Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective To investigate the expressions of IL-18, IL-18 binding protein isoform a (IL-18BPa) and IL-18 receptor α (IL-18Rα) in blood CD4⁺ Th2 cells of patients with allergic rhinitis (AR) and the effects of allergens on their expressions. Methods Blood samples of AR patients and healthy control subjects (HCs) were collected. Peripheral blood mononuclear cells (PBMCs) and CD4⁺ T cells sorted by immunomagnetic beads were stimulated by crude extract of Artemisia sieversiana wild allergen (ASWE), Platanus pollen (PPE) and house dust mite extract (HDME). Flow cytometry was used to detect the expression of IL-18, IL-18BPa and IL-18Rα in CD4⁺ Th2 cells, and BioPlex was used to detect the level of plasma IL-4 and analyze its correlation with the proportion of IL-18⁺ Th2 cells. Results Compared with HCs, the proportion of IL-18⁺ cells was increased in Th2 cells of AR patients; MFI of IL-18 was increased, while that of IL-18Rα was decreased. Moreover, allergens induced IL-18 and IL-18Rα expression in sorted CD4⁺ Th2 cells of HCs and induced IL-18Rα in that of AR patients. Additionally, elevated plasma IL-4 level was found in AR patients, which was moderately correlated with the percentage of IL-18⁺ Th2 cells. Conclusion Allergens may be involved in the pathogenesis of AR by inducing expression of IL-18 in peripheral blood CD4⁺ Th2 cells.
Humans ; Th2 Cells ; Interleukin-18/metabolism* ; Up-Regulation ; Leukocytes, Mononuclear/metabolism* ; Interleukin-4/metabolism* ; Rhinitis, Allergic/metabolism* ; Allergens ; Cytokines/metabolism*

Objective:To investigate the changes of autonomic nervous active substances in myocardium of rats with acute high-level spinal cord injury.Methods:Twenty-four clean-level healthy adult male SD rats weighting 250-300 g for 8-10 weeks old were divided into control group ( n=6) and spinal cord injury group ( n=18) according to the random number table. The spinal cord injury group was subdivided at 4, 12 and 24 hours, with 6 rats at each time point. The high-level spinal cord injury model was established by the modified Allen′s weight-drop method, and the spinal cord was only exposed in control group. The postoperative performance and BBB score for limb movement were observed in each group. The myocardium of each group was resected and used to observe ultrastructure of myocardial cells under transmission electron microscope and detect protein and mRNA levels of tyrosine hydroxylase (TH), noradrenaline transporter (NET), acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) by Western blot and RT-PCR analysis. Results:Rats of control group showed normal limb motion after operation without significant change from the preoperation level, and mean BBB score was 21 points. Rats of spinal cord injury group showed significantly reduced activities and feeding, with flaccid paralysis of both lower limbs as well as no spontaneous excretion, and showed BBB score of 0 point at 4 hours and 12 hours after injury, which was increased slightly at 24 hours after injury, with the highest score for 1 point. The ultrastructure of myocardial cells showed no obvious abnormalities in control group, while different degrees of changes in spinal cord injury group. Compared with control group, Western blot analysis showed that protein levels of TH and NET were decreased, while AChE and ChAT were increased in spinal cord injury group ( P<0.05 or 0.01). Compared with control group, RT-PCR analysis showed that mRNA levels of TH and NET were decreased, while AChE and ChAT were increased in spinal cord injury group ( P<0.05 or 0.01). mRNA levels of TH and NET in spinal cord injury group at 24 hours after injury were significantly different from those at 4 hours and 12 hours after injury (all P<0.05). mRNA levels of ChAT in spinal cord injury group were statistically significant at 12 hours and 24 hours after injury from those at 4 hours after injury, with significant difference at 12 hours and 24 hours after injury (all P<0.05). Conclusion:Sympathetic nerve active substances TH and NET are down-regulated but vagal nerve active substances AChE and ChAT up-regulated in myocardium of rats with acute high-level spinal cord injury, which may be related to the relative excitation of the parasympathetic nerve blocking the sympathetic innervation of the higher center to the heart following high-level spinal cord injury.

