Microfluidics technology may be an effective method to solve some problems in cryopreservation. This review presents the research progress of microfluidics technology in the field of cell membrane transport properties, cryoprotectant addition and washout and the vitrification for cryopreservation of biological materials. Existing problems of microfluidics technology in the application of cryopreservation are summarized and future research directions are indicated as well.
Cell Membrane ; Cryopreservation ; Cryoprotective Agents ; Membrane Transport Proteins ; Microfluidics

Enhancing medical humanistic quality education is the essence of medicine and it is also a necessity of conforming to the transformation of medical model.The pathological teachers should improve their humanistic quality actively,integrate humanistic quality training into pathology teaching and focus on training the social responsibility and healthy psychological qualities in order to enhance medical students' humanistic accomplishment and to make them become high qualified medical talents.

Objective To study the factors influencing outcome of acute arterial embolism in extremity.Method From Aug 1997 to Nov 2001, we have accomplished treatment in 41 cases of acute arterial embolism in extremity with Fogarty catheter.Result Among 41 cases,26 cases were succeeded in leg salvage,8 cases were subject to amputation and 7 cases were dead finally.Conclusions Durationand degree of ischemia in extremity,embolectomy completeness in the case of introoperation,reperfusive injury and myonephropathic-metabolic syndrome,Primary diseases and general conditions are main influencing factors in treatment of acute arterial embolism.

Objective To study the operation methods and efficacy of joint prosthesis with artificial diaphysis for the treatment of tumors at the end of bone. Methods 28 patients with tumors at the end of bone had resection and limb salvage by joint prosthesis with artificial diaphysis. These cases consisted of 11 osteosarcomas, 10 bone giant cell tumors, 2 chondrosarcomas, 2 metastatic carcinomas, 1 malignant fibrous histiocytoma,1 aneurysmal bone cyst and 1 osteochondroma. Among them, there were 7 cases of proximal femoral tumors, 6 cases of distal femoral tumors, 10 cases of proximal tibial tumors, 5 cases of proximal humeral tumors. Results The follow-up periods were 16 months to 15 years. Eighteen patients died. Functional activity was satisfactory in 71% of the patients(2 years). Conclusions Joint prosthesis with artificial diaphysis for the treatment of tumors at the end of bone is feasible, and a better system for reattachment of the soft tissues is needed to avoid pain and a persistent limp.

AIM: To study the signal pathway involved in up-regulation of LPS-induced HO-1 expression by CCK-8. METHODS: Forty-two SD rats were divided into 7 groups (six rats each) randomly as follows: control group, LPS group, LPS+SP600125 (JNK-specific inhibitor) group, CCK-8+LPS group, CCK-8+LPS+SP600125 group, CCK-8 group and CCK-8 +SP600125 group. Lungs from the rats in these 7 groups were excised 6 h after the agents were administered. HO-1 mRNA expression was examined by RT-PCR. The protein expression of HO-1 was detected by Western blotting and immunofluorescence flow cytometry (FCM). RESULTS: There were significant positive expression of HO-1 mRNA in LPS group compared to control group. CCK-8 enhanced LPS-induced HO-1 mRNA expression and CCK-8 alone induced HO-1 mRNA expression as well. The mRNA expressions of HO-1 in LPS group, CCK-8+LPS group and CCK-8 group were 3.01 (P<0.01), 5.88 (P<0.01) and 3.45 (P<0.01) times as many as that in control group, respectively. SP600125 inhibited the mRNA expression of HO-1 induced by CCK-8 and (or) LPS. The change of HO-1 protein expression was in accordance with that of HO-1 mRNA expression by Western blotting and immunofluorescence FCM. CONCLUSION: These results suggest that JNK/c-Jun signal pathway plays an important role in the up-regulation of LPS-induced HO-1 expression by CCK-8.

AIM: To investigate the role of hydrogen sulfide (H_2S) in the cholecystokinin octapeptide (CCK-8) attenuating lipopolysaccharide (LPS)-induced lung injury. METHODS: A rat model of lung injury induced by intravenous injection of LPS was developed. Male Wistar rats were divided into normal control group, LPS group, LPS+CCK-8 group and CCK-8 group. Six hours after LPS injection, partial pressure of oxygen in the arterial blood (PaO_2), H_2S content and cystathionine-γ-lyase (CSE) activity in lung tissue were detected. The mRNA expression of CSE in lung tissue was determined by RT-PCR;the structure of lung tissues was observed under optical microscope. RESULTS: Compared to normal control rats, the LPS-treated rats had significantly decreased PaO_2 level, increased index of quantitative assessment (IQA) score, while H_2S content, CSE activity and the mRNA expression of CSE in lung tissue were significantly increased (all P<0.05). Administration of CCK-8 into LPS-treated rats increased the PaO_2 level and alleviated the degree of lung injury (measured by IQA score). In addition, CCK-8 decreased H_2S content, CSE activity, and the mRNA expression of CSE (all P<0.05). No significant difference of the above-mentioned parameters between CCK-8 group and normal control group was observed. CONCLUSION: CCK-8 reduces LPS-induced lung injury through inhibiting the generation of endogenous H_2S.

