Objective To analyze the coincidence of the patients with capillary electrophoresis hemoglobin A 2 increase with defi-nitely diagnosed beta thalassaemia and to investigate its application value in the diagnosis of beta thalassaemia.Methods Two hundreds and sixty outpatients and inpatients with hemoglobin A 2 increase in our hospital from May 2014 to May 2015 were per-formed the genetic testing.Results Among 260 patients with hemoglobin A2 increase ,beta thalassemia gene mutations were detec-ted in 257 cases ,the coincidence rate reached 98.85% ,and the common 17 beta thalassemia gene mutations were not detected in the other 3 cases ,follow-up further detection of rare beta thalassaemia gene and beta globin gene sequencing was performed ,1 case of SEA-HPFH βdeletion type was found ,1 case was Taiwaneseβdeletion type and 1 case was Codon 89-93(-AGT GAG CTG CAC TG) heterozygous mutation ,it was verified that 3 cases of hemoglobin A2 increase without detecting 17 kinds of common beta thalassemia gene mutations were still beta chain mutation occurrence or big fragment deletion.At the same time ,among 257 speci-mens of beta thalassemia gene mutations ,42 cases were compound alpha thalassaemia ,accounting for 16.34% of beta thalassaemia. Conclusion Capillary electrophoresis hemoglobin A2 increase can provide a fast and accurate basis for beta thalassemia diagnosis , but which can not rule out the possibility of compoound alpha thalassaemia ,when the patient's hemoglobin A2 is increased ,alpha and beta thalassaemia genetic diagnosis should be simultaneously carried out.

Objective To observe the two different wetting fluid in airway humidification of patients with mechanical ventilation.Methods 40 patients with mechanical ventilation after cardiac surgery in our hospital from January to April,2009,were divided randomly into group A and group B,group A was given 0.45% sodium chloride solution 100ml plus ambroxol 15mg as airway humidification fluid,group B was given sterile water for injection100 ml plus ambroxol 15mg as airway humidification liquid.The amount of sputum aspiration,color,viscosity,the body temperature of patients,the lungs auscultation and chest X-ray were observed.Results No statistical difference was seen in sputum volume,color,viscosity,the body temperature of patients,lungs auscultation as well as chest X ray after mechanical ventilation for 4 hours,8 hours,16 hours,24 hours,1～2 d,3～5 d,6～7 d.No statistical difference was also seen in auscultation of the lung after mechanical ventilation for 4 hours,16 hours,24 hours,1～2 d,3～5 d,6～7 d.But auscultation of the lung in group B was better than that of group A after mechanical ventilation for 8 hours.Conclusions No sufficient fact can prove that different effect exists between 100 ml 0.45% sodium chloride solution plus ambroxol 15mg and 100 ml sterile water for injection plus ambroxol 15mg as airway humidification fluid during mechanical ventilation.

Aim To investigate the hypoglycemic pathway of terpenes from Cornus officinalis(TCF) from three aspects of insulin dependence, α-glucosidase inhibition, insulin sensitizing.Methods Insulin-deficient diabetes mellitus(DM) model was induced by tail vein injection of streptozotocin(STZ) into SD rats at the dose of 50mg·kg-1 body weight.Rats were randomly divided into seven groups: control group(CON), model group(Model), metformin group(Met) 0.1g·kg-1, shenqi jiangtang granules(Shenqi) group 1.0 g·kg-1, three dose groups of TCF: 0.10, 0.05, 0.025 g·kg-1.Body weight and blood glucose were measured every week.After four weeks, glycosylated hemoglobin (HbA1c), glycosylated serum protein (GSP) were determined.Normal ICR mice were divided into seven groups: CON, Model, Met group 0.2g·kg-1, acarbose group(Acar) 0.1 g·kg-1, Shenqi group 1.5g·kg-1, three dose groups of TCF: 0.20g·kg-1;0.10g·kg-1;0.05 g·kg-1.After 10 days of administration, intraperitoneal injections of glucose and gavage starch tolerance tests were employed.Normal SD rats were divided into six groups: CON, rosiglitazone group 0.02 g·kg-1, glipizide group 0.02 g·kg-1, three dose groups of TCF: 0.10, 0.05, 0.025 g·kg-1.After seven days of administration, intraperitoneal glucose tolerance test(IPGTT) was employed and levels of insulin was determined.Results (1)High dose of TCF significantly reduced the level of HbA1c(P<0.05), GSP(P<0.05) on STZ model rats;(2)TCF significantly improved the glucose tolerance and gavage starch tolerance in ICR mice(P<0.05);(3) High dose of TCF significantly reduced the blood glucose and serum insulin level.Conclusions TCF has obvious effects on inhibiting glucose absorb and promoting the use of glucose.It is able to exert hypoglycemic effect through non-insulin dependent pathway, whereas, whether it has the effects of α-glucosidase inhibition and insulin sensitization should be further validated.

