Autologous blood transfusion stimulates wound healing in diabetic mice through activation of the HIF-1α pathway by improving the blood preservation solution.

Transfusion of autologous blood is a timesaving, convenient, safe, and effective therapy from a clinical perspective, and often employed for the treatment of diabetic patients. Stabilization of HIF-1α has been widely reported to be a critical factor in the improvement of wound healing in diabetes. Therefore, our study reveals the roles of improved autologous blood in wound healing in diabetes, through autologous blood transfusion in a mouse model. Initially, BALB/c mice were subjected to streptozotocin for diabetic mouse model establishment. Diabetic mice were transfused with improved or standard autologous blood in perfusion culture system. Roles of improved autologous blood in mediating HIF-1α pathway were determined by measuring expression of VEGF, EGF, HIF-1α, and HSP-90. In order to assess the detailed regulatory mechanism of improved autologous blood in perspective of wound healing, cell proliferation, migration and cell cycle, fibroblasts isolated from diabetic mice were transfected with HIF-1α siRNA. Mice transfused with improved autologous blood exhibited increased levels of CD31 and α-SMA in skin tissues, and reduced TNF-α, IL-1ß, and IL-6 levels, indicating that improved autologous blood promoted wound healing ability and reduced the release of inflammatory factors. Diabetic mice transfused with improved autologous blood presented activated HIF-1α pathway. The survival rate, proliferation, and migration of fibroblasts were elevated via activation of the HIF-1α pathway. Taken together, improved blood preservation solution could enhance the oxygen carrying capacity of red blood cells and wound healing in mice with diabetes, which is achieved through regulation of HIF-1α pathway.

Remimazolam tosilate in upper gastrointestinal endoscopy: A multicenter, randomized, non-inferiority, phase III trial.

BACKGROUND AND AIM: Remimazolam tosilate (RT) is a new short-acting GABA(A) receptor agonist, having potential to be an effective option for procedural sedation. Here, we aimed to compare the efficacy and safety of RT with propofol in patients undergoing upper gastrointestinal endoscopy. METHODS: This positive-controlled, non-inferiority, phase III trial recruited patients at 17 centers, between September 2017 and November 2017. A total of 384 patients scheduled to undergo upper gastrointestinal endoscopy were randomly assigned to receive RT or propofol. Primary endpoint was the success rate of sedation. Adverse events (AEs) were recorded to evaluate safety. RESULTS: The success rate of sedation in the RT group was non-inferior to that in the propofol group (97.34% vs 100.00%; difference in rate -2.66%, 95% CI -4.96 to -0.36, meeting criteria for non-inferiority). Patients in the RT group had longer time to adequate sedation (P < 0.0001) but shorter time to fully alert (P < 0.0001) than that in the propofol group. The incidences of hypotension (13.04% vs 42.86%, P < 0.0001), treatment-related hypotension (0.54% vs 5.82%, P < 0.0001), and respiratory depression (1.09% vs 6.88%, P = 0.0064) were significantly lower in the RT group. AEs were reported in 74 (39.15%) patients in the RT group and 114 (60.32%) patients in the propofol group, with significant difference (P < 0.0001). CONCLUSION: This trial established non-inferior sedation success rate of RT compared with propofol. RT allows faster recovery from sedation compared with propofol. The safety profile is favorable and appears to be superior to propofol, indicating that it was feasible and well tolerated for patients.

Anatomical dimensions of the lumbar dural sac predict the sensory block level of continuous epidural analgesia during labor.

