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 influence of ferri ion on the growth, reproduction and energy metabolism of Leptospira interrogans ( L. interrogans), and to identify whether the LA_2690 and LA_3598 gene products functioned as ferritin and ferroxidase. Methods:Petroff-Hausser counting method was used to analyze the influence of ferri ion deficiency on the growth and reproduction of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain 56601 in EMJH medium. Spectrophotometry and Chemiluminescence method was used to detect whether ferri ion deficiency inhibited the synthesis of DNA and ATP in L. interrogans. The structures and functions of L. interrogans LA_2690 and LA_3598 genes were analyzed using bioinformatic softwares. Prokaryotic expression systems for LA_2690 and LA_3598 genes were established and the target proteins, rLep2690 and rLep3598, were extracted by Ni-NTA affinity chromatography. The ferroxidase activity of rLep2690 and rLep3598 was detected by spectrophotometry. After L. interrogans strain 56601 was used to infect human umbilical vein endothelial cells (HUVEC) and monocytes (THP-1), changes in the expression of LA_2690 and LA_3598 genes at transcription level were detected using real-time fluorescence quantitative RT-PCR (qRT-PCR). Results:In the ferri ion-absent EMJH medium, the growth and reproduction of L. interrogans as well as the DNA and ATP synthesis levels were significantly decreased ( P<0.05). The products of LA_2690 and LA_3598 genes were predicted as bacterioferritin (Bfr) and DNA-binding ferritin containing ferroxidase diiron centers, but the latter lacked the heme-binding site and ferroxidase core. The prokaryotic expression systems for LA_2690 and LA_3598 genes could efficiently express the target recombinant proteins. Both the purified rLep2690 and rLep3598 showed a single band on SDS-PAGE. The ferroxidase activity of rLep2690 and rLep3598 was 1 238.619 U/L and 60.052 U/L, respectively. The expression of LA_2690 and LA_3598 genes of L. interrogans at mRNA level was significantly elevated during infection of the two types of cells ( P<0.05). Conclusions:Ferri ion participates in the growth and reproduction of L. interrogans as well as the synthesis of DNA and ATP. LA_2690 and LA_3598 genes were essential for L. interrogans to infect cells, and the product of LA_2690 gene possessed a stronger ferroxidase activity.

Objective:To construct the expression vector of the fusion protein of human serum albumin (HSA) and thymopentin (TP5) and to express it in Pichia pastoris,and to elucidate the biological activity of fusion protein.Methods:The HSA-TP5 fusion gene was constructed by gene recombination and transfected into Pichia pastoris to construct the eukaryotic expression system of HSA-TP5.The recombinant eukaryotic expression plasmid of PPICZα-HSA-TP5 was obtained by agarose gel electrophoresis and purification reagent.The two step fermentation method was used to ferment gene engineering bacteria of HSA-TP5 in high density,and the fermentation supernatant protein was precipitated and concentrated;the purified fusion protein was obtained by cation exchange chromatography and hydrophobic chromatography and analyzed by SDS-polyacrylamide gel electrophoresis.The effect of the fusion protein on the proliferation of lymphocytes was detected by MTT assay.Results:The HSA target gene fragment with length of 1 845 bp was achieved by PCR method.The HSA-TP5-pPICZαC fusion plasmid was identified by restriction endonuclease digestion,and the fragment length was 707 bp.The sequence analysis showed that the HSA and TP5 sequences of the target genes were identical with the gene sequences reported in GenBank and were fused by forward fusion.PCR method confirmed that the eukaryotic recombinant plasmid PPICZ αC-HSA-TP5 was integrated into the yeast genome,and compared with control group,the target gene PCR product length was found to be 1 860 bp.SDS-PAGE analysis showed that the expression level of HSA-TP5 fusion protein was gradually increased with the induction time within 72 h.HSA-TP5 fusion protein was purified by cation exchange chromatography and AKTA multifunctional protein purification system.The MTT assay results showed that HSA-TP5 fusion protein was consistent with TP5 protein in promoting lymphocyte proliferation activity.Conclusion:HSA-TP5 fusion protein can be obtained by constructing the eukaryotic expression system of Pichia pastoris and owns the biological activity.

