Effects of miRNA-143 and the non-coding RNA MALAT1 on the pathogenesis and metastasis of HeLa cells.

Cervical cancer is a common female malignancy of global dimensions. MicroRNAs (miRNAs) play crucial roles in the development, differentiation, proliferation, and apoptosis of tumors. The non-coding RNA MALAT1 participates in various physiological processes that are important for proper functioning of the body. Here, we analyzed the expression of miRNA-143 and MALAT1 in HeLa cells to evaluate their roles in the occurrence and metastasis of cervical cancer. HeLa cells were divided into five groups depending on the treatment conditions, namely, transfected with miRNA-143, MALAT1, miRNA-143 inhibitor and the MALAT1 inhibitor, and the untreated control. Reverse transcription-polymerase chain reaction was used to analyze the expression of miRNA-143 and MALAT1, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to assess proliferation, the trans-well assay to study cell invasion and migration, and western blot to analyze the levels of E-cadherin and vimentin. The proliferation of HeLa cells increased upon treatment with the miRNA-143 inhibitor and decreased when treated with the MALAT1 inhibitor, compared to the proliferation of the groups that were transfected with miRNA-143 and MALAT1, respectively (P < 0.05). Thus, miRNA-143 decreased cell invasion and migration potency, downregulated vimentin and upregulated E-cadherin expression, while MALAT1 had the opposite effects. In conclusion, the low expression of miRNA-143 and high expression of MALAT1 in cervical cancer cells could possibly potentiate cell invasion/migration and alter the levels of vimentin and E-cadherin.

[Anatomy research of cervical laminoplasty with preservation of the posterior ligament complex.]

Objective: To evaluate the feasibility of cervical laminoplasty with preservation of the posterior ligament complex for enlarging the spinal canal. Methods: Six up-to-standard human corpse specimens were divided into two groups by simple randomization (start from C4 group, S4; start from C5 group, S5; 3 corpses in each group). Decompression operation of C3-C6 level was performed in a predetermined sequence by using the new procedure with preservation of the posterior ligament complex.The basic depth of spinal canal was measured with a depth gauge at fixed point after the right bone groove of single level was completed.The operation of the contralateral bone groove was continued, and then the spinal canal was measured again when the spinous process was pulled backward by using a tissue forceps until the ligament complex was just tight.Retreat value (RV) of vertebral lamina was obtained by calculating the difference between the two measurements.The earlier measured levels needed to be remeasured when the operation area increased by one level. Two independent sample and one-sample t test were used to analyze the measurement results. Results: RV of vertebral lamina was small after finishing the first level of the decompression operation [S4: (0.87±0.72) mm; S5: (1.83±0.29) mm], and the value reached its maximum after the completion of C3-C6 level.The overall average RVmaxs from C3 to C6 level were (2.37±0.52) mm, (4.27±0.78) mm, (3.73±0.93) mm and (2.16±0.77) mm, respectively.The overall average retreat rates (RR) were 17%±7%, 32%±9%, 29%±10% and 16%±6%, respectively. The overall average RVmax of C4 and C5 level reached or exceeded the decompression threshold value of 4 mm (t=0.839, -0.703, both P>0.05). The average RVmax of C4/C5 level was similar in the two groups (t=-1.204, 1.189, both P>0.05); however, the difference of average RVmax between C3 and C6 level was significant (t=-4.429, 4.196, both P<0.05). Conclusions: Cervical laminoplasty with preservation of the posterior ligament complex can enlarge the sagittal diameter of spinal canal and relieve the compression of spinal cord.In addition, RV of each level increases as the number of the operation level increases, and the ability of vertebral lamina to retreat is quite different from C3 to C6 level.The decompression effect in the middle of the operation area is better than that on the cranial and tail side.

Karyotype analysis with amniotic fluid in 12365 pregnant women with indications for genetic amniocentesis and strategies of prenatal diagnosis.