BACKGROUND: The anatomical dimensions of the lumbar dural sac determine the sensory block level of spinal anesthesia; however, whether they show the same predictive value during continuous epidural anesthesia (CEA) remains undetermined. We designed the present study to verify the efficacy of the anatomical dimensions of the lumbar dural sac in predicting the sensory block level during labor analgesia. METHODS: A total of 122 parturients with singleton pregnancies requesting labor analgesia were included in this study. The lumbar dural sac diameter (DSD), lumbar dural sac length (DSL), lumbar dural sac surface area (DSA), and lumbar dural sac volume (DSV) were measured with an ultrasound color Doppler diagnostic apparatus. CEA was performed at the L2-L3 interspace. After epidural cannulation, an electronic infusion pump containing 0.08% ropivacaine and sufentanil 0.4 µg/ml was connected. The sensory block level was determined with alcohol-soaked cotton, a cotton swab, and a pinprick. The analgesic efficacy of CEA was determined with a visual analog scale (VAS). The parturients were divided into two groups, "ideal analgesia" and "nonideal analgesia," and the groups were compared by t test. Pearson's correlation was performed to evaluate the association between the anatomical dimensions of the lumbar dural sac and sensory block level. Multiple linear regression analysis was used to create a model for predicting the sensory block level. RESULTS: In the ideal analgesia group, the height, DSL, DSA, DSV and DSD were significantly smaller, and the body mass index (BMI) was significantly larger (P < 0.05). In addition, the DSL demonstrated the strongest correlation with the peak level of pain block (r = - 0.816, P < 0.0001; Fig. 2A), temperature block (r = - 0.874, P < 0.0001; Fig. 3A) and tactile block (r = - 0.727, P < 0.0001; Fig. 4A). Finally, the multiple linear regression analysis revealed that DSL and BMI contributed to predicting the peak sensory block level. CONCLUSION: In conclusion, our study shows that the sensory block level of CEA is higher when the DSL, DSA, DSV and DSD of puerperae are lower. DSL and BMI can be treated as predictors of the peak sensory block level in CEA during labor analgesia.

NTN1 gene was risk to non-syndromic cleft lip only among Han Chinese population.

OBJECTIVE: Genome-wide association studies (GWAS) found NTN1, NOG and the region between CREBBP and ADCY9 were risks to non-syndromic cleft lip with or without cleft palate (NSCL/P). However, the association of single nucleotide polymorphisms (SNPs) in these genes with NSCL/P in Western China is unknown. SUBJECTS AND METHODS: We selected seven SNPs in NTN1, NOG and between CREBBP and ADCY9, and then performed transmission disequilibrium test (TDT), parent-of-origin effect and sliding window haplotype analysis to test the associations among 302 NSCL/P case-parent trios from Western Han Chinese. RESULTS: We found allele G at rs4791774 in NTN1 was significantly overtransmitted among non-syndromic cleft lip only (NSCLO) (p = 0.0067, OR = 1.79, 95% CI: 1.17-2.74); rs4791774 and rs9915089 tightly linked with each other among NSCL/P (D' = 0.87, r2  = 0.67) and haplotypes carrying the risk allele G at rs4791774 were always found to be overtransmitted from parents to cases. Motif analysis indicated that allele G at rs4791774 could greatly alter the affinity of Myc_disc7, so allele G at rs4791774 in NTN1 might modulate C-MYC transcription to participate in the aetiology of NSCLO. CONCLUSIONS: Our study suggested allele G at rs4791774 in NTN1 gene is risk of NSCLO, which could greatly increase the risk to have a baby with cleft.

Autologous blood transfusion augments impaired wound healing in diabetic mice by enhancing lncRNA H19 expression via the HIF-1α signaling pathway.

BACKGROUND: Impaired wound healing frequently occurs in diabetes mellitus (DM) and is implicated in impaired angiogenesis. Long non-coding RNA (lncRNA) H19 has been reported as being reduced in DM and played a critical role in inducing angiogenesis. Thus, we hypothesized that H19 may affect impaired wound healing in streptozotocin (STZ)-induced diabetic mice transfused with autologous blood preserved in standard preservative fluid or modified preservative fluid. METHODS: Fibroblasts in injured skin were isolated and cultured in vitro. After location of H19 in fibroblasts using fluorescence in situ hybridization (FISH), RNA-pull down, RNA immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP), Co immunoprecipitation (COIP) and dual luciferase reporter gene assay were used to verify the binding of H19 to HIF-1α. RESULTS: The modified preservative fluid preserved autologous blood increased the H19 expression in fibroblasts, and maintained better oxygen-carrying and oxygen release capacities as well as coagulation function. Furthermore, H19 promoted HIF-1α histone H3K4me3 methylation and increased HIF-1α expression by recruiting EZH2. H19 promoted fibroblast activation by activating HIF-1α signaling pathway in fibroblasts and enhanced wound healing in diabetic mice. CONCLUSIONS: Taken together, H19 accelerated fibroblast activation by recruiting EZH2-mediated histone methylation and modulating the HIF-1α signaling pathway, whereby augmenting the process of modified preservative fluid preserved autologous blood enhancing the postoperative wound healing in diabetic mice.

