This article introduces cryosurgery devices, cryopreservation devices, some cryogenic freezing methods in medicine and the recent progress about cryotherapy technology.
Cryopreservation ; instrumentation ; Cryotherapy ; instrumentation ; methods

This study was performed to evaluate the efficiency of simplified EM grid vitrification, skipping the step of removing the cryoprotectant (5.5M EG + 1.0M sucrose) droplet on the grid after loading oocytes, compared to conventional cryopreservation protocols for mouse mature oocytes. Firstly, the recovery, survival, fertilization and hatching rates of simplified EM grid vitrification were compared with those of the slow freezing method using 1.5M DMSO. Then, conventional EM grid vitrification was compared with simplified EM grid vitrification. Simplified EM grid vitrification showed higher survival, fertilization and hatching rates than those of the slow freezing method (85.6% vs. 63.2%; 51.0% vs. 22.3%; 38.7% vs. 12.5%, p < 0.01, respectively). Moreover, simplified EM grid vitrification showed higher recovery, survival and fertilization rates than those of conventional EM grid vitrification (100% vs. 95.0%, p=0.024; 90.0% vs. 78.9%, p=0.033; 56.7% vs. 38.7%, p=0.021, respectively). Hatching rate tended to be higher for simplified EM grid vitrification compared to conventional EM grid vitrification (41.1% vs. 24.1%). In conclusion, simplified EM grid vitrification is a convenient and efficient method for cryopreservation of mouse mature oocytes, compared to conventional EM grid vitrification and slow freezing methods.
Pregnancy ; Oocytes/*cytology ; Mice, Inbred DBA ; Mice, Inbred C57BL ; Mice ; Male ; *Fertilization in Vitro ; Female ; Cryopreservation/*instrumentation/*methods ; Cell Survival ; Animals

Microencapsulated recombinant cells technology is a novel approach to tumors therapy. It is necessary to prepare a plenty of the microcapsules with better cell viability and higher endostatin production in order to bring this technology into the clinic. The in vitro culture and cryopreservation are very important parameters in the preparation of microencapsulated cells. In this work, we studied the effect of the in vitro culture and cryopreservation on microencapsulated recombinant cells growth and endostatin production and the effect of the in vitro culture on the cryopreservation of microencapsulated recombinant cells. The results showed that the time of in vitro culture potently affected microencapsulated recombinant CHO cells growth in vivo, endostatin production and the microcapsule stability. The microcapsule kept intact after 36 days of implantation when the in vitro culture time was under 4 days. The thawed microencapsulated recombinant CHO cells had better cell growth and higher endostatin production after 40 days of cryopreservation when the in vitro culture time was 4 days and 8 days. Therefore, the best in vitro culture time was 4 days according to the results of the in vivo culture and cryopreservation and the cryopreservation did not affect microencapsulated recombinant CHO cells growth in vivo, endostatin production and the microcapsule stability.
Animals ; CHO Cells ; Capsules ; Cell Culture Techniques ; Cell Proliferation ; Cells, Immobilized ; cytology ; metabolism ; Cricetinae ; Cricetulus ; Cryopreservation ; methods ; Endostatins ; biosynthesis ; Implants, Experimental ; Mice ; Technology, Pharmaceutical ; instrumentation ; methods ; Time Factors

OBJECTIVETo summarize the clinical effect of avulsed skin replantation of hand and foot via vacuum sealing drainage (VSD) combing low temperature technique.

METHODSFrom March 2012 to October 2013,13 cases with avulsed skin replantation of hand foot using combined technique included 8 males and 5 females with an average age of 32 years old ranging from 18 to 62 years. The time from injury to hospital was 1 to 4 hours (2.4 hour in average). The reasons of injury included machine injury in 7 cases and rolling over by cars in 6 cases. The parts of injuried involved finger in 2 cases,back of the hand in 5 cases and dorsum of foot in 6 cases. The area of avulsed skin was 5 cm x 6 cm to 12 cm x 16 cm,tendon and bone exposure was found in 4 cases. VSD was operated in all patients and the avulsed skin was refrigerated in the temperature of -4 °C or -80 °C. After 4 days, the skin stored in the -4 °C was replanted to the wounded place in 5 cases and in 3 cases the skin was planted to the donor site of flap. The skin stored in the -80 °C was replanted in 4 cases after 7 or 8 days, 1 case after 45 days.