We explored the strategies of prenatal diagnosis by foetal karyotype analysis in pregnant women with indications for genetic amniocentesis. Karyotype analysis of amniotic fluid was performed on 12365 pregnant women with indications for genetic amniocentesis. The detection rates and distributions of abnormal karyotypes were observed in a variety of indications for genetic amniocentesis. The detection rates of abnormal karyotype were 57.4% in either a mother or father with chromosomal abnormality, 8.5% in the pregnant women with pathological ultrasound finding (PUF), 2.79% in the pregnant women with advanced age (35 years and over) and 2.23% in the women with abnormal maternal serum screening (MSS) tests. Foetal abnormal karyotype was found in 86 pregnant women with PUF; of the 86 pregnant women, 42 had trisomy 13, 18 or 21. Of the 12365 pregnant women, foetal abnormal karyotype was found in 428 (3.46%); of the 428 foetuses, only 154 had trisomy 13, 18 or 21. In the pregnant women with abnormal MSS, 111 foetuses had abnormal karyotype, but only 36 foetuses had trisomy 13, 18 or 21. We conclude that (1) ultrasound is an important approach to prevent the birth of foetuses with chromosomal disease. (2) Non-invasive prenatal DNA detection cannot completely replace invasive prenatal diagnosis and MSS. (3) The strategies of prenatal diagnosis: Genetic amniocentesis is strongly recommended for the pregnant women with indications for genetic amniocentesis. For pregnant women who refuse invasive prenatal diagnosis, non-invasive prenatal DNA detection is first performed. If the results of non-invasive prenatal DNA detection are negative, the pregnant women are followed up by ultrasound; if the results of non-invasive prenatal DNA detection are positive, the pregnant women should undergo invasive prenatal diagnosis.

Application analysis of cognitive nursing in thyroid surgery patients.

OBJECTIVE: The aim of the study was to explore the effect of cognitive nursing service on stress response in patients undergoing thyroid tumor surgery. PATIENTS AND METHODS: From January 2018 to June 2019, 60 patients with thyroid tumor were selected. The patients were divided into control group and experimental group with 30 cases each. Cognitive nursing was used in the observation group and routine nursing was used in the control group. RESULTS: The scores of SDS and SAS in the observation group were significantly lower than those in the control group (p < 0.05). The nursing satisfaction of the observation group was significantly higher than that of the control group (p < 0.05). The levels of systolic blood pressure, diastolic blood pressure, angiotensin Ⅱ and cortisol in cognitive nursing group were better than those in conventional group (p < 0.05). The incidence of pain and other complications in the cognitive nursing group was lower than that in the conventional group (p < 0.05). After nursing, the anxiety and depression in the study group was (34.1 ± 4.9) and (18.1 ± 5.1) respectively; the anxiety and depression of the control group were (42.8 ± 7.3) and (25.4 ± 5.9) respectively; the anxiety and depression in the study group were significantly lower than those in the control group (p < 0.05). The improvement of diastolic pressure, systolic pressure and heart rate in the study group was better than that in the control group (p < 0.05). CONCLUSIONS: The application of cognitive nursing can effectively improve the patients' cognition of disease and treatment, reduce the patients' bad mood, improve the treatment compliance, reduce the occurrence of stress reaction, and improve the safety of anesthesia and operation. Cognitive nursing intervention provides guarantee for patients' prognosis recovery, helps patients recover and discharge as soon as possible, and also has high application value, which is worth promoting and applying in major hospitals.

An umbrella review exploring the effect of radiotherapy for head and neck cancer patients on the frequency of jaws osteoradionecrosis.

OBJECTIVE: Efforts have been made to reduce epidemiological indicators of osteoradionecrosis in patients with head and neck cancer over recent years. This umbrella review aims to synthesize the information of the systematic reviews/meta-analyses investigating the effect of radiotherapy in patients with head and neck cancer on the frequency of osteoradionecrosis and to identify and analyze the gaps in current scientific literature. MATERIAL AND METHODS: A systematic review of systematic reviews with and without meta-analysis of intervention studies was conducted. Qualitative analysis of the reviews and their quality evaluation were performed. RESULTS: A total of 152 articles were obtained, and ten of them were selected for the final analysis, where six were systematic reviews and four were meta-analysis. According to the guide Assessing the Methodological Quality of Systematic Reviews (Amstar), eight articles included were of high quality and two of medium quality. These descriptive systematic reviews/meta-analyses included a total of 25 randomized clinical trials, showing that radiotherapy has positive effects on the frequency of osteoradionecrosis. Even though a reduction in the incidence of osteoradionecrosis was observed back in the history, in systematic reviews with meta-analysis, overall effect estimators were not significant. CONCLUSIONS: Differential findings are not enough to demonstrate that there is a significant reduction in the frequency of osteoradionecrosis in patients with head and neck cancer treated by radiation. Possible explanations are related to factors such as the type of studies analyzed, indicator of irradiated complication considered, and specific variables included in the analysis. Many systematic reviews did not address publication bias and did identify gaps in knowledge that require further clarification.

