A mini review of Liparis species: an ornamental genus with considerable medicinal value.

The genus Liparis, a group of perennial ornamental herbs in the family Orchidaceae, is widely distributed in tropical and subtropical regions. Many species of the genus Liparis have been commonly used as traditional herbal medicines for the treatment of menorrhagia, haemoptysis, traumatic bleeding, snake bites, and pneumonia. This review describes the ornamental value of plants of the genus Liparis and summarises the chemical constituents and pharmacological activities reported during the last decade. The main chemical constituents of this genus are phenolic acids, alkaloids, flavonoids, etc. Most phenolic acids and alkaloids have a nervogenic acid skeleton, and most alkaloids also have a pyrrolizidine skeleton. Extracts from the genus Liparis plants showed significant haemostatic, antitumor, anti-inflammatory, hypolipidemic, antioxidant, and antibacterial activities. This paper proposed ideas and research directions for the future study of plants in the genus Liparis, providing valuable information for the development of new drugs and promoting their utilisation.

[Manifestations and distribution rules of jingjin lesions in neck-type cervical spondylosis].

OBJECTIVE: To explore the manifestations of jingjin (sinews/fascia) lesions and summarize their distribution rules in the patients with neck-type cervical spondylosis so as to provide the evidences for the development of clinical diagnosis and treatment scheme of acupuncture for cervical spondylosis. METHODS: A total of 120 patients with neck-type cervical spondylosis were collected. The meridian diagnostic method was used to examine the upper back of each patient, the manifestation category of jingjin lesions, locations and the affected muscle regions of twelve meridians were recorded. RESULTS: (1) The punctate lesions of jingjin were detected in 15 regions, and the highest frequency of lesion occurred in the region from the inner upper corner of the scapula to Quyuan (SI 13) (113 cases, 94.2% of lesion frequency). The lesion frequency of 10 regions was ≥50.0%. The punctate lesions were mainly distributed in the muscle regions of hand-shaoyang (349 cases) and foot-taiyang (333 cases). (2) The linear lesions of jingjin were detected in 10 regions, and the highest frequency of lesion occurred in the region from the inner upper corner of the scapula to Quyuan (SI 13) (77 cases, 64.2% of lesion frequency). The lesion frequency of 2 regions was ≥50.0%. The linear lesions occurred mainly in the muscle region of foot-taiyang (251 cases). (3) Eight regions were examined to be the planar lesions of jingjin, and the highest frequency of lesion was found in the site of Jianjing (GB 21) (84 cases, 70.0% of lesion frequency). The lesion frequency of 3 regions was ≥50.0%. The muscle region of foot-taiyang (260 cases) was predominated in the planar lesions. (4) The distribution of all of the punctate, linear and planar lesions of jingjin was analyzed statistically. It was found that 25 regions were involved and those with the high lesion frequency were distributed in the area from the inner upper corner of the scapula to Quyuan (SI 13), the sites of Jianjing (GB 21) and Dazhui (GV 14), transverse processes of C3 to C5 and the area from the lateral border of the scapula to the teres minor, separately. The muscle regions of foot-taiyang, hand-shaoyang and hand-yangming were involved in various kinds of jingjin lesions. CONCLUSION: Jingjin lesions in patients with neck-type cervical spondylosis can be divided into three categories, namely, punctate, linear and planar lesions; of which, the punctate lesions are dominated. A majority of jingjin lesions is related to the muscle region of foot-taiyang, and the lesion frequency is higher compared with the lesions to the muscle regions of hand-shaoyang and hand-yangming. Jingjin lesions are commonly distributed in the area from the inner upper corner of the scapula to Quyuan (SI 13).

Effect of large volume red blood cell apheresis on cardiovascular functions in healthy donors.