Objective:In order to develop a comprehensive hospital debt risk model,it is imperative to conduct an analysis of the present debt risk landscape within public hospitals,so as to furnish hospitals with recommendations to avert potential debt risks.Methods:A debt risk model was constructed based on 7 common factors through the extraction and standardization of data from 25 indicators,using 600 public hospitals in a specific province as samples,followed by factor analysis.Results:The model's calculation of the comprehensive score of public hospitals aligns with professional explanations and provides a more accurate representation of the factors influencing the debt risk level of public hospitals,such as financial investment level,hospital type,hierarchy,and others.Conclusion:In conclusion,the model is efficient.It is imperative to adopt a comprehensive perspective on the matter of hospital debt and efficiently address the associated financial risks faced by public hospitals by fostering collaboration among multiple stakeholders and upholding fundamental principles.

Objective:To investigate the protective effect of hypothermic antegrade machine perfusion against canine ischemic brain injury.Methods:Thirteen beagle dogs were divided into the mild hypothermia with perfusion group ( n=6) and normothermia with perfusion group ( n=7) according to the random number table. The model of ischemic brain injury was established by neck transection. After 1 hour of ischemic circulatory arrest, the perfusion fluid based on autologous blood was continuously perfused through bilateral common carotid artery for 6 hours. The temperature of the perfusion fluid was set at 33 ℃ in the mild hypothermia with perfusion group and 37℃ in the normothermia with perfusion group, respectively. Blood oxygen saturation was recorded at 0, 1, 2, 3, 4, 5 and 6 hours after the beginning of perfusion to evaluate the perfusate oxygen level. The perfusate was collected, and the levels of Na +, K +, Ca 2+ and glucose as well as the pH value of the perfusate were detected in the two groups. At the end of perfusion, the parietal brain tissues of 1 dog from each group were collected to evaluate the water contents of brain tissues. Nissl staining was used to evaluate the morphological integrity of the pyramidal neurons in the frontal cortex and hippocampus. Neuronal nuclei antigen (NeuN) was used to evaluate the structural and morphological integrity of pyramidal neurons. Immunofluorescence glial fibrillary acidic protein (GFAP) and ionic calcium binding adaptor molecule 1 (Iba1) were used to evaluate the integrity and activity of astrocytes and microglia fragments. Results:At 0, 1, 2, 3, 4, 5 and 6 hours of perfusion, there was no significant difference in the blood oxygen saturation or Na + concentrations between the two groups (all P>0.05); the K + concentrations in the mild hypothermia with perfusion group were (4.57±0.12)mmol/L, (4.67±0.14)mmol/L, (4.27±0.12)mmol/L, (4.45±0.10)mmol/L, (6.60±0.15)mmol/L, (7.37±0.18)mmol/L and (9.03±0.16)mmol/L, respectively, which were significantly lower than those in the normothermia with perfusion group [(4.84±0.10)mmol/L, (5.31±0.13)mmol/L, (5.44±0.24)mmol/L, (5.70±0.18)mmol/L, (7.79±0.18)mmol/L, (10.44±0.40)mmol/L, (10.40±0.41)mmol/L] (all P<0.01). At 0, 1, 2 and 3 hours of perfusion, the Ca 2+ concentrations in the mild hypothermia with perfusion group were (0.72±0.15)mmol/L, (1.55±0.16)mmol/L, (1.62±0.15)mmol/L and (1.88±0.15)mmol/L, respectively, being significantly higher than those in the normothermia with perfusion group [(0.41±0.13)mmol/L, (0.99±0.12)mmol/L, (1.29±0.13)mmol/L, (1.57±0.11)mmol/L] (all P<0.01), and no significant differences were found at other time points (all P>0.05). At 0, 1 and 2 hours of perfusion, the glucose concentrations in the mild hypothermia with perfusion group were (5.75±0.19)mmol/L, (5.17±0.15)mmol/L and (4.72±0.15)mmol/L, respectively, being significantly higher than those in the normothermia with perfusion group [(5.30±0.22)mmol/L, (4.89±0.20)mmol/L, (4.30±0.17)mmol/L] (all P<0.01), with no significant differences found at other time points (all P>0.05). At 2, 3, 4, 5 and 6 hours of perfusion, the pH values of the mild hypothermia with perfusion group were 7.32±0.06, 7.25±0.02, 7.23±0.02, 7.24±0.02 and 7.24±0.02, respectively, being significantly higher than those in the normothermia with perfusion group (7.26±0.01, 7.21±0.01, 7.17±0.02, 7.15±0.02, 7.08±0.02) ( P<0.05 or 0.01), with no significant differences at other time points (all P>0.05). The water content of brain tissues in the mild hypothermia with perfusion group was (74.9±0.4)%, which was significantly lower than (79.9±0.9)% in the normothermia with perfusion group ( P<0.01). Nissl staining showed that the pyramidal neurons in prefrontal cortex and dentate gyrus had good integrity in the mild hypothermia with perfusion group. NeuN immunofluorescence staining showed that the morphology and structure of pyramidal neuron cells in the mild hypothermia with perfusion group were better with clearly visible axons than those in the normothermia with perfusion group, whereas the cytosol was full and swollen with scarce axons in the normothermia with perfusion group. GFAP and Iba1 immunofluorescence staining showed that more structurally intact glial cells, more abnormally active cells, thickener axons and better axon integrity in all directions were found in the mild hypothermia with perfusion group than those in the normothermia with perfusion group. Conclusion:Compared with normal temperature antegrade mechanical perfusion, the mild hypothermia antegrade mechanical perfusion can protect canine brain tissue and alleviate ischemic brain injury by maintaining stable energy and oxygen supply, balancing ion homeostasis and perfusion fluid pH value, reducing tissue edema, and maintaining low metabolism of pyramidal neurons, astrocytes and microglia.