Activation of mammalian target of rapamycin induces lipid accumulation in the diaphragm of ventilated rats and hypoxia-treated C2C12 cells.

BACKGROUND: Our previous study demonstrated that ventilators increase diaphragmatic lipid accumulation in rabbits, but their cellular mechanism is poorly understood. Mammalian target of rapamycin (mTOR) plays an important role in atherosclerosis in rat vascular smooth muscle cells. The present study investigated the role of mTOR pathway activation in the diaphragmatic muscle of ventilated rats and hypoxia-induced C2C12 cells. MATERIALS AND METHODS: Male Sprague-Dawly rats were randomized into a control group (n = 8), controlled mechanical ventilation (CMV) group (n = 8), and CMV + Rapa group (n = 8). We evaluated the diaphragmatic contractility, lipid accumulation, and protein expression of the mTOR pathways. To explore the mechanism underlying ventilator-induced lipid accumulation, we observed protein expression of the mTOR and low-density lipoprotein receptor (LDLr) pathways in C2C12 cells under hypoxic and mTOR pathway inhibitor treatments. RESULTS: Compared with the control group, there was a significant decrease in the peak twitch and peak tetanic forces in the CMV group (384.24 ± 70.39 versus 496.33 ± 78.64 g/cm2, P < 0.05, and 869.24 ± 76.67 versus 1090.72 ± 118.91 g/cm2, P < 0.05, respectively). There was a significant increase in peak twitch and peak tetanic forces in the CMV + Rapa group compared with that in the CMV group (501.81 ± 23.15 versus 384.24 ± 70.39 g/cm2, P < 0.05, and 992.91 ± 88.99 versus 869.24 ± 76.67 g/cm2, P < 0.05, respectively). In the CMV group, there were significant increases in lipid accumulation (0.086 ± 0.009 versus 0.005 ± 0.002, P < 0.05) and expression of mTOR in diaphragmatic fibers compared with those in the control group (P < 0.05). Rapamycin prevented lipid accumulation in rats of the CMV + Rapa group compared with that in the CMV group rats (0.024 ± 0.004 versus 0.086 ± 0.009, P < 0.05). Compared with the CMV group, there was a significant decrease in the phosphorylated protein expression levels of mTOR in rats of the CMV + Rapa group (P < 0.05). Hypoxic conditions activated the mTOR and LDLr pathways in C2C12 cells, which were correlated with an increase in expression of the mTOR and LDLr pathways compared with the control group (P < 0.05). In C2C12 cells treated with hypoxia + rapamycin, activation of the mTOR and LDLr pathways was blocked compared with C2C12 cells treated with hypoxia (P < 0.05). CONCLUSIONS: These data suggest that CMV and hypoxia-induced activation of the mTOR pathway, resulting in lipid accumulation, and impaired the diaphragmatic contractile function. Therefore, pharmacologic agents that inhibit the mTOR pathway could potentially be useful for mitigating the diaphragmatic contractile dysfunction induced by mechanical ventilation.

Can thyromental distance be measured accurately?

Using the thyromental distance (TMD) measured based on the ultrasonographic location of the thyroid cartilage prominence as the criterion, we investigated the accuracy of TMD measurement by surface landmark identification of the thyroid cartilage prominence. Twenty-nine anesthetist resident volunteers were recruited, including 10 first-year residents, 9 second-year residents and 10 third-year residents. Each volunteer measured the other 28 volunteers' TMD. Then, the thyroid cartilage prominence of each volunteer was identified by ultrasonography of the junction of the vocal cord and thyroid cartilage, and the TMD was measured precisely. The error of the TMD measurement was determined by the minimal detectable difference (MDD) compared to the ultrasound measurement. A difference of greater than 5.4 mm between the TMD measured by volunteers and that based on ultrasound localization was defined as a measurement error. The measurement error rate of females' TMD was significantly higher than that of males' (50 vs 10%, P < 0.001). The error rates of anesthetist residents of first-year, second-year and third-year were 34, 27, and 31%, respectively, and were not significantly different. The error of TMD measurement by surface landmark identification is often, especially for women. More clinic experience don't improve it.

