mRNA gene therapy has attracted much attention due to its advantages such as scalability, modification, no need to enter the nucleus and no integration of host genes. In gene therapy, safe and effective delivery of mRNA into cells is critical for the success of gene therapy. In this study, we designed and synthesized an amphiphilic cationic lipopeptide gene vector (dendritic arginine & disulfide bond-containing cationic lipopeptide, RLS) enriched with branched arginine. We achieved a 1.5-fold higher mRNA transfection efficiency in zebrafish compared to the commercial reagent Lipofectamine 2000, and confirmed its good biosafety by in vitro cytotoxicity and in vivo biosafety. First, we characterized the chemical composition of the cationic lipid peptides by nuclear magnetic resonance hydrogen spectroscopy (¹H NMR) and time-of-flight mass spectrometry (MS). The results of particle size and potential tested by dynamic light scattering particle size analysis showed that at a nitrogen/phosphorus (N/P) ratio of 20, the RLS/mRNA composite assemblies formed homogeneous nanoparticles with an average particle size of about 220 nm and a surface ζ potential of about +21 mV. In vitro gene transfection, the transfection experiments demonstrated that RLS exhibited 1.2-fold higher transfection efficiency in human embryonic kidney 293 cells (HEK293) and 3-fold higher transfection efficiency in rat mesenchymal stem cells (MSC) compared to Lipofectamine 2000. In addition, after microinjection of RLS into zebrafish embryos, we evaluated the survival, hatching, and teratogenicity rates, all of which confirmed its favorable in vivo safety profile. Thus, this amphiphilic cationic lipid peptide RLS, enriched with branched arginine, exhibits excellent mRNA delivery properties and safety. These findings highlight its potential as a promising gene therapy tool.

Objective To explore the effect of quercetin combined with iron death inhibitor Ferrostain-1 on oxalate-induced HK-2 cell injury.Methods HK-2 cells were randomly divided into control group(normal cultured cells),model group(0.5 mmol·L-1 calcium oxalate crystals),quercetin group(0.5 mmol·L-1 calcium oxalate crystals+100 μmol·L-1 quercetin),inhibitor group(0.5 mmol·L-1 calcium oxalate crystals+8 μmol·L-1 Ferrostain-1)and combination group(0.5 mmol·L-1calcium oxalate crystals+100 μmol·L-1quercetin+8 μmol·L-1 Ferrostain-1).Cell counting kit-8(CCK-8)assay was used to detect cell survival rate;Western blot was used to detect iron death related protein expression such as glutathione peroxidase 4(GPX4);flow cytometry and Tunel assay were used to detect cell apoptosis,and assay kit was used to detect cellular iron ions and antioxidant levels.Results The cell survival rates of control group,model group,quercetin group,inhibitor group and combination group were(100.00±2.55)％,(54.49±4.11)％,(64.26±6.30)％,(58.03±3.04)％and(79.37±4.29)％,respectively;GPX4 protein expression levels were 0.98±0.11,0.33±0.05,0.56±0.05,0.78±0.07 and 1.11±0.11,respectively;cell apoptosis rates were(4.15±0.28)％,(23.12±2.49)％,(17.28±1.07)％,(15.08±1.41)％and(8.95±0.75)％,respectively;Fe2+levels were(100.00±0.87)％,(162.55±14.70)％,(149.09±9.50)％,(144.95±11.12)％and(131.76±12.18)％,respectively;superoxide dismutase(SOD)levels were(58.67±3.46),(21.56±1.32),(33.60±2.03),(35.15±3.02)and(44.27±3.89)U·mL-1,respectively.The above indicators of the model group were compared with the control group,and the above indicators of the quercetin group,inhibitor group,and combination group were compared with the model group,the above indicators of the combination group were compared with the quercetin group,and inhibitor group,all they all showed statistical significance(all P＜0.05).Conclusion Iron death inhibitors can enhance the inhibitory effect of quercetin in vitro on oxalate-induced renal tubular epithelial cell injury.

Objective To investigate the effects of probiotic treatment combined with knowledge attitude and practice(KAP)model management on the recovery and quality of life of patients with severe acute pancreatitis.Methods Patients with severe acute pancreatitis from June 2020 to June 2023 in Quzhou People's Hospital were selected.The patients were divided into the control group,the probiotics group and the KAP combination group.All groups received symptomatic treatment.The control group was given routine nursing,the probiotic group was given daily oral administration of probiotic granules,and the KAP combination group was managed by KAP model on the basis of probiotic treatment,including targeted intervention in the dimensions of disease knowledge,behavior and belief.The APACHE Ⅱ scores,quality of life scale QOL-C30 scores,serum inflammatory factors C-reactive protein(CRP)and procalcitonin(PCT)levels were compared before and after treatment in three groups.Results A total of 108 patients were included,with 36 cases in each group.The CRP and PCT decreased significantly after treatment in three groups(P＜0.05).The reduction ranges of the probiotics group and the KAP group were significantly higher than that of the control group(P＜0.05).The self-management ability was significantly increased in all three groups after treatment compared to before treatment(P＜0.05),and the increase was significantly greater in the KAP combination treatment group than in the probiotic and control groups(P＜0.05).The APACHEⅡ scores decreased significantly and QOL-C30 scores increased significantly after treatment in the probiotics group and the KAP group(P＜0.05).Conclusion Probiotic treatment could significantly improve the inflammatory status,clinical symptoms and quality of life of patients with severe acute pancreatitis.The KAP model management could further enhance the effect of probiotic treatment,effectively improve patients'organism recovery ability and quality of life.

