Expert consensus on the prevention, diagnosis and treatment of cold injury in China, 2020

Cold injury refers to local or systemic injury caused by a rapid, massive loss of body heat in a cold environment. The incidence of cold injury is high. However, the current situation regarding the diagnosis and treatment of cold injury in our country is not ideal. To standardize and improve the level of clinical diagnosis and treatment of cold injury in China, it is necessary to make a consensus that is practical and adapted to the conditions in China. We used the latest population-level epidemiological and clinical research data, combined with relevant literature from China and foreign countries. The consensus was developed by a joint committee of multidisciplinary experts. This expert consensus addresses the epidemiology, diagnosis, on-site emergency procedures, in-hospital treatment, and prevention of cold injury.

Dexamethasone prevents vascular damage in earlystage non-freezing cold injury of the sciatic nerve

Non-freezing cold injury is a prevalent cause of peripheral nerve damage, but its pathogenic mechanism is poorly understood, and treatment remains inadequate. Glucocorticoids have anti-inflammatory and lipid peroxidation-inhibiting properties. We therefore examined whether dexamethasone, a synthetic glucocorticoid compound, would alleviate early-stage non-freezing cold injury of the sciatic nerve. We established Wistar rat models of non-freezing cold injury by exposing the left sciatic nerve to cold（3–5°C） for 2 hours, then administered dexamethasone（3 mg/kg intraperitoneally） to half of the models. One day after injury, the concentration of Evans blue tracer in the injured sciatic nerve of rats that received dexamethasone was notably lower than that in the injured sciatic nerve of rats that did not receive dexamethasone; neither Evans blue dye nor capillary stenosis was observed in the endoneurium, but myelinated nerve fibers were markedly degenerated in the injured sciatic nerve of animals that received dexamethasone. After dexamethasone administration, however, endoneurial vasculopathy was markedly improved, although damage to the myelinated nerve fiber was not alleviated. These findings suggest that dexamethasone protects the blood-nerve barrier, but its benefit in non-freezing cold injury is limited to the vascular system.

BH3-only protein Bim is involved in myocardial injury induced by co-stress of ischemia and cold stress in rats

Objectives To investigate the effect of co-exposure of myocardial ischemia and cold stress on myocardial injury in rats and the relative mechanism.Methods Myocardial ischemia model was established by ligation of left coronary artery.SD rats were randomly allocated to 4 groups; sham+normal temperature(S group),sham+cold stress(SC group),myocardial ischemia+ normal temperature(Ⅰgroup), myocardial ischemia+cold stress(IC group).On the condition of 26℃,SC and IC groups were keeped in a 4℃artificial chamber for 8h(8;00-16:00) for 4 consecu- tive days.Car diac function was assessed by echocardiography;pathological change was analyzed by HE staining;myocardial infarct size was determined by TTC staining;Bim,Caspase-3 expression in myocardium was determined by western blotting.Results It was demonstrated that co-exposure of myocardial ischemia and cold stress could significantly make the cardiac muscle in abnormal shape,increase the infarct size and the expression of Bim and Caspase-3.Conclusions Co-exposure of myocardial ischemia and cold stress may aggravate the cardiac injury,pro- apoptosis protein Bim is involved.

Effects of Rainy or Snowy Day and Temperature Drop on Strawberry in Greenhouse and Countermeasures

From middle December, 2012 to early January, 2013, Jinan City was in- fluenced by cold airs and successive rainy and snowy days lasted for 13 d. Consequently, strawberry in greenhouse was seriously affected. In the research, the causes of strawberry cold injury were surveyed and analyzed, to provide references for scientific management for farmers.