BACKGROUND: Requirements of blood transfusions rise rapidly in China. Improving the efficiency of blood donation could help maintaining sufficient blood supplement. We conducted a pilot research to investigate the reliability and safety of collecting more units of red blood cell by apheresis. METHODS: Thirty-two healthy male volunteers were randomized into two groups: red blood cell apheresis (RA) (n = 16) and whole blood (WB) donation (n = 16). RA group donated individualized RBC volumes by apheresis according to the volunteers' basal total blood volumes and haematocrit levels, WB group donated 400 mL whole blood. All volunteers were scheduled seven visit times in 8 weeks' study period. The cardiovascular functions were assessed by laboratory examinations, echocardiography and cardiopulmonary functional tests. All results were compared between groups at the same visit time and compared between visit 1(before donation) and other visit times within the same group. RESULTS: The average donated RBC volume in RA group and in WB group was 627.25 ± 109.74 mL and 175.28 ± 8.85 mL, respectively(p < 0.05); the RBC, haemoglobin and haematocrit levels changed significantly between times and between groups (p < 0.05). Cardiac biomarker levels such as NT-proBNP, hs-TnT and CK-MB did not change significantly between times or between groups (p > 0.05). The echocardiographic and cardiopulmonary results did not change significantly between times or between groups during the whole study period(p > 0.05). CONCLUSIONS: We provided an efficient and secure method for RBC apheresis. By harvesting more RBC volumes at one single-time, the cardiovascular functions did not change significantly compared with traditional whole blood donation.

[Effect of meridian sinew releasing technique on moxibustion sensation of heat-sensitive moxibustion in patients with knee osteoarthritis].

OBJECTIVE: To observe the effect of meridian sinew releasing technique on moxibustion sensation of heat-sensitive moxibustion in patients with knee osteoarthritis (KOA). METHODS: A total of 60 patients with KOA were randomly divided into an observation group and a control group, 30 cases each group. In the observation group, on the basis of the meridian sinew releasing technique, moxibustion sensation exploration method was applied at Dubi (ST 35) area on the affected side. In the control group, moxibustion sensation exploration method was applied at Dubi (ST 35) area on the affected side. The meridian sinew releasing technique was performed for 20 min each time, the moxibustion sensation exploration method was performed for 60 min each time, once a day for 3 days. The excitation rate, latency, duration time and intensity value of moxibustion sensation of heat-sensitive moxibustion were recorded on the 1st, 2nd and 3rd days of exploration in the two groups. RESULTS: The excitation rate on the 3rd day of exploration and total excitation rate in the observation group were higher than the control group (P<0.05). On the 1st, 2nd and 3rd days of exploration, the latency of moxibustion sensation of heat-sensitive moxibustion in the observation group was shorter than the control group (P<0.05), the duration time was longer than the control group (P<0.05), and the intensity value was higher than the control group (P<0.05). CONCLUSION: Meridian sinew releasing technique could improve the excitation rate of moxibustion sensation of heat-sensitive moxibustion in patients with KOA, shorten the latency, prolong the duration time, and improve the intensity value.

Phytochemical constituents of Camellia osmantha fruit cores with antithrombotic activity.

Camellia osmantha is a new species of the genus Camellia and is an economically important ornamental plant. Its activity and ingredients are less studied than other Camellia plants. This study investigated the antithrombotic effect and chemical components of C. osmantha fruit cores using platelet aggregation assays and coagulation function tests. The cores of C. osmantha fruits were extracted with ethanol to obtain a crude extract. The extract was dissolved in water and further eluted with different concentrations of methanol on an MCI resin column to obtain three fractions. These samples were used for antithrombotic activity tests and phytochemical analysis. The results showed that the extract and its fractions of C. osmantha have strong antithrombotic activity, significantly reducing the platelet aggregation rate and prolonging the thrombin time (TT). The total saponins, flavonoids, and polyphenols in the active fractions may be responsible for the antithrombotic activity. The chemical constituents were analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). Twenty-three compounds were identified rapidly and accurately. Among them, ellagic acid, naringenin, and quercetin 3-O-glucuronide may be important antithrombotic constituents. Furthermore, interactions between these compounds and the P2Y1 receptor were investigated via molecular modeling, because the P2Y1 receptor is a key drug target of antiplatelet aggregative activity. The molecular docking results suggested that these compounds could combine tightly with the P2Y1R protein. Our results showed that C. osmantha fruit cores are rich in polyphenols, flavonoids, and saponins, which can be developed into a promising antithrombotic functional beverage for the prevention and treatment of cardiovascular and cerebrovascular diseases.

