury.Screws were successfully implanted.1 case's incision occurred infiltration liquid 3 days after operation.For allograft rejection consideration,we changed the plan,and the infiltrating stopped 5 days later.Other incisions were all primary healing.VAS scores were 1.01±0.89 after operation,which was significantly reduced.X-ray showed good recovery of cervical sequence after the operation,and CT revealed 1 patient with atlas pedicle screw in the medial cortex damage.Spinal canal was without infringing,and the other screw positions were normal.6 months after operation,X-ray or CT examination both showed bony fusion.Conclusion Atlantoaxial pedicle screw fixation combined with unilateral axial spinous process and lamina screws and allograft bone graft fusion for the treatment of atlantoaxial instability can observe good clinical effects.

Objective To evaluate the internal irradiation biological effects of 125I-UdR chitosan nanoparticles in hepatoma cells.Methods The accumulation and distribution of 125I-UdR-CS-DLN in hepatoma cells HepG2 and human liver tissue cells HL-7702 were observed with a confocal microscopy.The internal irradiation biological effects were evaluated by MTT assay,flow cytometry and single cell gel electrophoresis.The apoptosis of in situ rabbit liver tumor treated with 125I-UdR-CS-DLN was assayed by TUNEL staining technique.Results After 30 min of nano-particle treatment,its accumulation in the cytoplasm of HepG2 cells was significantly greater than that in HL-7702 cells.When the concentrations of 125I-UdR-CS-DLN was higher than 37 kBq/ml,the cell viability of HepG2 was significantly lower than that of lL-7702 at 24 and 48 h post-treatment(t =-4.46-6.31,P＜0.05),and the HepG2 cells were arrested at G1 phase and significantly impaired at G2/M phase.In addition,the degrees of DNA doublestrand break of both cell lines irradiated by 125I-UdR-CS-DLN were significantly higher than those treated with 125I-UdR,and the DNA repair capacity of HepG2 cells was significantly lower than that of HL-7702 cells(OTM:t =2.94,P ＜0.05;TDNA％:t =10.64,P ＜0.01).TUNEL staining showed that cell apoptosis could be induced in the rabbit liver carcinoma by 125I-UdR-CS-DLN but not by 125I-UdR.Conclusions The amount of 125I-UdR-CS-DLN absorbed by hepatoma cells is significantly higher than that of 125I-UdR,which suggests that 125 I-UdR-CS-DLN induces more stronger internal radiation biological effects of apoptosis and DNA damage on hepatoma cells.

The temperature of platelet intracellular ice crystal formation (IIF) is one of the most important physical parameters to instruct platelet cryopreservation. In this study, the range of temperatures for platelet IIF was measured by means of biological and physical methods. All platelet samples were graded cooling, and two samples of per 5 degrees C decrease were thawed by 2 different ways: 37 degrees C directly (T 37 degrees C) and 37 degrees C after keeping in liquid nitrogen (LN) for 2 hours. The phosphatidylserine (PS) positive rate, plasma lactate dehydrogenase (LDH) concentration and platelet aggregate rate were measured in all samples. The heat release graphs of platelets cryopreserved with or without 5% DMSO were also measured by differential scanning calorimeter (DSC). The results showed that the PS positive rates and aggregate rates in platelets and plasma LDH concentrations gradually increased in T 37 degrees C group and decreased in LN group until the arrival of -35 degrees C, and then there were no further changes of the 3 parameters. A small second heat release peak was detected at about -35 degrees C in the platelet samples cryopreserved without DMSO. It is concluded that the temperature of intracellular ice crystal formation in platelet is from -30 to -40 degrees C (-35 degrees ).
Blood Platelets ; physiology ; Blood Preservation ; Cryopreservation ; Crystallization ; Humans ; Temperature

Human platelets have distinct characters when preserved by different methods. A efficient flow cytometric assay for different preserved platelets expression of CD62p and phosphatidylserine(PS) is in dire need. Efficient flow cytometric assay for CD62p and PS expressed by preserved platelets was established and the major conditions were optimized. The platelets need not to be washed to wipe off plasma and can be labelled diredtly during the sample preparation. It is efficient for flow cytometric analysis when fresh platelet riched plasma (FPRP) was set as negative control, thrombin actived FPRP, and liquid nitrogen treated FPRP were set as positive control respectively. Gly-Pro-Arg-Pro acetate salt (GPRP) was applied to prevent platelets aggregation and fibrin formation, stabilize platelets and minimize the artificial platelets activation. This is also the key to conquer difficulty of flow cytometric quantitive analysis when platelet, Ca(2+) and plasma coexist. This flow cytometric method is specially suitable for the multi-parameter assay including PS expression for cryopreserved platelets. Minimal sample manipulation, no fixation, and GPRP application resulted in minor artifacts and good sample stability. Results suggested, this flow cytometric assay for preserved platelets is simple and efficient. In addition, the author prepared four different methods treated platelets that can be easily distinguished through this flow cytometric assay. It not only makes sure the practicability of this flow cytometric assay, but also suggests the value of the treated platelets applied in preserved platelets flow cytometric ass
Blood Platelets ; metabolism ; Flow Cytometry ; methods ; Humans ; P-Selectin ; biosynthesis ; Phosphatidylserines ; analysis ; Reproducibility of Results ; Tissue Preservation