A Comparison of the Efficacy and Tolerability of the Treatments for Sciatica: A Network Meta-Analysis.

BACKGROUND: There remains a lack of a systematic summary of the efficacy and safety of various medicines for sciatica, and discrepancies among these exist. OBJECTIVE: The aim of this study is to comprehensively assess the efficacy of and tolerance to several medical options for the treatment of sciatica. METHODS: We performed a network meta-analysis and illustrated the results by the mean difference or odds ratio. The surface under the cumulative ranking curve (SUCRA) was used for indicating the preferable treatments. All data analyses and graphs were achieved via R 3.3.2 and Stata 13.0. RESULTS: The subcutaneous anti-tumor necrosis factor-α (anti-TNF-α) was superior to the epidural steroid + anesthetic in reducing lumbar pain in both acute + chronic sciatica patients and acute sciatica patients. The epidural steroid demonstrated a better ability regarding the Oswestry disability score (ODI) compared to the subcutaneous anti-TNF-α. In addition, for total pain relief, the use of nonsteroidal antiinflammatory drugs was inferior to the epidural steroid + anesthetic. The epidural anesthetic and epidural steroid + anesthetic both demonstrated superiority over the epidural steroid and intramuscular steroid. The intravenous anti-TNF-α ranked first in leg pain relief, while the subcutaneous anti-TNF-α ranked first in lumbar pain relief, and the epidural steroid ranked first in the ODI on the basis of SUCRA. In addition, their safety outcome (withdrawal) rankings were all medium to high. CONCLUSIONS: Intravenous and subcutaneous anti-TNF-α were identified as the optimal treatments for both acute + chronic sciatica patients and acute sciatica patients. In addition, the epidural steroid was also recommended as a good intervention due to its superiority in reducing ODI.

Can Mandibular Condylar Mobility Sonography Measurements Predict Difficult Laryngoscopy?

BACKGROUND: Limited mandibular condylar mobility plays an important role in difficult laryngoscopy. Indirect assessment methods, such as mouth opening, have been proven to be useful predictors of difficult laryngoscopy. Sonography is a new direct assessment method for the limited mandibular condylar mobility. However, whether this method could be used in predicting difficult laryngoscopy still remains unknown. This study aimed to observe its ability to predict difficult laryngoscopy. METHODS: Adult patients who were administered tracheal intubations for elective surgery under general anesthesia were enrolled in the study. Mandibular condylar mobility was assessed by sonography through condylar translation measurements. Beside mouth opening, other indirect variables that correlated with temporomandibular joint mobility, such as mandibular protrusion distance, upper lip bite test, and whether the condyle-tragus distance was <1 finger breadth, were also evaluated before anesthesia. The primary outcome was difficult laryngoscopy defined as the Cormack-Lehane level 3 or 4. RESULTS: A total of 484 patients were prospectively included, and difficult laryngoscopy was reported in 41 patients. The condylar translation prediction criterion for difficult laryngoscopy was ≤10 mm. The condylar translation was correlated with Cormack-Lehane level (Spearman correlation coefficient, -0.46; 99% confidence interval [CI], -0.55 to -0.36) and owned the highest area under the receiver operating characteristic curve (0.93; 99% CI, 0.90 to 0.96, compared with that of the other predictors, P < .001) with difficult laryngoscopy. The condylar translation ≤10 mm was with a considerable κ value (κ = 0.52; 99% CI, 0.37 to 0.67) to difficult laryngoscopy and proved to be an independent predictor by a multivariate logistic regression. CONCLUSIONS: Compared with indirect assessments, such as mouth opening and other parameters, mandibular condylar mobility, as assessed directly using sonography, was correlated with difficult laryngoscopy and demonstrated an independent and notably predictive property.

[Effects of ketamine, imipramine, and their combination on depression-like behaviors in Wistar Kyoto rats].