Acetylated Rhamnose Triterpenoid Saponins from Glechoma longituba Analyzed by LC-Q-TOFMS.

The aim of the present work is to isolate a series of triterpene derivatives with rhamnosyl linking acetyl groups from Glechoma longituba according to the structural characteristics of previously described triterpene saponins. The extract ion chromatography spectrum of the crude extract of G. longituba was detected and analyzed by HPLC-HR-ESI-MS to determine possible components, and these metabolites were traced and separated by combining high-resolution mass spectrometry and predicted liquid chromatography retention time. Three 11α, 12α-epoxypentacyclic oleanolic acid triterpene saponins (glechomanosides H-J) and one ursane triterpene aldehyde saponin with a C-28 aldehyde group were isolated from G. longituba. The structure of these compounds was confirmed by NMR and compared with those of previously characterized compounds. The strategy described in this report enables a rapid, reliable, and complete analysis of glycoside compounds containing different numbers of acetyl groups at different positions on the sugar.

Antitumor activity on human bladder cancer T-24 cells and composition analysis of the core of Camellia osmantha fruit.

In this study, antitumor activity and composition analysis of the core of Camellia osmantha fruit were investigated. The cores were extracted by 80% ethanol to obtain the crude extract (TF), which was successively distributed by different concentrations of ethanol with MCI resin to obtain three different parts (Frs 1 to 3). Cytotoxicity activities showed that TF, Fr-2, and Fr-3 exhibited good inhibition against T-24 with IC50 values of 6.7 ± 1.3, 6.9 ± 0.9, and 6.7 ± 1.7 µg/mL, respectively. Fr-3 has a strong inhibitory activity on T-24 cells, mainly due to effectively increasing the release of intracellular calcium ions and reactive oxygen species. In addition, Fr-3 can inhibit T-24 cells migration and invasion. Further composition analysis on Fr-3 was detected by LC-Q-TOFMS implying the main components to be ellagic acid and its derivatives.

Five 11α, 12α-epoxy pentacyclic triterpenoid saponins with antithrombus activities from Glechoma longituba.

The present study reports the phytochemical investigation of n-butanol-soluble extracts of Glechoma longituba. Five new oleanane-type triterpenoid saponins with an 11α, 12α-epoxy unit, named glechomanosides A - E, were isolated from the n-butanol soluble fraction of G. longituba. Their chemical structures were established using HRESIMS, IR, 1D NMR, and 2D NMR techniques. The compounds were all evaluated for their antithrombus activities by monitoring thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (APTT), and antiplatelet aggregation assays. These results suggest that G. longituba might be a candidate plant source of an interesting antithrombotic activity.

NF-κB inhibitory and cytotoxic activities of hexacyclic triterpene acid constituents from Glechoma longituba.

BACKGROUND: Non-Small-Cell Lung Cancer (NSCLC) is the most-frequent cause of cancer death, and novel chemotherapeutic drugs for treating NSCLC are urgently needed. 2α, 3α, 23-trihydroxy-13α, 27-cyclours-11-en-28-oic acid (euscaphic acid G) is a new hexacyclic triterpene acid isolated by our group from Glechoma longituba (Nakai) Kupr. However, the underlying mechanisms responsible for the anticancer effects of hexacyclic triterpene acid have not been elucidated. PURPOSE: In the present work, we evaluated growth inhibitory effect of the new isolated hexacyclic triterpene acid and explored the underlying molecular mechanisms. METHODS/STUDY DESIGNS: Herbs were extracted and constituents were purified by chromatographic separation, including silica gel, ODS, MCI, Sephadex LH-20 and preparative HPLC. The compound structures were elucidated by the use of UV, NMR and MS spectral data. The anticancer activity of euscaphic acid G was evaluated by MTT assay. Cell cycle, apoptosis, reactive oxygen species and mitochondrial membrane potential were determined by flow cytometry. To display the possible mechanism of euscaphic acid G on NCI-H460 cells, RT-PCR, immunofluorescence and Western blot analysis were carried out. RESULTS: A new hexacyclic triterpene acid, euscaphic acid G, together with fifteen known triterpenoids, was isolated from the aerial parts of G. longituba. Our results showed that euscaphic acid G exerted strong anti-proliferative activity against NCI-H460 cells in a concentration- and time-dependent manner. Flow cytometry demonstrated euscaphic acid G arrested the cell cycle at G1 phase, induced cellular apoptosis, accompanied by ROS generation and mitochondrial membrane potential reduction. Mechanistic studies revealed that euscaphic acid G treatment inhibited IKKα/ß phosphorylation and IκBα phosphorylation, which subsequently caused the blockage of NF-κB p65 phosphorylation and nuclear translocation. CONCLUSION: In conclusion, these results suggested that euscaphic acid G from G. longituba showed potential anticancer effects against lung cancer cells via inducing cell cycle arrest and apoptosis, at least partly, through NF-κB signaling pathways.