Dietary squalene supplementation alleviates diquat-induced oxidative stress and liver damage of broiler chickens.

The aim of this study was to explore the protective effects of squalene supplementation on growth performance, oxidative status, and liver function of diquat-challenged broilers. One hundred forty-four 1-day-old male Ross 308 broiler chicks were allocated to 3 groups, and each group consisted of 6 replicates of 8 birds each. The three groups were as follows: 1) nonchallenged broilers fed with a basal diet (control group), 2) diquat-challenged broilers fed a basal diet, and 3) diquat-challenged broilers fed with a basal diet supplemented with 1.0 g/kg of squalene. Broilers were intraperitoneally injected with 20 mg/mL of diquat solution at a dosage of 1 mL/kg of BW or an equivalent amount of saline at 20 d. Compared with the control group, weight gain and BW change rate during 24 h after injection were decreased by diquat challenge (P < 0.05), and the diquat-induced compromised growth performance was improved by squalene supplementation (P < 0.05). Diquat administration reduced plasma superoxide dismutase activity and increased malondialdehyde accumulation and glutathione peroxidase activity in both plasma and the liver (P < 0.05). In contrast, plasma glutathione peroxidase activity in diquat-challenged broilers was reduced by squalene supplementation (P < 0.05). The hepatic glutathione level was reduced by diquat administration (P < 0.05), whereas its level in plasma and the liver of diquat-challenged broilers was increased by squalene supplementation (P < 0.05). The relative liver weight of broilers was increased by diquat challenge (P < 0.05), with its value being intermediate in the squalene-supplemented group (P > 0.05). The plasma aminotransferase activities and total bilirubin concentration were increased by diquat challenge (P < 0.05), which were reduced by squalene supplementation (P < 0.05). The mRNA abundance of hepatic nuclear factor erythroid 2-related factor 2 (P < 0.05) was upregulated by diquat treatment, regardless of squalene supplementation. The mRNA abundance of hepatic glutathione peroxidase 1 and B-cell lymphoma/leukemia 2-associated X protein was upregulated by diquat challenge (P < 0.05), which was reversed by squalene administration (P < 0.05). Squalene increased NAD(P)H quinone dehydrogenase 1 mRNA abundance and decreased caspase 3 mRNA abundance in the liver of diquat-challenged broilers (P < 0.05). The results suggested that squalene can increase weight gain, improve oxidative status, and alleviate liver injury in diquat-challenged broilers.

Animal models of restless legs syndrome.

Restless legs syndrome (RLS) is a common disease with prevalence up to 10% in the general population. It is mostly a subjective condition, making animal models intrinsically difficult. General increased activity (urge to move) and limb movements consistent with periodic limb movements of sleep, seen in most patients with RLS, are currently our best behavioral markers. Our best understanding of human RLS demonstrates reduced central nervous system (CNS) iron stores and dysfunction of dopaminergic systems, which most likely involves the spinal cord. Based upon this knowledge, animal manipulations, including destruction of the A11 diencephalic-spinal tract and iron deprivation, have resulted in animal behavior consistent with RLS. Dopamine receptor type 3 knockout mice also show general increased activity. Pharmacologic blockade of dopamine receptors in rodents has also caused movements resembling periodic limb movements of sleep in older rodents but not in younger animals. More sophisticated animal modeling is needed to facilitate our understanding of RLS.

Evidence of a glycemic threshold for the development of cataracts in diabetic rats.