Objective:To explore the risk factors of intensive care unit acquired weakness (ICUAW) in patients with sepsis, and to evaluate the predictive value of each risk factor for ICUAW.Methods:A case control study was conducted, 60 septic patients admitted to the intensive care unit (ICU) of Henan Provincial People's Hospital from October 20, 2020 to February 20, 2021 were enrolled. The patients were divided into two groups: sepsis ICUAW group and sepsis non-ICUAW group. The data of gender, age, body mass index (BMI), acute physiology and chronic health evaluationⅡ(APACHEⅡ) score, complications, mechanical ventilation, duration of ICUAW, length of stay in ICU, fasting blood glucose, blood lactic acid (Lac), procalcitonin (PCT), C-reactive protein (CRP), sequential organ failure assessment (SOFA) score, outcome, antimicrobial agent, glucocorticoid, sedatives and analgesics drugs and vasoactive drugs were collected. Risk factors were screened by univariate Logistic regression analysis, and odds ratio ( OR) was adjusted by multivariate binary logistic regression, P < 0.05 was considered as independent risk factors. Finally, the receiver operating characteristic curve (ROC curve) was drawn to analyze the predictive value of independent risk factors. Results:The APACHEⅡ score of the sepsis ICUAW group was significantly higher than that of the sepsis non-ICUAW group (23.05±8.17 vs. 15.33±4.89, P < 0.05), the total length of stay in the ICU was significantly longer than that of the sepsis non-ICUAW group (days: 15.1±9.2 vs. 8.5±3.4, P < 0.05), the improvement rate of patients was significantly lower than that of the sepsis non-ICUAW group [45.0% (9/20) vs. 95.0% (38/40), P < 0.05]. After univariate Logistic regression and multicollinearity test analysis, 7 factors including APACHEⅡ score, average SOFA score, blood lactic acid, proportion of mechanical ventilation, sedatives and analgesics drugs, type of antibiotics and type of vasoactive drugs were included in the binary Logistic regression model [ OR: 1.21, 2.05, 2.26, 0.21, 1.54, 2.07, 1.38, 95% confidence interval (95% CI): 1.09-1.35, 1.42-2.94, 1.12-4.57, 0.05-0.66, 1.03-2.29, 1.27-3.37, 0.96-2.00, all P < 0.05]. Hosmer-Lemchaw test P = 0.901, and the correct percentage of prediction was 85%, indicating good model fit. Multivariate binary Logistic regression analysis showed that APACHEⅡ score and average SOFA score were independent risk factors for the occurrence of ICUAW in septic patients (APACHEⅡscore: OR = 1.17, 95% CI was 1.004-1.376, P = 0.044; average SOFA score: OR = 1.86, 95% CI was 1.157-2.981, P = 0.01). ROC curve analysis showed that the mean value of APACHEⅡ score, average SOFA score and their combined detection had a certain predictive value for the occurrence of ICUAW in sepsis patients, areas under ROC curve (AUC) were 0.787, 0.881, 0.905, 95% CI was 0.646-0.928, 0.791-0.972, 0.828-0.982, all P < 0.05. When the cut-off value was 19.500, 6.225, 0.375, the sensitivity was 75%, 90%, 90%, and the specificity were 80%, 80%, 85%, respectively. Conclusion:APACHEⅡ score and average SOFA score can be used as independent risk factors for the occurrence of ICUAW in sepsis, and their combined predictive value is better than that of individual index.

Critical care medicine has developed rapidly and has become an indispensable comprehensive subject in clinical medicine at home and abroad.In recent years,the government has vigorously implemented the "Healthy China" strategy and strived to achieve a higher level of national health.The Henan Provincial People's Hospital has set up a network of interconnected "Wisdom · Critical Care Medicine Specialist League" to meet the major strategic needs of the country,and to play a role in attracting large hospitals to promote the sinking of quality medical resources and the improvement of grassroots service capabilities.Complementary advantages and resource sharing are conducive to achieve win-win cooperation and coordinated development between the third-grade class-A hospital and grassroots hospitals.