Astaxanthin Combine with Human Serum Albumin to Abrogate Cell Proliferation, Migration, and Drug-resistant in Human Ovarian Carcinoma SKOV3 Cells.

BACKGROUND: Astaxanthin (AST) shows a large range of beneficial effects together with anti-cancer and antioxidation properties. Human Serum Albumin (HSA) is the most abundant protein in blood plasma which plays the role of a depot and transport protein for many exogenous compounds. However, whether HSA could enhance AST-induced cytotoxic effects in human ovarian cancer cells has not been examined to date. OBJECTIVE: This study aims to explore the anticancer effect and the molecular mechanism of AST combine with HSA induced cytotoxicity in ovarian cancer SKOV3 cells. METHODS: The ovarian cancer SKOV3 cells were treated by AST combined with HSA to study the effects of cell proliferation, cell morphology, cell cycle arrest, related protein expression, nuclear transfer, cell migration, and drug-resistant. RESULTS: Our data confirmed that AST+HSA treatment enhanced the anticancer effects of AST, arrested G1 phase cell cycle and induced apoptosis in SKOV3 cells. AST+HSA induced apoptosis via mitochondrial apoptotic pathways was related to the increased ratio of Bcl-2/Bax and activation of caspase-3. Besides, exposure of cells to AST+HSA triggered the inactivation of NF-κB and activation p53 and MAPKs signaling pathways. Furthermore, AST+HSA significantly overcome the drug-resistant and inhibited the migration of SKOV3 cells. CONCLUSION: AST combined treatment with HSA considerably inhibited NF-κB expression and translocation to nucleus, thereby improving the AST-induced cytotoxic effect on SKOV3 cells. These findings may provide rationale to combine AST with HSA for the treatment of ovarian cancer.

New inhibitors of matrix metalloproteinases 9 (MMP-9): Lignans from Selaginella moellendorffii.

Matrix metalloproteinase 9 (MMP-9) is one of the structurally related zinc-dependent endopeptidases families and provides a new target for cancer therapy owing to its pivotal role in metastatic tumors. In this paper, fourteen lignans, including three novel lignans, named selamoellenin B-D (1-3), and eleven known lignan derivatives (4-14) were isolated from the plant of Selaginella moellendorffii. Among them, compound 3 is optically active, which was enantiomerically seperated to afford a pair of enantiomers, (-)-3 and (+)-3. Their structures were elucidated by extensive spectroscopic analyses. Their cytotoxic activities were evaluated against four human cancer cell lines. Among them, five compounds (4, 5, 6, 11 and 13) exhibited great potent cytotoxicity and their structure-activity relationships were also discussed. All compounds except for 3 lignan analogues with low cytotoxicity were selected for further in vitro enzyme inhibition, surface plasmon resonance (SPR), and molecular docking assays based on the MMPs target. The results shown that, compound 11 have the best inhibitory effect and can be considered as a potential drug candidate targeting at MMP-9 for cancer therapy.

6-Chloro-2-phenyl-3-(2-phenyl-ethyn-yl)quinoxaline.