Objective To observe the role and mechanism of CO-releasing molecules (CORMs) -2in the injured lung induced by ischmia-reperfusion (IR) of hind limbs of rat.Methods The rat model of lung injury was made by ischemia in hind limbs of rat for two hours and then reperfusion for two hours as well.There were 40 SD rats randomly ( random number) divided into 5 groups ( n =8 ),namely sham ischemia-reperfusion (I/R) group,sham I/R + CORM-2 group,I/R group,I/R + CORM-2 group and I/R + DMSO (Dimethylsulfoxide) group. Rats in sham I/R group underwent laparotomy without infrarenal aorta occlusion.The lung tissue structure,polymorphonuclear neutrophil (PMN) count,wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content, myeloperoxidase (MPO) activity, intercellular adhesion molecule-1 ( ICAM-1 ),nuclear IκBα degradation and NF-κB activity in the lung were measured.Results Compared with the sham I/R group,the number of PMNs in lung,W/D,MDA content,MPOactivity,ICAM-1 and NF-κB activity significantly increased in I/R group,whereas nuclear IκBα decreased (P ＜ 0.01).Compared with the I/R group,the number of PMNs in lung,W/D,MDA content,MPO activity and ICAM-1 significantly decreased in I/R + COMR-2 group ( P ＜ 0.01 ), while nuclear IkBαincreased. Conclusions These data demonstrate that CORM-2 attenuates limb I/R-induced lung injury by inhibiting ICAM-1 protein,NF-κB pathway and the leukocytes sequestration in the lung following limb I/R in rats,suggesting that CORM-2 could be used as one of the most valuable therapeutic agents.

AIM: To study the role of carbon monoxide (CO) in the mechanism of cholecystokinin-octapeptide (CCK-8) for attenuation of acute lung injury (ALI) induced by lipopolysaccharide (LPS). METHODS: Fifty-six adult male rats were randomly divided into seven groups: control group, LPS group, LPS+ZnPP (a specific inhibitor of HO-1) group, LPS+Hemin (Hm, CO donor) group, CCK-8+LPS group, CCK-8+LPS+ZnPP group and CCK-8 group (n=8 for each). Bronchoalveolar lavage was performed 2 h, 6 h and 12 h respectively after treatments. The numbers of polymorphonuclear leukocytes (PMN) in bronchoalveolar lavage fluid (BALF) was detected. The mortality of rats and the structure of lung tissues were observed. MDA and CO contents in lung tissues were also measured. RESULTS: The mortalities of rats were both zero 2 h and 6 h after agent administration. The mortality of rats was higher than control group 12 h after LPS administration. The mortality of rats in LPS+Hm and CCK-8+LPS group were lower than that in LPS group, and its in LPS+ZnPP and CCK-8+LPS+ZnPP group were lower than that in LPS and CCK-8+LPS group, respectively. Lung injury was observed in LPS group. At the same time the number of PMN, MDA and CO content were higher than those in control group. The degree of lung injury, PMN numbers and MDA content were lower, while CO content in LPS+Hm and CCK-8+LPS group were higher than those in LPS group. However, the degree of lung injury, PMN number and MDA content were higher, CO content were lower in LPS+ZnPP and CCK-8+LPS+ZnPP group than those in LPS and CCK-8+LPS group, respectively. CONCLUSION: CCK-8 attenuates the LPS-induced acute lunginjury by means of anti-oxidation and inhibition of PMN aggregation, which are both mediated by CO.

Objective:To establish a method for isolating tumor infiltrating dendritic cells(TIDC)from mice models of Lewis lung cancer by positive selection using anti-CD11c magnetic beads.Methods:A total of 1.0?10~6 Lewis lung cancer cells were subcutaneously injected into a C57BL/6 mice at the lateral abdominal wall to establish the mouse model of lung cancer.TIDC were isolated positively using anti-mouse CD11c magnetic beads;they were labeled by anti-mouse CD11c,and then the purity of the isolated cells was tested by FACScan flow cytometer.The cells were also double labeled by PE-conjugated MHC-ⅡmAb and FITC-conjugated CD83 mAb or FITC-conjugated CD86 mAb to analyze the phenotype of cells by FACScan flow cytometer.Results:The positive selection using anti-CD11 c magnetic beads isolated(1.73?0.31)?10~6 TIDC from each gram of Lewis lung cancer tissue,which accounted for(2.18?0.29)%of the cells in the tumor tissues.The purity of TIDC was 96.49%.Electron microscope showed that the isolated TIDC had the typical character of DC cells.The positive rates of MHC-Ⅱ,CD83 and CD86 molecules in TIDC surface were(51.25?4.21)%, (3.48?0.34)%and(3.07?0.65)%,respectively.Conclusion:The positive selection using anti-CD11c magnetic beads is highly effective,simple,and economic,and is worth popularizing.

Objective To research the relationship between the gene polymorphism of serum amyloid A protein(SAA)1 and coronary heart disease(CHD).Methods By using PCR-RFLP and sequencing,the gene polymorphism of SAA1 of 183 patients with coronary heart disease and 152 healthy controls were analyzed.Result In the both groups 3 alleles(1.1,1.3,1.5)and 6 genotypes(1.1/1.1,1.1/1.5,1.1/1.3,1.3/1.3,1.3/1.5 and 1.5/1.5)were found.The frequency of 1.5 allele in healthy controls group was notably higher than that in CHD group(P