How to protect liver graft with nitric oxide

Organ preservation and ischemia reperfusion injury associated with liver transplantation play an important role in the induction of graft injury. One of the earliest events associated with the reperfusion injury is endothelial cell dysfunction. It is generally accepted that endothelial nitric oxide synthase （e-NOS） is cell-pro- tective by mediating vasodilatation, whereas inducible nitric oxide synthase mediates liver graft injury after transplantation. We conducted a critical review of the literature evaluating the potential applications of regulating and promoting e-NOS activity in liver preservation and transplantation, showing the most current evidence to support the concept that enhanced bioavailability of NO derived from e-NOS is detrimental to ameliorate graft liver preservation, as well as preventing subse- quent graft reperfusion injury. This review deals mainly with the beneficial effects of promoting ＂endogenous＂ pathways for NO generation, via e-NOS inducer drugs in cold preservation solution, surgical strategies such as ischemic preconditioning, and alternative ＂exogenous＂ pathways that focus on the enrichment of cold storage liquid with NO donors. Finally, we also provide a basic bench-to-bed side summary of the liver physiology and cell signalling mechanisms that account for explaining the e-NOS protective effects in liver preservation and transplantation.

Hypothermic machine perfusion with metformin-University of Wisconsin solution for ex vivo preservation of standard and marginal liver grafts in a rat model

AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion(HMP).METHODS Eighteen young(4 mo old) and 18 aged(17 mo old)healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group(UWP), and UW solution with metformin perfusion group(MUWP). Aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), interleukin-18(IL-18), and tumor necrosis factor-alpha(TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase(e NOS) in liver sinusoidal endothelial cells were also examined.Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done. RESULTS AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively,significantly lower in the MUWP group than in the UWP group(P < 0.05), but no significant differences were found between the young and aged MUWP groups.Metformin increased the expression of AMPK and e NOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-e NOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.CONCLUSION The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.

Relevance of proteolysis and proteasome activation in fatty liver graft preservation: An Institut Georges Lopez-1 vs University of Wisconsin appraisal

To compare liver proteolysis and proteasome activation in steatotic liver grafts conserved in University of Wisconsin (UW) and Institut Georges Lopez-1 (IGL-1) solutions.METHODSFatty liver grafts from male obese Zücker rats were conserved in UW and IGL-1 solutions for 24 h at 4 °Cand subjected to “ex vivo” normo-thermic perfusion (2 h; 37 °C). Liver proteolysis in tissue specimens and perfusate was measured by reverse-phase high performance liquid chromatography. Total free amino acid release was correlated with the activation of the ubiquitin proteasome system (UPS: measured as chymotryptic-like activity and 20S and 19S proteasome), the prevention of liver injury (transaminases), mitochondrial injury (confocal microscopy) and inflammation markers (TNF 1 alpha, high mobility group box-1 (HGMB-1) and PPAR gamma), and liver apoptosis (TUNEL assay, cytochrome c and caspase 3).RESULTSProfiles of free AA (alanine, proline, leucine, isoleucine, methionine, lysine, ornithine, and threonine, among others) were similar for tissue and reperfusion effluent. In all cases, the IGL-1 solution showed a significantly higher prevention of proteolysis than UW (P < 0.05) after cold ischemia reperfusion. Livers conserved in IGL-1 presented more effective prevention of ATP-breakdown and more inhibition of UPS activity (measured as chymotryptic-like activity). In addition, the prevention of liver proteolysis and UPS activation correlated with the prevention of liver injury (AST/ALT) and mitochondrial damage (revealed by confocal microscopy findings) as well as with the prevention of inflammatory markers (TNF1alpha and HMGB) after reperfusion. In addition, the liver grafts preserved in IGL-1 showed a significant decrease in liver apoptosis, as shown by TUNEL assay and the reduction of cytochrome c, caspase 3 and P62 levels.CONCLUSIONOur comparison of these two preservation solutions suggests that IGL-1 helps to prevent ATP breakdown more effectively than UW and subsequently achieves a higher UPS inhibition and reduced liver proteolysis.