In the title compound, C(22)H(13)ClN(2), the quinoxaline ring system is close to planar [maximum deviation = 0.061 (2) Å]. The phenyl ring at the 2-position and the phenyl ring of the phenyl-ethynyl substituent make dihedral angles of 49.32 (7) and 11.99 (7) °, respectively, with the quinoxaline mean plane. The two phenyl rings are inclined to one another by 61.27 (9)°. In the crystal, mol-ecules are linked by C-H⋯π and π-π inter-actions [centroid-centroid distances = 3.6210 (12) and 3.8091 (12) Å].

[Optimization of lyophilization procedures for freeze-drying of human red blood cells].

OBJECTIVE: To investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells (RBCs). METHODS: Human RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 degrees celsius;. The morphological changes of the RBCs were observed to assess the degree of vitrification, and the specimens were placed in the freeze-dryer with the temperature set up at 40, -50, -60, -70 and -80 degrees celsius;. The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared, and the water residue in the specimens was determined. RESULTS: The protectant did not show ice crystal in the course of freezing and thawing. No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of -70 degrees celsius; and -80 degrees celsius; (P > 0.05). The E procedure resulted in the maximum recovery of the RBCs (83.14% ± 9.55%) and Hb (85.33% ± 11.42%), showing significant differences from the other groups(P < 0.01 or 0.05). The recovery of the RBCs showed a positive correlation to the water residue in the samples. CONCLUSION: Fast cooling in liquid nitrogen and shelf precooling at -70 degrees celsius; with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.

[Preliminary study on rehydrated conditions for lyophilized human red blood cells].

The objective of this study was to investigate the effect of different rehydration conditions on recovery of the lyophilized red blood cells (RBC) so as to optimize the RBC rehydration. The different conditions, including different rehydration solution, the rehydration temperature, volume change rate of the lyophilized RBC rehydrated by the vapor firstly, were studied, the recovery rate and change of physiological and biochemical properties of the rehydrated RBC were detected. The results indicated that the solution of 10% (w/v) PVP40 in PBS showed the best effect, and the RBC recovery rate increased with increasing of rehydration temperature, and the optimal temperature of rehydration was at 37 degrees C. Pre-rehydration in condition of vapor could raise the RBC recovery rate, and promote the MCV and RDW to close to index of the fresh RBC, the deformability of the rehydrated RBC was no serious as compared with RBC preserved in conventional condition, but the activity level of ATP, G-6-PD, SOD, 2, 3-DPG of the rehydrated RBC less decreased. It is concluded that the optimal rehydration conditions for lyophilized RBC are pre-rehydration in the 37 degrees C with vapor firstly, PBS + 10% (w/v) PVP40 rehydration solution and rehydration temperature at 37 degrees C, but the protection of RBC membrane needs to be furtherly studied.

[Optimization of composition and concentration for lyophlizing protectant of human red blood cells].

This study was purposed to investigate the effect of different compositions and concentrations of lyophilizing protectants on recovery of RBCs and hemoglobin (Hb) after rehydration of lyophilized RBCs. The RBC lyophilizing protectants composed of a series concentrations of PVP, trehalose and different osmotic protectants were applied for protecting lyophilizing process of RBCs, the recovery of RBCs and Hb after rehydration of lyophilized RBCs was detected. The results showed that there were significant differences in loss ratio of RBCs between protectants composed of different compositions and concentrations (p<0.05 or p<0.01). The loss ratio of RBCs in protectant containing 30% PVP40, 150 mmol/L trehalose and 2% BSA was minimum (0.02%), the loss ratio of RBCs in protectant containing 6% PVP 360, 100 mmol/L trehalose and 2% BSA was maximum (0.27%). The difference of effect between 150 and 50 mmol/L trehalose was statistically significant (p<0.01). The recovery rates of RBCs and Hb in protectants contained PVP40 of different concentrations were different after rehydration of lyophilized RBCs. The protectant containing 15% PVP40, 150 mmol/L trehalose and 2% BSA showed optimal protective efficacy for lyophilized RBCs, the recovery rates of RBCs and Hb were 61.29+/-4.11% and 62.49+/-5.91% respectively, which were statistically different from other protectants (p<0.01). The protectants containing glycerol displayed best efficiency in lyophilization too, the recovery rates of RBCs and Hb were 65.97+/-4.52% and 67.24+/-5.94%, respectively. It is concluded that the protectants composed of 0.8 mol/L glycerol, 15% PVP40, 150 mmol/L trehalose and 2% BSA (pH 7.3 ) may be used as the protectant lyophilizing human RBCs in future study.