PURPOSE: This study was aimed to establish a possible correlation between the levels of plasma glucose and degree of lens opacification. Levels of glycation- and glycoxidation-products in different lens protein fractions were also estimated with an aim to determine the involvement of these products in lens opacification. METHODS: A wide range of hyperglycemia was induced by injecting different doses of streptozotocin to 1 month old rats and lenses were examined on the 75th, 90th and 150th day post-injection. Lens opacification was measured by Scheimpflug imaging and densitometry. Levels of plasma glucose and glycated hemoglobin were measured after overnight fasting. On 90th day, levels of Amadori products in lens water soluble (WS) fraction were measured by affinity chromatography. Similarly, advanced glycation end products (AGEs) in lens WS, urea soluble (US) and alkali soluble (AS) fractions were measured immunochemically using a monoclonal antibody against the major glycoxidation product, carboxymethyl lysine (CML). RESULTS: Different dosages of streptozotocin injection resulted in a broad range of plasma glucose levels in the rats which were grouped into three groups on the basis of their plasma glucose levels: mildly diabetic (< 170 mg/dl plasma glucose), moderately diabetic (190-350 mg/dl) and severely diabetic (> 400 mg/dl). On the 75th, 90th and 150th day post-injection, only the moderately and severely diabetic rats developed cataracts whereas lenses of the mildly diabetic rats remained clear. As seen on 90th day, levels of glycated hemoglobin and Amadori products in lens WS fraction increased significantly in the moderately and severely diabetic groups whereas in the mildly diabetic rats these levels remained more or less same as in the control group. Levels of CML in WS fractions remained unchanged between control rats and different diabetic groups, while US fractions showed a decrease in CML in both the moderately and severely diabetic groups compared to the controls and the mildly diabetic group. Interestingly, AS fractions contained the highest level of CML; the moderately and severely diabetic groups showed about 2-fold higher levels than the controls and the mildly diabetic group. CONCLUSIONS: This study strongly supports the existence of plasma glycemic threshold above which incidence of diabetic cataract formation increases exponentially. This threshold level seems to be at approximately 180 mg/dl or 10 mM plasma glucose. Significant increase in the levels of glycation and glycoxidation products mainly in cataract lenses suggests that glycation and glycation-mediated oxidation play an important role in the development of diabetic cataracts.

Epidemiologic study on the skin lesions of workers exposed to N,N'-methylene-bis-(2-amino-1,3,4-thiadiazole) and of children in the neighborhood of the factory.

The air in a factory producing N,N' methylene-bis-(2-amino-1,3,4-thiadiazole) (MATDA) and water in a nearby river were often polluted by the intermediate and finished product. The levels of MATDA in the air, soil, and river water were 0.1-0.8 mg/m3, 6-2 650 mg/kg, and 0.5-2 mg/kg, respectively. Seventy-nine workers engaged in MATDA production were observed; 44% of them suffered from contact dermatitis; and 72% showed changes of pigment in the skin (versus 2 and 16%, respectively, for the referents). All of the exposed workers had experienced dermatitis at some time. For 218 pupils of a primary school in the neighborhood of the factory, the incidence of hypopigmentation and pigmentation of the skin was 14 and 72%, respectively, but in the reference group the corresponding incidence rates were 0 and 20%. Pigment changes of workers' children increased with the duration of residence in the neighborhood of the factory. The results of the survey led to a change in the formulation of the product from a powder to an emulsion. Improved technology was also initiated which allowed most of the waste to be reused in the processing. After these control measures the dermatitis of the workers was greatly reduced.

[Changes in water-soluble, urea-soluble and membrane intrinsic proteins in human senile cataract].

Gel filtration of water-soluble protein shows a substantial increase in HM+ alpha-crystallin and a marked decrease in beta- and gamma-crystallins in cortical cataract. A decrease in beta 1-crystallin in cortical punctate opaque lenses is also striking. In nuclear cataractous lenses HM+ alpha- and beta-crystallin increase, while gamma-crystallin decreases. The urea-soluble protein from clear lenses contains mainly of alpha beta chain, whereas in cataractous lenses the relative amounts of the 28 and 23ku polypeptides (the components of beta-crystallin) increased markedly. In cataractous lenses the relative amount of membrane intrinsic proteins decreases slightly and it has little statistical meaning.

[Studies on constituents of Ilex chinensis Sims].

Several compounds characterised as protocatechuic acid, caffeic acid, syringin, rotundic acid and pedunculoside were isolated from the leaves of Ilex chinensis, a Chinese crude drug. A novel compound cyclohexanone pedunculosyl-3,23-O-acetal was identified and proposed as a pedunculoside derivative produced in the extracting procedure.

[Studies permeation of puerarin and its phospholipid complex through mouse skin in vitro].