The aim of the present study was to investigate the effects of ketamine, imipramine, and ketamine plus imipramine on chronic depression-like behaviors of Wistar Kyoto (WKY) rats and underlying mechanism. Six-week-old Wistar rats were used as normal control. WKY rats, depression model animal, were injected intraperitoneally with ketamine (1 week, replaced with saline in 2(nd) week), imipramine (2 weeks), or ketamine in combination with imipramine. The depression-like behaviors were assessed by sucrose preference and forced swimming tests. Protein expressions of ß form of calcium/calmodulin-dependent protein kinase type II (ßCaMKII) and membrane fraction of glutamate receptor 1 (GluR1) were measured in corresponding brain tissue with Western blot. The results showed that, compared with Wistar rats, WKY rats exhibited decreased sucrose preference and extended immobility time. Ketamine alone did not affect depression-like behaviors of WKY, whereas imipramine or its combination with ketamine could significantly decrease the immobility time. Compared with Wistar rats, WKY rats showed up-regulated levels of ßCaMKII and membrane GluR1 protein expressions in habenula, and down-regulated level of membrane GluR1 protein expressions in the prefrontal cortex. Imipramine or its combination with ketamine could reverse these changes of protein expressions in WKY rats. There was no difference in reversing effect between imipramine and its combination with ketamine. Ketamine alone did not affect the ßCaMKII and membrane GluR1 protein expressions in the habenula, but increased membrane GluR1 protein expression in the prefrontal cortex of WKY rats. These results suggest 2-week imipramine treatment significantly improves depressive behavior in WKY rats; however, the addition of ketamine in the first week fails to enhance the effect of imipramine. The underlying mechanisms of imipramine's anti-depressive effect may be associated with the down-regulation of ßCaMKII and membrane GluR1 in the habenula, as well as the up-regulation of membrane GluR1 in the prefrontal cortex.

[Effect of stellate ganglion block on reconstruction of the left ventricle in spontaneously hypertensive rats].

OBJECTIVE: To determine the effect of stellate ganglion block on reconstruction of the left ventricle in spontaneously hypertensive rats (SHRs). METHODS: Thirty-two 10-week-old male SHRs were randomly assigned into 4 groups: a left stellate ganglion block group (group LS), a right stellate ganglion block group (group RS), a captopril group (group D) and a control group (group C). The arterial systolic blood pressure (SBP) was measured by ALC-NIBP measuring system. After 10 weeks, we observed the left ventricular mass index (LVMI), myocardial pathologic changes, and detected the endothelin (ET-1) and endothelial nitric oxide synthase (eNOS) level in the left ventricle by radioimmunoassay and the collagen protein level in the left ventricle by immunohistochemical method. RESULTS: Compared with group LS and group C, the LVMI in group RS was lowered most notably (P<0.05) and pathological changes were improved obviously. The expression of eNOS in group RS was significantly increased and ET-1 significantly decreased (P<0.05) compared with that in group C and group LS. The expression of type I collagen fibers in group RS was significantly lower and type III collagen fibers significantly higher (P<0.05) when compared with that in group C and LS. CONCLUSION: Right stellate ganglion block can not only decrease the arterial pressure but also reverse the reconstruction of the left ventricle.

[Effect of stellate ganglion block on cardiomyocyte apoptosis and expression of Bcl-2/Bax protein in spontaneously hypertensive rats].

OBJECTIVE: To investigate the relationship between apoptosis of myocardial cells in spontaneously hypertensive rats (SHRs) and the expressions of Bcl-2 and Bax protein, and the protective effect of stellate ganglion block on apoptosis of myocardial cells. METHODS: A total of 32 ten-week-old male SHRs were assigned randomly into 4 groups: a left stellate ganglion block group (group LS), a right stellate ganglion block group (group RS), a captopril group (group D) and a control group (group C). The arterial systolic blood pressure was measured by ALC-NIBP system. After 10 weeks, all rats were anaesthetized by 3% pentobarbital sodium, cardiomyocyte apoptosis index of left ventricle was assessed by TUNEL, and the localization of myocardium Bcl-2, Bax was investigated by immunohistochemistry. RESULTS: Compared with group LS and C, the apoptotic index decreased (P<0.05). SHR myocardial expression of Bcl-2 significantly increased (P<0.05), Bax expression significantly decreased (P<0.05) and Bcl-2/Bax was significantly higher (P<0. 05) in group RS. CONCLUSION: Bcl-2 and Bax play an important role in the apoptosis of myocardial cells in SHRs. Right stellate ganglion block can reduce the apoptosis of myocardial cells and reverse the reconstruction of the left ventricle in SHRs via regulation of apoptosis-related gene proteins.