Objective To investigate the effects of methylation inhibitor 5-Aza-2'-deoxycytidine (5-Aza-dC) on biological behavior of esophageal squamous carcinoma cell (ESCC) lines KYSE140 and KYSE150. Methods KYSE140 and KYSE150 cell lines were divided into the blank group, the control group and the experimental group. The cells in the blank group didn't do the treatment, and the cells in the control group were added to DMSO 2 μmol/L, while in the experimental group, cells were treated with different concentration (1, 2, 3 and 4 μmol/L) of 5-Aza-dC which affected respectively at different time (24, 48, 72 and 96 h). Cell proliferation was detected by using methyl thiazolyl tetrazolium (MTT) assay and the optimal drug concentration and time point were selected. Transwell assay was performed to detect the change of cell migration and invasion. Flow cytometry was used to observe the effects of drugs on cell apoptosis and cell cycle.The expression of PARP,Caspase-3,CCNB-1,and CCNE-1 were detected by Western blot. Results MTT result showed that the effective function time of 5-Aza-dC on KYSE140 and KYSE150 was 96 h at the concentration of 4 μmol/L. Under this condition, the cell ability of migration and invasion was decreased significantly. The migrated cell number of KYSE140 and KYSE150 respectively in the blank group, the control group and the experimental group was (193.3±8.6), (184.0±10.4), (61.7±7.1) and (112.0±6.4), (101.3± 7.9), (26.3±5.7). The invasive cell number was (47.3±7.3), (38.7±5.1), (8.0±3.9) and (83.3±6.8), (74.7±5.7), (21.0±2.7), respectively. The difference was statistically significant (P <0.05). Flow cytometry revealed that 5-Aza-dC increased the apoptosis of KYSE140 and KYSE150. The apoptosis rate of the blank group, the control group and the experimental group was (2.8±0.3) %, (11.2±0.7) %, (18.6±0.6) % for KYSE140 and (2.7±0.4)%,(9.8±0.4)%,(17.7±0.5)% for KYSE150.Compared with the other two groups,the cell number of G2/M phase in the experimental group was increased remarkably (P < 0.05). PARP and Caspase-3 were sheared evidently and the protein expression of CCNB-1 was up-regulated while the expression of CCNE-1 was down-regulated in the experimental group. Conclusion 5-Aza-dC can inhibit the proliferation and promote apoptosis of ESCC cells.

Objective To evaluate the preservation effect of normothermic mechanical perfusion with red blood cells from humanized genetically modified pig on severed human limbs. Methods Severed human limbs were perfused with red blood cells from humanized genetically modified pigs for 6 h. Perfusion solution was taken every hour to measure the oxygen partial pressure, Na⁺, K⁺, Ca²⁺, pH value, glucose, lactic acid and creatine kinase levels. Superficial flexor muscle was sampled to detect the changes of tumor necrosis factor (TNF)-α, interleukin (IL)-2 and IL-1 levels. At 0 and 6 h after perfusion, the superficial flexor muscles of the forearm were taken for pathological examination. Intercellular space and glycogen consumption of skeletal muscles were observed. An appropriate amount of forearm vessels was collected every 2 h to detect the apoptosis of vascular endothelial cells. X-ray angiography was performed before perfusion and 6 h after perfusion to observe the filling degree of finger-tip peripheral vessels. Results The oxygen partial pressure was observed in the normal range throughout the perfusion. Na⁺ concentration peaked at 1 h, reaching 138.7 mmol/L, and then fluctuated within the normal range. K⁺ level peaked at 2 h up to 6.08 mmol/L, then decreased and fluctuated within the normal range. Ca²⁺ concentration reached the peak at 4 h, up to 1.03 mmol/L. Glucose level was gradually decreased at the beginning of perfusion, reaching the lowest value of 17.7 mmol/L at 2 h after perfusion, and then maintained a dynamic balance. The pH value was decreased to 7.28 at 6 h after perfusion. The lactic acid level was increased to 9.6 mmol/L at 1 h after perfusion, and then gradually decreased. The creatine kinase level was increased at the start of perfusion, reached the peak at 2 h up to 20 030 U/L, then decreased and remained stable at the end of perfusion. At the end of perfusion, the morphology of muscle fibers was normal, the gap among muscle fibers was expanded slightly, and the glycogen of skeletal muscles was not significantly accumulated. At 0 h perfusion, the number of apoptotic cells in vascular endothelial cells was large, which was declined at 6 h perfusion. Evident vascular filling was observed at 0 h, and the filling degree of some finger-tip vessels was decreased at 6 h. Conclusions Normothermic mechanical perfusion of severed human limbs with red blood cells from humanized genetically modified pigs may continuously and stably supply energy and oxygen, adjust the ion pH balance of perfusion solution, maintain normal cellular metabolism and exerts certain protective effect upon severed human limbs.

Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

OBJECTIVE To investigate the effect of dioscin on renal injury in septic rats and its possible mechanism. METHODS The septic rat model was induced by using cecal ligation and puncture. Sixty model rats were randomly divided into model group （0.5% sodium carboxymethyl cellulose solution）， dioscin low-dose， medium-dose and high-dose groups （30， 60， 120 mg/kg） and dexamethasone group （positive control， 10 mg/kg）， with 12 rats per group； another 12 rats were selected as the sham operation group （0.5% sodium carboxymethyl cellulose solution）. After 15 minutes of modeling， rats in each group were injected with medicine/0.5% sodium carboxymethyl cellulose solution via the tail vein. Twenty-four hours after administration， the levels of creatinine （Cr）， blood urea nitrogen （BUN）， neutrophil gelatinase associated lipocalin （NGAL）， kidney injury molecule-1 （KIM- 1）， interleukin 6 （IL-6）， IL-1β and tumor necrosis factor-α （TNF-α） in serum and malondialdehyde （MDA） in renal tissue， superoxide dismutase （SOD） activity and the protein expressions of nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， NOD-like receptor protein 3 （NLRP3） were detected； renal histomorphology was observed. RESULTS Compared with model group， pathological injury of renal tissue was improved significantly in dioscin low-dose， medium-dose and high-dose groups； the levels of Cr， BUN， NGAL， KIM-1， IL-6， IL-1β and TNF-α in serum， MDA level and protein expression of NLRP3 in renal tissue were decreased significantly （P＜0.05）； SOD activity in renal tissue， protein expressions of Nrf2 and HO-1 were increased significantly （P＜0.05）， in a dose-dependent manner （P＜0.05）. The pathological damage of renal tissue in the dioscin high-dose group was similar to dexamethasone group， and there was no statistically significant difference in the levels of the above indicators （P＞0.05）. CONCLUSIONS Dioscin can activate the Nrf2/HO-1 signaling pathway to inhibit NLRP3 inflammasome， and realize the inhibition of inflammatory factors and oxidative stress， so as to protect the kidney injury in sepsis.

OBJECTIVE To investigate the effect of dioscin on renal injury in septic rats and its possible mechanism. METHODS The septic rat model was induced by using cecal ligation and puncture. Sixty model rats were randomly divided into model group （0.5% sodium carboxymethyl cellulose solution）， dioscin low-dose， medium-dose and high-dose groups （30， 60， 120 mg/kg） and dexamethasone group （positive control， 10 mg/kg）， with 12 rats per group； another 12 rats were selected as the sham operation group （0.5% sodium carboxymethyl cellulose solution）. After 15 minutes of modeling， rats in each group were injected with medicine/0.5% sodium carboxymethyl cellulose solution via the tail vein. Twenty-four hours after administration， the levels of creatinine （Cr）， blood urea nitrogen （BUN）， neutrophil gelatinase associated lipocalin （NGAL）， kidney injury molecule-1 （KIM- 1）， interleukin 6 （IL-6）， IL-1β and tumor necrosis factor-α （TNF-α） in serum and malondialdehyde （MDA） in renal tissue， superoxide dismutase （SOD） activity and the protein expressions of nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， NOD-like receptor protein 3 （NLRP3） were detected； renal histomorphology was observed. RESULTS Compared with model group， pathological injury of renal tissue was improved significantly in dioscin low-dose， medium-dose and high-dose groups； the levels of Cr， BUN， NGAL， KIM-1， IL-6， IL-1β and TNF-α in serum， MDA level and protein expression of NLRP3 in renal tissue were decreased significantly （P＜0.05）； SOD activity in renal tissue， protein expressions of Nrf2 and HO-1 were increased significantly （P＜0.05）， in a dose-dependent manner （P＜0.05）. The pathological damage of renal tissue in the dioscin high-dose group was similar to dexamethasone group， and there was no statistically significant difference in the levels of the above indicators （P＞0.05）. CONCLUSIONS Dioscin can activate the Nrf2/HO-1 signaling pathway to inhibit NLRP3 inflammasome， and realize the inhibition of inflammatory factors and oxidative stress， so as to protect the kidney injury in sepsis.