OBJECTIVE: To investigate the skin permeation of puerarin and its phospholipid complex and compare the differences between their permeation rates and cumulative permeation amounts. METHOD: Performing a test of permeation through the mouse skin in vitro in an improved Franz diffusion cell. RESULT: The cumulative permeation amount of phospholipid complex was higher than that of puerarin in the first hour and then increased slowly, meanwhile the permeation rate of puerarin rose higher than that of the complex. CONCLUSION: Phospholipid complex of puerarin can permeate through the mouse skin rapidly up to a certain amount in a short time, then begins to release drug slowly and lastingly.

Changes of urea-soluble and intrinsic membrane proteins in rat lenses during the formation of galactose cataract.

Urea-soluble and intrinsic membrane proteins from normal and galactose cataractous rat lenses were analyzed by SDS-PAGE. During cataract formation, MP22 increased whereas MP26 decreased almost to nil and MP24 emerged. However, the relative amount of MP18 remained essentially unchanged. These results suggested a conversion of MP26 to MP22 during cataract formation. We also observed the changes in the relative abundance of the polypeptides of the urea-soluble fraction with cataractogenesis.

Regional differences in the polypeptide composition of pig lens intracellular matrix and plasma membrane.

Analysis of pig lens urea-soluble fractions (USF) by gel filtration and SDS-PAGE revealed that the cortical USF is characterized by a high content of noncrystallin bands of 42, 44, 52, 55, 100 and 200 kD. These bands are markedly decreased in the nuclear USF though the 44-kD band to a lesser extent. These results suggested that pig lens nucleus contains much less cytoskeletal proteins than the cortex. Analysis of pig lens membrane showed that MP26 and MP17 are the major intrinsic membrane proteins and the relative amount of alpha A and beta B1b chains attached to the membrane are more abundant in the nucleus than in the cortex.

Role of glycine 1 and lysine 2 in the glycation of bovine gamma B-crystallin. Site-directed mutagenesis of lysine to threonine.

To determine the role of Gly-1 and Lys-2 of bovine gamma B-crystallin in glycation and cross-linking, Lys-2 was changed to Thr by site-directed mutagenesis. A polymerase chain reaction was used to perform site-directed mutagenesis on the third codon (AAG-->ACG) of bovine gamma B-crystallin cDNA. The wild type and mutant cDNAs were cloned into pMON5743 and expressed in JM101 Escherichia coli cells, and the identity of gamma B-crystallin was confirmed by Western blotting after purification by cation exchange high performance liquid chromatography. Glycation of gamma B-crystallin by [14C]glucose was reduced significantly due to the mutation of Lys-2, supporting the view that Lys-2 is a major glycation site. Peptide mapping showed the presence of two major labeled peptides containing N-terminal sequences, and in the mutant these peptides had longer retention times and reduced radioactivity. Amino acid analysis, after glycation with [14C]glucose, revealed N-terminal glycine as the most predominant glycation site. Lys-2 was glycated slower than Gly-1 but faster than Lys-163. Glycation with DL-glyceraldehyde showed an important role for both Gly-1 and Lys-2 in the glycation-mediated gamma B-crystallin cross-linking.

The role of alpha- and epsilon-amino groups in the glycation-mediated cross-linking of gammaB-crystallin. Study of three site-directed mutants.

In the previous report we demonstrated that gammaB-crystallin is glycated predominantly at the N-terminal alpha-amino group (Casey, E. B., Zhao, H. R., and Abraham, E. C. (1995) J. Biol. Chem. 270, 20781-20786). To investigate the possible role of alpha- and epsilon-amino groups of gammaB-crystallin in glycation-mediated cross-linking, Lys-2 or Lys-163, or both, were mutated to threonine by site-directed mutagenesis in bovine gammaB-crystallin cDNA. Wild type and mutant gammaB-crystallins were expressed in Escherichia coli cells. Cross-linking studies were performed by incubating wild type and mutant gammaB-crystallins with glyceraldehyde, ribose, and galactose followed by SDS-polyacrylamide gel electrophoresis under reducing conditions. When both of the lysines of gammaB-crystallin were mutated to threonines (gammaB-K2T/K163T), the quantity of cross-linked products was greatly reduced, indicating that, despite the fact that the alpha-amino group is a major glycated site, epsilon-amino groups play a predominant role in cross-linking. Therefore, cross-linking ability depends not only upon the level of glycation but also upon which amino group is glycated. Steric hindrance may decrease the cross-linking ability of the alpha-amino group. Our results also show that Lys-2 and Lys-163 play almost equal roles in cross-linking of gammaB-crystallin. By incubating carbonic anhydrase, a protein with a blocked N terminus, and our novel "no lysine" gammaB (gammaB-K2T/K163T) with sugar, we were able to show for the first time that significant cross-linking occurs between lysines and non-lysine sites. The fact that pentosidine and imidazolysine, formed from ribose and methylglyoxal, respectively, were present in the cross-linked gammaB-crystallins revealed the existence of Lys-Arg and Lys-Lys cross-linking.