[Effects of stellate ganglion block on blood pressure in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effects of stellate ganglion block (SGB) on blood pressure in spontaneously hypertensive rats(SHRs). METHODS: Thirty-two 10-week-old male spontaneously hypertensive rats(SHRs) were assigned randomly into four groups: left stellate ganglion block group(Group LS), right stellate ganglion block group(Group RS), captopril group(Group D) and control group(Group C). Arterial systolic blood pressure(SBP) was measured, and endothelin (ET-1) and endothelial nitric oxide synthase(eNOS) in blood vessels were detected by radioimmunoassay. RESULTS: Compared with baseline value, the blood pressure of Group LS gradually increased significantly (P<0.05 or P <0.01); however, the blood pressure of Group RS was stable(P >0.05) and increased only at week 2(P <0.05).The blood pressure of Group D decreased significantly at week 2 and week 4, and it remained stable compared with baseline value (P<0.05). The blood pressure of Group C gradually increased at weeks 2-10, compared with baseline values (P <0.01). Compared with Group LS and Group C, the expression of eNOS in blood vessels of Group RS significantly increased (P <0.05), and ET-1 decreased (P <0.05). CONCLUSION: The right stellate ganglion block can significantly lower blood pressure, down-regulate ET-1 and up-regulate eNOS protein expression.

4.8% sevoflurane induces activation of autophagy in human neuroblastoma SH-SY5Y cells by the AMPK/mTOR signaling pathway.

Prolonged sevoflurane exposure leads to neurotoxicity. Autophagy plays an important role in promoting cell survival in different conditions. However, the role and mechanism of autophagy in sevoflurane-induced neurotoxicity were not fully elucidated. We attempted to indicate whether sevoflurane could activate the AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR)-mediated autophagy to attenuate anesthetics-induced neuronal injury in this study. Sevoflurane treatment significantly decreased the cell viability and induced apoptosis of SH-SY5Y cells. The expression level of Bcl-2 decreased, while that of Bax remarkably increased. Meanwhile, autophagy was activated by sevoflurane exposure as evidenced by increased expression levels of autophagy-related proteins (LC3-II and Atg5), decreased expression level of autophagic substrate P62, and increased autophagosomes and autolysosomes. Further autophagosomes and fewer autolysosomes were observed in the presence of Bafilomycin A1, an autolysosomes degradation inhibitor, suggesting that sevoflurane induced autophagic flux rather than inhibiting degradation of autophagy. Activation of autophagy by rapamycin partly reversed the sevoflurane-decreased cell viability. In contrast, inhibition of autophagy by 3-Methyladenine (3-MA) or Atg5-targeted small interfering RNA (siRNA) aggravated the sevoflurane-induced neurotoxicity. Further examination revealed that sevoflurane-induced autophagy was mediated by the AMPK/mTOR signaling pathway, with increased p-AMPK expression and decreased p-mTOR expression. Collectively, these results indicated that sevoflurane activates autophagy by regulating the AMPK/mTOR signaling pathway, which is protective against sevoflurane-induced damage in SH-SY5Y cells. Our results may assist clinicians to develop further promising therapeutic strategies for the neurotoxicity induced by inhaled anesthetics.

Retraction Notice to: lncRNA MALAT1 Accelerates Wound Healing of Diabetic Mice Transfused with Modified Autologous Blood via the HIF-1α Signaling Pathway.

[This retracts the article DOI: 10.1016/j.omtn.2019.05.020.].

Effects of acute normovolemic hemodilution on perioperative coagulation and fibrinolysis in elderly patients undergoing hepatic carcinectomy.