This study aimed to validate the high yield and soluble expression of proteins carrying the transactivator of transcription (Tat) peptide tag, and further explored the potential mechanism by which the Tat tag increases expression. Escherichia coli superoxide dismutase (SOD) proteins, including SodA, SodB and SodC, were selected for analysis. As expected, the yields and the solubility of Tat-tagged proteins were higher than those of Tat-free proteins, and similar results were observed for the total SOD enzyme activity. Bacterial cells that overexpressed Tat-tagged proteins exhibited increased anti-paraquat activity compared with those expressing Tat-free proteins that manifested as SodA>SodC>SodB. When compared with an MG1655 wild-type strain, the growth of a ΔSodA mutant strain was found to be inhibited after paraquat treatment; the growth of ΔSodB and ΔSodC mutant strains was also slightly inhibited. The mRNA transcript level of genes encoding Tat-tagged proteins was higher than that of genes encoding Tat-free proteins. Furthermore, the α-helix and turn of Tat-tagged proteins were higher than those of Tat-free proteins, but the β-sheet and random coil content was lower. These results indicated that the incorporation of the Tat core peptide as a significant basic membrane transduction peptide in fusion proteins could increase mRNA transcripts and promote the high yield and soluble expression of heterologous proteins in E. coli.
Escherichia coli* ; Escherichia* ; HIV-1* ; Membranes ; Paraquat ; RNA, Messenger* ; Solubility ; Superoxide Dismutase ; Superoxides* ; Trans-Activators

Objective To study the clinical features and follow-up of newborns with severe hypoxic-ischemic encephalopathy ( HIE) , and to provide the basis for rational diagnosis, treatment and follow-up.Methods Clinical data of cases of HIE from the Neonatal Department of our Hospital from January 2011 to October 2014 were studied retrospectively. The data of general information, laboratory examination, treatment, outcome, follow-up and prognosis of the patients were collected. Multivariate logistic regression analysis was used to study the influential factors of the prognosis of HIE.Results A total of 123 infants with sever HIE were enrolled in our study. In addition to general therapy, 6 cases were treated with mild hypothermia, and 21 cases were treated with high pressure oxygen. 60 cases improved our treatment, 55 cases had withdrawal treatment with parental consent, and 8 cases died. Single factor analysis showed that 5 minutes Apgar score, convulsions, coma, pH, BE, organ injury, and mild hypothermia treatment were the risk factors that affect the prognosis of severe HIE. Multiple factors analysis showed that 5 min Apgar score <3 points ( OR=4. 071 ,95℅CI 1. 309-15. 613 ) and BE≤-10 mmol/L ( OR=36. 810, 95℅CI 5. 913-41. 119) were independent risk factors of prognosis of severe HIE ( P<0. 05). Hospitalization within the first 72 hours of life ( OR=0. 096, 95℅CI 0. 096-0. 353) was a protective factor of severe HIE. Multiorgan injury ( mainly the injury of brain, lung and heart) and electrolyte imbalance ( mainly hypocalcemia and hyponatremia ) were common complications of serve HIE. In the follow-up of these patients, 33 cases were loss in follow up, and 49 cases died (8 cases died during hospitalization, 41 cases died after withdrawal of treatment). The top five causes of death were abandonment of treatment due to financial reasons and the fear of adverse outcome (n=20), multiple organ dysfunction ( n =16 ) , and pneumothorax ( n =4 ) , diffuse intravascular coagulation (n=6), and shock (n=3). 41 cases survived were followed up for 9~54 months. The critical clinical conditions observed among these infants included cerebral palsy ( n = 5 ) , epilepsy ( n = 3 ) and developmental retardation(n=26).Conclusions There are many complications of severe HIE.The mortality of severe HIE is high, and the incidence of poor outcome of survivors is also high. Timely detection of risk factors is the key to the prevention of severe HIE. Long-term prognosis of severe HIE requires proper organization of neonatal follow up.