The Impact of Spring Disastrous Climate on Prunus sibirica var suavosperma Yield and Preventive Countermeasures

Using the data at standard ground observation field of Lingyuan Meteorological Bureau and comparative and parallel observation data during growth period of surrounding Prunus sibirica var suavosperma, and combining social investigation data about low-temperature cold injury loss of P. sibirica in recent years, it is found that low-temperature cold injury and abrupt change of temperature in spring are also important factors affecting P. sibirica yield and economic benefit. According to temperature indexes, meteorological department timely issues warning and forecast, and relevant departments and foresters take corresponding preventive measures, and strengthen cultivation technology and management of P. sibinca park. It avoids economic loss of P. sibirica caused by low-temperature cold injury, and obtains high and stable yield.

Effect of ulinastatin donor-pretreatment on liver graft during cold preservation in rats

Background Donor-pretreatment with ulinastatin may influence the liver graft during cold preservation. The aim of this research was to determine whether pretreatment of donor liver with Ulinastatin can attenuate cold preservation injury,and to explore the mechanism by which Ulinastatin affects the donor liver graft.Methods One hundred and forty-four Wistar rats were divided into the Ulinastatin treatment group （T group） pretreated with Ulinastatin 50 000 U/kg and control group （C group） treated with 0.9% normal saline via peritoneal injection prior to the anesthetization. After the abdominal cavity was opened and perfused with cold Ringer＇s lactate solution, the liver was harvested. The harvested liver was preserved in cold Ringer＇s lactate solution for 0, 2, 6, 24 hours, at which time the liver tissue was sampled for determination of dry weight and wet weight, Na＋-K＋-ATPase and Ca2＋-ATPase activity, lactic acid dehydrogenase （LDH） activity, lactic acid and malondialdehyde levels. Light microscopy and electron microscopy were used to observe liver morphology. The liver cold-preservation solution was taken for measurement of aspartate aminotransferase （AST） and alanine transaminase （ALT） levels. Correlation between ATPase activity and lactic acid level was analyzed by SPSS 13.0 for Windows.Results The morphology in the T group had improved cell boundaries vs. The C group at each time point. Dry weight to wet weight in the T group was lower than in the C group at 6 hours （P 〈0.05）, but the difference was not significant at 24 hours. ALT levels in the T group were lower than that in the C group at 6 hours （P 〈0.05） and 24 hours （P 〈0.01）. AST levels in the T group were lower than those in the C group at 2 hours （P〈0.05）, 6 hours （P 〈0.01） and 24 hours （P 〈0.01）.Na＋-K＋-ATPase activity in the T group was higher than in the C group and the mean difference between two groups was significant at 0 hour （P 〈0.05） and 2 hours （P 〈0.05）. Ca2＋-ATPase activity in the T group was higher than in the C group with the mean difference between two groups significant at 2 hours （P 〈0.05）. The T group had increased lactic acid levels at 0 hour （P 〈0.01） and 2 hours （P 〈0.05） compared with the C group, but there was no influence on the LDH activity at the same time. There were no obvious differences in the levels of malondialdehyde between the two groups at any time point. A linear correlation between Na＋-K＋-ATPase activity and lactic acid levels （r=0.295, P 〈0.05） was found.Conclusions Donor-pretreatment with ulinastatin may protect the cells in a liver graft from ischemia injury during cold preservation; the mechanism may be due to its promotion for cell glycolysis and its preservation of ATPase activity.