Objective To establish a pseudovirus system for phenotypic drug-resistance detection and provide a relatively cheap and easy method for drug-resistance testing. Methods EGFP gene was amplified from plasmid pSV-EGFP and then cloned to backbone plasmid pNL4-3.Luc. E-R-by double enzyme digestion;env gene was amplified from RNA isolated from HIV-1-infected persons and cloned to eukaryotic expression plasmid cells and EGFP or ENV expression. Pseudovirus was produced by co-transfection of two recombinant plasmids to 293t cells. Infection of pseudovirus was determined by co-cultured with TZM-b1 cells and immunofluorescent test. Results Two recombinant plasmids(mass ratio,pcDNA3.1-env:pNL4-3.EGFP.E-R-.=2:1)were co-transfected to 293t cells. Cultured supernatants containing pseudovirus were harvested at 48 h post-transfection. Fluorescence was observed in TZM-b1 cells after TZM-b1 cells were infected with pseudovirus at 48 h post-infection. Conclusion The recombinant pseudovirus carrying EGFP gene is constructed successfully and it could be used for phenotypic drug-resistance detection.

Objective To investigate the influence of“double low”technology(low concentration iodinated contrast agent and low-dose scan) combined with body mass index(BMI) on radiation dose and image quality of contrast-enhanced upper abdominal CT examination. Methods One hundred and twenty patients who received upper abdominal enhanced CT examination were randomly divided into 4 groups:group A1, the iodinated contrast agent iodixanol(270 mg/ml), BMI<18.5 kg/m2 and 80 kVp;group A2, the iodinated contrast agent iodixanol(270 mg/ml), 18.5 kg/m2≤BMI≤24.9 kg/m2 and 100 kVp; group B1, the iodinated contrast agent ioversol(320 mg/ml),<18.5 kg/m2 and 120 kVp; group B2, the iodinated contrast agent ioversol(320 mg/ml), 18.5 kg/m2≤BMI≤24.9 kg/m2 and 120 kVp. Image quality was subjectively scored, the objective parameters(noise, CT values of abdominal aorta and liver parenchyma, contrast noise ratio of abdominal aorta and liver parenchyma) were evaluated and radiation dose was recorded. The differences of the indexes between A1 and B1 groups, A2 and B2 groups were compared with Mann-Whitney U test and pared-samples t test. Results All CT images were good. No images with 4 scores were obtained. No significant difference was found between group A1 and B1, between group A2 and B2(P>0.05). There was no significant difference in contrast noise ratio of liver parenchyma(P>0.05), while significant differences existed in CT values of abdominal aorta and liver parenchyma, contrast noise ratio of abdominal aorta between group A1 and B1(P<0.05). Significant differences existed in the parameters above mentioned between group A2 and B2, respectively(P<0.05). Radiation dose was lower in group A1 than in group B1 and in group A2 than in group B2(P<0.05), respectively. Radiation dose was decreased by 40.1%(0.89/2.22) in group A1 than group B1 while radiation dose decreased by 56.9%(3.02/5.31) in group A2 than group B2. Conclusion According to BMI, the low concentration iodinated contrast agent and low-dose scan CT scanning technology could effectively reduce radiation dose and generate ideal images during the contrast-enhanced upper abdominal CT examination.

OBJECTIVETo investigate the distribution and proportion of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Autonomous Region.

METHODS152 HIV-1 patients were enrolled from 11 cities in Guangxi Autonomous Region from 2010 to 2012 by convenient sampling. Inclusion criterias were listed as the fdlowing: HIV-1 infection was confirmed by Western blot, HIV-1 viral load >1 000 copies/ml, > 18 year-old, and without any serious illnesses. 5 ml of peripheral blood samples were obtained from each patient. The viral RNA was isolated from plasma and used for amplification of full-length pol gene by nested RT-PCR. The amplified products were sequenced. After editing and modification, all sequences were characterized for preliminary subtyping by genotyping and confirmed with phylogenetic tree constructed by MEGA 5.03 software. The recombinant identification of 2 unknown recombinant strains was determined by RIP and jpHMM at GOBICS.