[Cryopreservation strengthens procoagulative activities of platelets].

The aim of this study was to explore the potential relationship between the enhancement of instant hemostatic function in vivo of cryopreserved platelets and its procoagulative related molecule activities. The ability of platelet binding factor V density of GPIb-IX-V (CD42a) at platelet member surface were detected by flow cytometry, the clotting time induced by activated platelets were evaluated by coagulometer and platelet count, MPV and PDW were measured by hemocytometer before and after fresh platelets were cryopreserved. The results showed that the clotting time induced by activated cryopreserved platelets decreased by 43.9%, even quicker than that induced by fresh platelets; the fluorescence intensity of cryopreserved platelet binding factor V increased by 117%, more than that of fresh platelets binding factor V; the GPIb-IX-V (CD42a) density at cryopreserved platelet membrane surface increased by 32%, higher than that at fresh platelet surface. It is concluded that the enhancement of instant hemostatic function in vivo of cryopreserved platelet may be related to higher expression of procoagulative molecules or to their enhanced activity and rapid hemostatic effect.

[Protective effects of DMSO on function of lyophilized human platelets].

This study was aimed to investigate the effects of DMSO on platelets during pre-treatment for lyophilization, including centrifugation, washing and loading trehalose. After pre-treatment for lyophilization, the expression of platelet membrane surface glycoprotein (GP) including CD62p and PAC-1 was analyzed by FCM before and after induction with thrombin, the mean platelet volume (MPV) and platelet maximal aggregation with several platelet inducers were investigated. The results showed that the expression rates of CD62p and PAC-1, as the platelet activation signs, increased and were 30.37% and 15.01% respectively in group without DMSO after pre-treatment. And their differences in comparison with control were statistically significant, but that of CD62p was 10.72% and PAC was 10.11% in group with DMSO, in comparison with group without DMSO respectively, their differences were statistically significant after diluting with DMSO, CD62p was re-expressed to 54.39% in group with DMSO and more than that in group without DMSO and lower than control statistically significant. PAC-1 was re-expressed to 49.28% in group with DMSO and more than that in group without DMSO (p<0.01) and reached to control. Platelet maximal aggregations induced by thrombin, restocetin and propyl gallate were 92.76%, 91.24% and 89.66 respectively in group with DMSO. These were closed to that in control group and in group without DMSO. But the aggregation induced by ADP was 34.33%, it was less than control (p<0.01) and more than that in group without DMSO (p<0.01). It is concluded that DMSO can inhibit the expression of CD62p and PAC-1 on platelet in vitro. But when diluted with plasma, platelets can express CD62p and PAC-1 induced by thrombin and be led to aggregate by several inducers, so the inhibitory effects of DMSO on platelet activation are reversible. DMSO play roles in inhibitor damage from platelet activation and cryoprotectant. This property of DMSO is very important in research of platelets lyophilization.

[Inhibition of L-arginine and cilostazol on activation of platelets in vitro].