[Application of fetal bone in repairing the defected part of osteoma resection].

OBJECTIVE: To investigate the clinical results used fetal bone repairing the defect after osteoma resection. METHODS: In 8 hours after abortion, the fetal bones of four limbs were cut and kept in the normal saline with gentamycin. Then the graft was storaged under 6 degrees C-8 degrees C and the longest storage time was 48 hours. Since 1990, seven cases of osteoma had been cured with fetal bone graft. Among them, there were 5 males and 2 females, aged from 6 to 38 years, the bone defect volume ranged from 5.5 cm x 3.6 cm x 2.5 cm to 9.0 cm x 4.3 cm x 3.2 cm. During operation, the osteoma was resected radically, then the defected cavity was impacted with prepared fetal bone. RESULTS: After operation, the general condition was good, all the incision wound healed primarily. Sixteen weeks after operation, the defected cavity was repaired completely and the medullary cavity reopened. Follow-up for 8 months to 5 years, no recurrence or side-effect were observed. CONCLUSION: Because of the more rapid expansion and replacement effect, the stronger osteoinduction, and the lower immune response, the fetal bone is practical material for repairing the bone defect in osteoma resection.

[Study on the geographic distribution and serological typing of HFRS in Hebei Province].

Surveillance on the HFRS had been carried out during the period of 1986-1993. Rodent density ranged from 4.6%-12.2% with R.norvegicus exceeding 60% in all kinds of rodents. HV antigen carrier rate of R.norvegicus was 1.3%-7.5%, much higher than that among other rodents. A significantly positive correlation had been observed between the HFRS morbidity and the R.norvegicus density. Of 129 specimens from HFRS patients, 89% belonged to the type of SEOV. In the eastern and centre Parts of Hebei plain, HTNV occupied 1.6%. A total number of 9,609 HFRS patients had been identified during the period of 1980-1993. The plain area in Hebei province was recognized as the major epidemic area. The key measure for decreasing the HFRS morbidity was deratizationin in the residential area in January and February and immunization with HFRS inactivated vaccine.

Advanced glycation end products in human senile and diabetic cataractous lenses.

The authors prepared water-soluble (WSF), urea-soluble (USF), alkali-soluble (ASF), sonicated (SF), sonicated insoluble (SIF) and membrane (MF) fractions of lens proteins from human senile and diabetic cataractous lenses and age-matched clear lenses. Levels of advanced glycation end products (AGEs) including carboxymethyl lysine (CML), a glycoxidation product, were determined by both non-competitive and competitive enzyme-linked immunosorbent assay (ELISA). Distribution of AGEs in the various protein fractions was ascertained by SDS-PAGE and Western blotting. An overall increase in the levels of AGEs in diabetic cataractous lenses as compared to senile cataractous lenses and clear lenses has been observed. ASF and SF, both of which originated from the urea-insoluble fraction, showed the highest levels of AGEs. However, no clear-cut differences in CML levels were seen among clear lenses and senile and diabetic cataractous lenses. AGEs were found to be distributed mostly in the high molecular aggregates in all the fractions. These data suggest that AGEs contribute to protein aggregation and subsequent insolubilization.

Five year digital replantation series from the frigid zone of China.

A series of 208 digital replantations from the frigid zone within the People's Republic of China is presented. The extremely cold climate (often -20 degrees C to -30 degrees C) presents the problem of warming exposed amputated digits prior to replantation without causing further damage. An overall replantation survival rate of 94% is reported, and this includes 45 cases of multiple digit amputation. Longer periods of ischemia seem to be tolerated under these unusually frigid conditions. Problems encountered within the series and suggestions for successful outcomes are discussed.