Detection and assessment of nitrogen effect on cold tolerance for tea by hyperspectral reflectance with PLSR, PCR, and LM models

Knowledge of nutrients effect on freezing tolerance is vital for protecting tea plants against cold injury(CI).Freezing injury treatments on tea leaves with different low temperature and nitrogen(N)concentration was evaluated by hyperspectral imaging based on the reflectance as potential analytical tool.Subsequently,quantitative evaluation of the CI was improved by comparing analyzed results using partial least squares regression(PLSR),principle component regression(PCR)and linear model(LM)models.Reflectance of the CI was obtained by hyperspectral imaging system in the band between 871 and 1766 nm.The results showed that average reflectance increases with the rise of N concentration.A substantial portion of the leaf from the plant with 100%N dosage had the darkest image and resulted in the highest reflectance because the N weakened the negative effect of freezing stress.Out of the five spectral domains tested,the best predictive accuracy for the CI of the tea leaf was achieved by PCR(R^(2)=0.9971,RMSE=0.0609)in 1410–1766 nm wavelength,followed by LM(R^(2)=0.9999,RMSE=0.0805)in 871–1000 nm.The whole interval had the worse prediction accuracy which could be caused by large variations in the data for a specific treatment and high absorption peak occurring around 1450 nm in the reflectance curve.The averages,R^(2) and RMSE for all the three statistical models showed that the worse prediction accuracy occurred in 1410–1766 nm,followed by the whole interval 871–1766 nm.The prediction accuracy was low which could be due to the strong water absorption peaks that appeared in both range of wavelengths.The models without absorption bands had improved correlation coefficient and reduced RMSE values between the measured and the predicted CI(R^(2)=0.9676;RMSE=0.3067)compared to the model developed with the reflectance values in the entire waveband(R^(2)=0.9504;RMSE=0.6629).The reflectance values where absorption bands occurred had detrimental effect on the model performance,which resulted in the lower correlation values and larger errors.This study has demonstrated that the wavelength at which absorption bands occur can influence model performance significantly which is a step towards real-time implementation of the technique.

Study on the inflammatory cytokines after heterotopic transplantation of isolated mouse heart preserved in a high-pressured mixed gas chamber

Background The maintenance of heart viability is important for heart transplantation. Currently, heart preservation is limited to 6 hours of cold ischemic storage. This study explored a new heart preservation method under a high-pressured mixed gas chamber. Methods C57BL/6 male mice were used to establish the model of mice cervical heterotopic heart transplantation. Adult donor mice were randomly divided into three groups subjected to naive operation （Group A）, standard control （Group B） and experimental control （Group C）. The recipient mice were randomly divided into two groups subjected to standard control and experimental control. Group A： hearts were isolated; Group B： hearts were isolated and preserved in HTK solution at 4 ℃ for 8 h and transplanted; Group C： hearts were isolated and preserved in high pressured gas （PO2：3200 hPa ＋ PCO： 800 hPa = 4000 hPa） at 4 ℃ for 8h and transplanted. After transplantation, the state of re-beating and cardiac function were observed for Group B and C. At 24 h after transplantation, samples were collected for HE staining, cardiac cell apoptosis detection by Tunnel staining and analysis of tumor necrosis factor α （TNF-α）, interleukin-1β （IL-1β）, interleukin-6 （IL-6） and interleukin-10 （IL-10） by reverse transcriotion-polymerase chain reaction （RT-PCR）. Results In group C, 15 transplanted hearts were re-beat, while only 6 in Group B. The re-beating rate in Group C was significantly higher than Group B [75.0%（15/20） vs. 30.0%（6/20） ,P = 0.01]. The time of re-beating was significantly different between Group B, and C [（352.35 ± 61.07）s vs. （207.85 ± 71.24） s, P 〈 0.011. HE staining showed that pathologic changes such as ceil edema and inflammatory cell infiltration were more obvious in Group B and C than in Group A, but less obvious in Group C compared with Group B. Tunnel staining showed that Group B had more obvious apoptosis than Group A and C. RT-PCR results showed significant increase of TNF-α, IL-1β and IL-6 expression in Group B than Group C （P 〈 0.01, the expression of IL-10 was higher in Group C than that in Group B. Conclusion Highpressured mixed gas （PO2：3200 hPa ＋ PCO： 800 hPa = 4000 hPa） preservation can reduce cold ischemia and reperfusion injury of donor heart, therefore to maintain myocardial viability and prolong preservation time of donor heart.