The purpose of study was to investigate the effects of L-arginine and cilostazol on platelet-activation and aggregation reserve in vitro, so as to provide proof for selecting reversible activation-inhibitors for platelets lyophilization. Activation and function of platelets were investigated by using flow cytometry with the CD62p and PAC-1 expression and re-expression after being activated by thrombin, and by means of platelet aggregation reaction to thrombin, ADP and propyl gallate, as well as coagulation activity of platelets. The results showed that expression of CD62p and PAC-1 increased after being pretreated. Both L-arginie and cilostazol could inhibit CD62p and PAC-1 expression and related with their concentrations. Cilostazol had an intensive inhibition effect on expressions of CD62p and PAC-1 induced by thrombin, and the inhibition increased when concentration augmented. L-arginine had the same effects on PAC-1, but had no effects on CD62p. L-arginine and cilostazol inhibited aggregation induced by thrombin, ADP and propyl gallate, and the inhibitions were related directly with dosage. When L-arginine concentration was higher or equal to 15 mmol/L, or cilostazol concentration was in range of 1 - 4 mmol/L, the aggregation time were prolonged so much or even no aggregation. It is concluded that when L-Arginine concentration is 5 mmol/L and 10 mmol/L, platelet activation can be inhibited, but aggregation ability and characters keep intact. Concentration at 5 mmol/L may be the best. 1 mmol/L of cilostazol can inhibit activation in vitro and retain part of platelet ability of aggregation and reexpression.

[Experimental study on lyophilization of platelets].

The aim of this study was to search a procedure of platelet lyophilization and find a way of long-term storage of human platelets at normal temperature with smaller size and lighter weight, to be convenient to transport at long distance thus to meet the demands in accidents and war time. Human platelets were pretreated by freezing, the first and the second desiccation, and were added with reversible activation-inhibitors of platelets, DMSO and trehalose, then were rehydrated. At the same time, the recovery rate of platelets, platelet maximal aggregation induced by thrombin, coagulation of platelets, CD62p expression and PAC-1 expression were assayed. The results indicated that the recovery rate of the platelets was 56.29%. The platelet maximal aggregation induced by thrombin had no significant difference between lyophilized platelets and the fresh platelet-rich plasma (FPRP), but the aggregation of platelets induced by ADP or propyl gallate was decreased by 49.34% and 26.25%. Coagulation of the lyophilized platelets was not significantly different from FPRP. CD62p expression of the lyophilized platelets (42.36%) was higher than that in FPRP while PAC-1 expression was 2.12%. CD62p re-expression rate induced by thrombin was 50.88% and PAC-1 re-expression was 54.55%. It is concluded that the ability of recovered lyophilized platelets added with reversible activation-inhibitors, DMSO and trehalose to aggregate and coagulate has showed no significant difference as compared with FPRP. The reversible activation-inhibitors can decrease CD62p expression of lyophilized platelets, and may enhance their survival ability and prolongate survival time. Therefore the efficiency of lyophilizing platelets can be improved based on this freeze-drying procedure.

[Optimization of trehalose loading in red blood cells before freeze-drying].

The key points for better protection of trehalose in freeze-drying red blood cells (RBCs) are to resolve non-osmosis of trehalose to red blood cells and to make cytoplasmic trehalose to reach effective concentration. This study was aimed to investigate the regularity of loading RBCs with trehalose, screen out optimal loading condition and evaluate the effect of trehalose on physico-chemical parameters of RBCs during the period of loading. The cytoplasmic trehalose concentration in red blood cells, free hemoglobin and ATP level were determined at different incubation temperatures (4, 22 and 37 degrees C), different trehaolse concentrations (0, 200, 400, 600, 800 and 1000 mmol/L) and different incubation times (2, 4, 6, 8 and 10 hours), the cytoplasmic trehalose, free hemoglobin (FHb), hemoglobin (Hb) and mean corpuscular volume (MCV) in fresh RBCs and RBCs stored for 72 hours at 4 degrees C were compared, when loading condition was ensured. The results showed that with increase of incubation temperature, time and extracellular trehalose concentration, the loading of trehalose in RBCs also increased. Under the optimal loading condition, cytoplasmic trehalose concentration and free hemoglobin level of fresh RBCs and RBCs stored for 72 hours at 4 degrees C were 65.505 +/- 6.314 mmol/L, 66.2 +/- 5.002 mmol/L and 6.567 +/- 2.568 g/L, 16.168 +/- 3.922 g/L respectively. It is concluded that the most optimal condition of loading trehalose is that fresh RBCs incubate in 800 mmol/L trehalose solution for 8 hours at 37 degrees C. This condition can result in a efficient cytoplasmic trehalose concentration. The study provides an important basis for long-term preservation of RBCs.