OBJECTIVE: To observe the effects of acute normovolemic hemodilution (ANH) on coagulation function and fibrinolysis in elderly patients undergoing hepatic carcinectomy. METHODS: Thirty elderly patients (aged 60-70 years) with liver cancer (American Society of Anesthesiologists physical status I-II) scheduled for hepatic carcinectomy from February 2007 to February 2008 were randomly divided into ANH group (n = 15) and control group (n = 15). After tracheal intubation, patients in ANH group and control group were infused with 6% hydroxyethyl starch (HES) (130/0.4), and basic liquid containing 6% HES and routine Ringer's solution, respectively. In all the studied patients, blood samples were drawn at five different time points: before anesthesia induction (T1), 30 minutes after ANH (T2), 1 hour after start of operation (T3), immediately after operation (T4), and 24 hours after operation (T5). Then coagulation function, soluble fibrin monomer complex (SFMC), prothrombin fragment (F1+2), and platelet membrane glycoprotein (activated GPIIb/GPIIIa and P-selectin) were measured. RESULTS: The perioperative blood loss was not significantly different between the two groups (P > 0.05). The volume of allogeneic blood transfusion in ANH group was significantly smaller than that in control group (350.5 +/- 70.7 mL vs. 457.8 +/- 181.3 mL, P < 0.01). Compared with the data of T1, prothrombin time (PT) and activated partial thromboplastin time in both groups prolonged significantly after T3 (P < 0.05), but still within normal range. There were no significant changes in thrombin time and D-dimer between the two groups and between different time points in each group (all P > 0.05). SFMC and F1 + 2 increased in both groups, but without statistical significance. P-selectin expression on the platelet surface of ANH group was significantly lowered at T2 and T3 compared with the level at T1 (P < 0.05). Compared with control group, P-selectin was significantly lower in ANH group at T2-T5 (all P < 0.05). CONCLUSIONS: In elderly patients undergoing resection of liver cancer, ANH may not hamper fibrinolysis and coagulation function. It could therefore be safe to largely reduce allogeneic blood transfusion.

[Application of micellar electrokinetic capillary chromatography for identification, quantitative detection and unfolding analysis of interleukin-12].

OBJECTIVE: To establish a micellar electrokinetic capillary chromatography-based method for identification and quantitative detection of interleukin-12 (IL-12) and analysis of its unfolding process. METHODS: An uncoated fused-silica capillary (inner diameter 50 µm) with a total length of 48.5 cm (40 cm to the detector) was used for the experiment. The factors influencing the separation efficiency of IL-12 were analyzed, and a standard curve of IL-12 concentration was established. The mixture of IL-12 and anti-IL-12 antibody was incubated in a water bath at 38 ℃ for 40 min, and capillary electrophoresis was then performed under the same conditions. The results were compared with those of IL-12 and anti-IL-12 antibody to identify IL-12. IL-12 and dithiothreitol (DTT) were incubated at 60 ℃ in water bath for different lengths of times, and the unfolding process of IL-12 was analyzed based on electrophoresis results of IL-12 in different states. RESULTS: A micellar capillary electrophoresis on-line sweep method was established with 80 mmol/L borate (pH=9.3) containing 30 mmol/L sodium dodecyl sulfate (SDS) as the buffer solution. This system showed a good linear relationship between the peak area and the mass concentration of IL-12 with a linear correlation coefficient of 0.9991 within the linear range of 2 to 120 ng/L. As the incubation time of IL-12 and DTT prolonged, the disulfide bond of IL-12 gradually opened and resulted in distinct changes in the protein peak. CONCLUSIONS: This capillary electrophoresis-based method is simple and sensitive for IL-2 analysis and allows rapid detection of changes in IL-12 content in the setting of tumors and analysis of the possible causes.

Effects of Morphine on Interstitial Cells of Cajal in Rabbit Colon and Small Intestinal Transit: An Experimental Study.

OBJECTIVE: This study aims to investigate the effect of morphine with naloxone on intestinal peristalsis and the number of interstitial cells of Cajal (ICC) in colon tissues of rabbits. METHODS: Thirty rabbits were randomly divided into five groups (n=6, each group): saline control group (NS group), low concentration of morphine group (L group), medium concentration of morphine group (M group), high concentration of morphine group (H group), medium concentration of morphine and naloxone mixed with antagonist group (NM group). Rabbits in these five groups were administered with an epidural puncture tube and dorsal epidural analgesia pump, and were continuously infused for seven days. Fecal characteristics were observed, and the ink propulsion rate was calculated. The expression level of ICC C-kit protein in colon tissues was tested by western blot. RESULTS: The stool characteristics in the L, M and H groups were more severe than those in the NS and NM groups. Furthermore, the intestinal propulsion rate in the L, M and H groups was lower than that in the NS and NM groups. The C-kit mRNA and protein expression in the colon of rabbits were significantly lower in the L, M and H groups, when compared to the NS and NM groups. CONCLUSION: Naloxone blocked the mRNA and protein expression of C-kit, and improved intestinal motor function.