RESULTSAmong 152 patients, 137 full-length pol genes were successfully amplified and 127 HIV-1 subtypes were identified. The distribution and proportion of subtypes was summarized as the following 71 cases of CRF01_AE, accounting for 55.9% (71/127), 38 CRF08_BC, 29.9% (38/127), 13 CRF07_BC, 10.2% (13/127), and 3 B (B'), 2.4% (3/127), 2 unknown recombinant strains, 1.6% (2/127). In 11 cites of Guangxi Autonomous Region, subtype CRF01_AE was the dominant strain. Among heterosexual transmitted patients and drug abusers, the proportions of subtype CRF01_AE were 67.4% (58/86) and 34.1% (14/41), respectively. There was a significance different in the distribution of CRF01_AE in different routes of transmission (χ(2)=15.07, P<0.001). In age 21- 35, age 36- 60 and age>60 groups, the proportions of CRF01_AE was 43.6% (17/39), 57.6% (38/66), 77.3% (17/22), and CRF08_BC was 43.6% (17/39), 28.8% (19/66), 9.1% (2/22), respectively, the difference in proportions was significant(χ(2)=8.48, P= 0.014). The patterns of two unknown recombinant strains were found to be CRF01_AE/B (B') and CRF01_AE/C/B(B'), respectively.

CONCLUSIONCRF01_AE was the dominant HIV-1 subtype in Guangxi Autonomous Region from 2010 to 2012, with heterosexual transmission as its main spreading route. The two unknown recombinant strains in Guangxi Autonomous Region were reconstructed by subtype CRF01_AE and CRF_BC.

Blotting, Western ; China ; epidemiology ; Cities ; Drug Users ; Genes, pol ; Genotype ; HIV Infections ; epidemiology ; transmission ; virology ; HIV-1 ; genetics ; Humans ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; blood ; pol Gene Products, Human Immunodeficiency Virus ; genetics

As internal protein elements , inteins are self-excised from their host protein and catalyze ligation of exteins with a peptide bond .They are widely found in all the three domains of the biological kingdom , and in viral proteins .Intein-mediated protein splicing is a posttranslational autocatalytic process that does not require auxiliary enzymes or cofactors . Protein splicing involves four intramolecular reactions and a small number of key catalytic residues in the intein and exteins . The development of expressed protein ligation ( EPL ) and protein trans-splicing ( PTS ) is assisted by inteins and protein splicing.This review introduces the biosynthesis of backbone-cyclized peptides libraries , and describes the applications of cyclic polypeptides and their prospective applications in the future .

Objective To design and construct a new non-fusion soluble expression vector pTIG-mSUMO(small ubiq-uitin-related modifier) using the widely used solubility promoting protein SUMO and based on the translational coupling phenomenon in order to enable the non-fusion soluble expression of the broad-spectrum antiviral protein RA in Escherichia coli by pTIG-mSUMO.Methods The smt3 gene coding for SUMO protein was cloned from yeast genome DNA by PCR. After directed-site silent mutation to eliminate the EcoRⅠsite, the mutant mSUMO was inserted into pET-22b to obtain the translational coupling expression vector pTIG-mSUMO.The RA was subject to PCR amplification and cloned into the pTIG-mSUMO to obtain the expression plasmid pTIG-mSUMO/RA which was supposed to direct the soluble expression of RA by the translational coupling with mSUMO.Results A translational coupling expression vector pTIG-mSUMO which could di-rect/drive the SUMO and heterogonous protein non-fusion expression simultaneously was designed and constructed.The Western blotting result indicated that pTIG-mSUMO could direct the high-level expression of RA, around 40%of which was soluble.Conclusion A translational coupling expression vector pTIG-mSUMO is obtained.After coupling with SUMO, RA is highly expressed in E.coli and both the expression level and solubility are greatly improved.pTIG-mSUMO might contrib-ute to soluble expression of other proteins.