RESULTSOf the 13 cases, 1 case of degloved injury from lower leg to dorsal foot,the replanted skin was necrosis completely; 1 case of degloving injury with fourth finger,the skin which replanted after 45 days survived approximately 30%,cured after skin-graft many times. In the other cases, the survival area of replanted skin was more than 85%, all cured after dressing. According to the standard of skin survival area evaluation by Jia et al, 11 cases showed excellent, 1 showed medium and 1 showed inferior. There were no complication about grafted skin rupture after the skin survived in 11 patients,after 4 to 22 months follow-up, the resiliency of grafted skin showed good. Sensation recovery was measured by BMRC standard: 3 cases of S3, 5 cases of S3, 3 cases of S2.

CONCLUSIONVSD combining lower temperature technique in skin replantation provides time and space for wound preparation and treatment plan for the patients who need second surgery, especially for the large area skin degloving,this method could utilize the degloved skin efficiently, decrease the donor site area, alleviate the pain and financial burden,reduce the scar formation of donor site and impediment.

Adolescent ; Adult ; Cryopreservation ; methods ; Drainage ; instrumentation ; methods ; Female ; Foot Injuries ; surgery ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Replantation ; Skin ; injuries ; Skin Transplantation ; Young Adult

OBJECTIVETo evaluate the functional outcome and the complications of allograft replacement in management of bone tumors.

METHODSBetween March 1992 and September 2002 164 patients underwent bone tumor resection and massive allograft reconstruction of bone defects. The length of the resected part ranged from 5 - 35 cm. The resections were classified as marginal or wide resections of the tumor on the basis of the Musculoskeletal Tumor Society staging system. Fresh-frozen allografts were employed as osteoarticular grafts (n = 95), hemi-condylar (n = 15), massive (n = 23), allograft-prosthesis composite (n = 12), intercalary grafts (n = 15) or hemi-pelvic grafts (n = 4). Most of the lesions were osteosarcoma and giant cell tumor of bone and located in proximal and distal femur, proximal tibia and humerus.

RESULTSAt a median follow-up of 47 months (range, 12 to 168 months) after the operation, 154 of the patients in the study were free of disease and 10 died of disease. Twenty-one (12.8%) patients had local recurrence and 38 (23.2%) nonunion. Late complications included 11 (6.7%) fractures of the allograft and 18 (11.0%) infections of the graft. Instability of the joint in the form of subluxation was noted in 13 (7.9%) patients. Ten extremities were amputated due to local recurrence or severe infection.

CONCLUSIONSAllografts can be used for reconstruction of bony defects after tumor resection. Allograft has nearly similar shape, strength, osteo-conduction and osteo-induction with host bone. Allograft implantation is a high complication reconstruction method, and the risk of recurrence increases when less surgical margin achieves.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; surgery ; Bone Substitutes ; Bone Transplantation ; instrumentation ; methods ; Child ; Cryopreservation ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Transplantation, Homologous ; Treatment Outcome

The aim of this study was to investigate the cryobiological characteristics of placental cord blood (PCB) cryopereserved by using BioArchive auto-preserved liquid nitrogen system (BioArchive system). After Hespan depletion of red blood cells, 5 ml mixture of DMSO and 10% Dextran 40 were added into 20 ml of enriched leukocyte. 53 PCB units were cryopreserved as following protocol: pre-freeze rate 10 degrees C/min, start freeze temperature -3 degrees C, end freeze temperature -10 degrees C to -15 degrees C, post freeze rate 2 degrees C/min, and end temperature -50 degrees C. After rapid thawing at 38 degrees C, the PCB were washed with 5% human serum albumin -10% Dextran 40 and centrifuged at 400 x g, 10 degrees C for 20 minutes. The results showed that the viability of nucleated cells post-thaw was (73.3 +/- 12.5)%, the CD34(+) cell content was (0.3 +/- 0.21)% for pre-freeze PCB and (0.45 +/- 0.36)% for post-t haw PCB. No significant difference for CFU-GM/-G/-GEMM counts was found between pre-freeze and post-thaw PCB. Thawed PCB contained in two compartments (20 ml and 5 ml) of a freezing bag showed similar viability and clonogenic capacity. Differential count of white blood cell was significantly changed. For post-thaw PCB, it was dramatically decreased for the percentage of granulocytes, and highly increased for the percentage of lymphocytes and monocytes. It was concluded that the condition for cryopreservation and thawing of PCB may be harmful to mature cells, and cells with large size, such as granulocyte, but suitable to lymphocyte and monocyte, especially for the cells with small size, such as CD34(+) cells.
Antigens, CD34 ; immunology ; Blood Preservation ; instrumentation ; methods ; Cell Survival ; Cryopreservation ; methods ; Female ; Fetal Blood ; cytology ; immunology ; Granulocytes ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Lymphocytes ; cytology ; Monocytes ; cytology ; Placenta ; cytology ; Time Factors