Objective Toinvestigatetheeffectofrehabilitationrobothandontheupperlimbmotor functioninpatientswithstrokeandhemiplegia.Methods Theeffectsoftraditionalrehabilitation therapy or robot rehabilitation treatment of 15 stroke patients with hemiplegia were analyzed retrospectively. There were 4 females and 11 males. According to the group order,they were divided into either a robot rehabilitation group (n=8 )or a general rehabilitation group (n =7 ). The patients of the general rehabilitation group received general rehabilitation training at the Department of Rehabilitation Medicine for 4 weeks,3 times a week,and 30 min for each time. At the same time,they also conducted family rehabilitation training,5 times a week,once for 1 h;the patients of the robot rehabilitation group received the general rehabilitation training for the same length of time as the general rehabilitation group,and they also performed robot rehabilitation training,5 times a week,once for 1 h. The patients of both groups were evaluated with the modified Ashworth scale (MAS )scores,Fugl-Meyer motor assessment of the upper extremity(FMA-UE),andWolfmotorfunctiontest(WMFT)beforeandaftertreatment.Results Compared with before training,the MAS score of the robot rehabilitation group was decreased significantly after four weeks of training. The time in WMFT was shortened significantly and the function score was improved obviously. The scores of upper extremity,hand score,and total FMA score in FMA-UE were improved significantly,and there were significant differences (all P <0. 05 ),while there were no significant differences in various indicators in the general rehabilitation group (all P>0. 05). Comparing the effects after four weeks of rehabilitation in both groups,the robot rehabilitation group was superior to the general rehabilitationgroupforthehandscoreinFMA-UE(allP<0.05).Conclusion Therehabilitation robot hand effectively lowered the upper limb spasticity. It has significant effect for recovery motor function of hand;however,it did not have a significant effect for motor rehabilitation on wrist.

Acid-Base Equilibrium ; Adolescent ; Child ; Drinking Water ; Humans ; Kidney ; Mineral Waters ; Minerals ; Schools ; Surveys and Questionnaires ; Water Supply

Humans ; Infant, Newborn ; Intestinal Perforation ; complications ; diagnosis ; Mycoses ; complications ; diagnosis

Objective To construct the engineering bacteria expressing the recombinant human Kunitz protease inhibitor domain of amyloid protein precursor variant (rhKD/APPvar)in Pichia pastoris,and to establish the methods suitable for large-scale fermentation and purification of rhKD/APPvar.Methods The rhKD/APPvar expression vector was constructed based on the rhKD/APPvar-pPICZαexpression vector. Two restriction enzyme loci (ApaⅠ and SacⅡ)were added to two flanks of KD/APP and human KD/APP activity center RAM was replaced by the active site of BPTI KAR.After the rhKD/APPvar-pPICZαexpression vector was transformed into Pichiapastoris,optimized expression and purification of rhKD/APPvar was performed.The rhKD/APPvar was purified with cation exchange chromatography and desalting.Results The results of digestion identification and DNA sequencing analysis demonstrated that the recombinant plasmid rhKD/APPvar-pPICZα was successfully constructed and transfected into pastoris X-33. The SDS-PAGE analysis results indicated that rhKD/APPvar expressed after the induction of methanol and the relative molecular weight was 6 700.After a series of experiments the optimal expression conditions of rhKD/APPvar were obtained as follows:the optimal pH was 6.0 and the optimal induction time point was about the 5 th day for the strain.After purified the purity of rhKD/APPvar was about 95%.Conclusion KD/APPvar-pPICZ is successfully constructed;after expression in Pichia pastoris and purification,the rhKD/APPvar protein is obtained.

OBJECTIVE: To investigate the effect of nasal instillation of vitamin D3 on patients with allergic rhinitis symptoms including nasal itching, sneezing, clear nasal discharge, and nasal congestion. METHOD: Thirty subjects with allergic rhinitis proved by skin prick test (SPT) and 30 subjects with deviated septum alone were recrui ted and administrated with 300 000 IU of vitamin D3 by nasal instillation weekly. Seven days after the intervention, the four major symptoms including nasal itching, sneezing, clear nasal discharge, and nasal congestion were evaluated by score. RESULT: After intranasal instillation of vitamin D3, the symptoms in allergic rhinitis group in cluding nasal itching, sneezing, nasal discharge and nasal congestion, and serum 25-hydroxyvitamin D level has statistical differences (P < 0.05). CONCLUSION Vitamin D3 could be well absorbed through nasal mucosa. It demonstrated to have significantly effect on serum 25-hydroxyvitamin D to improve the symptoms for patients with allergic rhinitis. Vitamin D3 may be a kind of adjuvant therapy for prevention and treatment of allergic rhinitis.
Administration, Intranasal ; Adult ; Cholecalciferol ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; drug therapy ; Young Adult