Objective:To detect the expression of angiogenic factor (AGGF1) with G-patch and FHA domain 1 in hepatocellular carcinoma (HCC), and to investigate its relationship with HCC angiogenesis, invasion and metastasis and its clinical significance.Methods:A total of 90 primary HCC patients were selected from the Huangshi Central Hospital and Hubei Province Cancer Hospital from January 2019 to September 2020. Clinicopathological data of all patients were collected. The expression of AGGF1 and CD34 in cancer tissues of 90 HCC patients was detected by immunohistochemistry, and microvascular density (MVD) was calculated. The patients were followed up for three years. The relationship between AGGF1 and serum angiopoietin-2 (Ang-2) content, MVD, clinicopathological parameters and postoperative prognosis was analyzed.Results:The positive rate, relative expression level and MVD value of AGGF1 in HCC tissues were significantly higher than those in adjacent tissues (all P<0.05). The relative expression of AGGF1 in cancer tissues was positively correlated with the MVD value ( r=0.716). Serum Ang-2 content and MVD in AGGF1-positive patients were significantly higher than those in AgGF1-negative patients (all P<0.01). AGGF1 expression in HCC tissues was significantly correlated with tumor differentiation, tumor size, (China Liver Cancer (CNLC) staging, lymph node metastasis, vascular invasion, and postoperative recurrence and metastasis (all P<0.01). Kaplan-Meier analysis showed that the 2- and 3-year survival rates of patients with high AGGF1 expression were significantly lower than those of patients with low AGGF1 expression (all P<0.01). Multivariate analysis showed that CNLC stage, lymph node metastasis, vascular invasion, AGGF1 expression, postoperative recurrence and metastasis were the influencing factors for postoperative survival of HCC patients (all P<0.05). Conclusions:The high expression of AGGF1 can promote the angiogenesis of HCC, and is significantly correlated with invasion and metastasis and poor prognosis, which can be a potential evaluation indicator of HCC clinical prognosis.

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 occupational exposure to blood-borne pathogens among medical workers in a tertiary general hospital in Hangzhou City from 2010 to 2022, so as to provide the evidence for improving occupational protective measures among medical workers. Methods: The registration and follow-up data of occupational exposure to blood-borne pathogens among medical workers from 2010 to 2022 were collected from the blood-borne occupational exposure monitoring system in a tertiary general hospital in Hangzhou City. The population distribution, occurrence, protection and disposal of occupational exposure to blood-borne pathogens were analyzed using a descriptive epidemiological method. Results: A total of 1 230 cases were reported with occupational exposure to blood-borne pathogens among medical workers in the study hospital from 2010 to 2022, with the highest incidence in 2021 (4.67%) and the lowest incidence in 2010 (0.99%). The incidence of occupational exposure to blood-borne pathogens appeared a tendency forwards a rise from 2010 to 2022 (P<0.05). Of all cases with occupational exposure to blood-borne pathogens, there were 934 women (75.93%), 656 nurses (53.33%), and 514 cases with working experiences of one year and shorter (41.79%). Hand was the predominant site of occupational exposure to blood-borne pathogens (92.03%) and ward was the predominant place of exposure (35.37%), while scalp needle was the predominant mode of exposure (32.68%), and removal of needle was the predominant procedure of exposure (32.36%). A total of 1 106 cases were tested for the blood-borne pathogens in the exposure sources, and 448 cases were tested positive for blood-borne pathogens, with a detection rate of 40.51%. Hepatitis B virus, treponema pallidum and human immunodeficiency virus were the three most common blood-borne pathogens, and there were 739 cases (60.08%) with personal protective equipment during exposure. Following the follow-up surveillance for more than 6 months post-exposure, no infections occurred. Conclusions Junior nurses and hand exposure were predominant among medical workers with occupational exposure to blood-borne pathogens in the study hospital from 2010 to 2022, and hepatitis B virus was the predominant blood-borne pathogen. No post-exposure infections occurred.

OBJECTIVE: To explore clinical efficacy of Ilizarov hemilateral bone longitudinal transport technique in treating hemilateral bone defects associated with chronic osteomyelitis of lower extremity long bones. METHODS: Clinical data of 13 patients with hemilateral bone defects caused by chronic osteomyelitis of lower extremity long bones and treated by Ilizarov hemilateral bone longitudinal transport technique were retrospective analyzed, including 10 males and 3 female, aged from 14 to 55 years old;4 patients occurred femoral and 9 patients occurred tibial;10 patients were diagnosed as traumatic osteomyelitis and 3 patients as hematogenous osteomyelitis. The anatomical classification of Cierny-Mader in 13 patients was type Ⅲ. Bone and wound healing, postopertaive complication, and bony and functional results were observed by Paley evaluation standard. RESULTS: After removing external fixator, all patients were followed up from 6 to 70 months. Transporting time ranged from 54 to 158 d. And the time in external fixation ranged from 6.8 to 19.5 months. External fixation index (EFI) ranged from 1.23 to 1.6 months/cm. According to Paley's evaluation criteria, bony results were excellent in 13 patients;functional results showed excellent in 12 patients and good in 1 patient. Two patients occurred poor union on the docking sites and healed with autogenous iliac bone graft. The callus at the extended area was poorly mineralized and improved significantly when treated with low-intensity pulsed ultrasound in one patient. All patients had good wound healing without recurrence of osteomyelitis and refracture. There was no vascular and nerve injury and axial deviation in all patients and they were satisfied with the appearance and function of lower limbs. The range of motion of knee and ankle joint before operation was 120 ° to 150 ° and 35 °to 80 ° respectively, and at the latest follow-up was 110 ° to 140 ° and 30 ° to 75 ° . CONCLUSION Ilizarov hemilateral bone longitudinal transport technique is effective in treating infective hemilateral bone defects of lower extremity long bones, which could not only simplify architecture of external fixation, but also reduce the number of fixation pins, shorten the time in external fixator and decrease the incidence of pin tract infection. However, this technique is highly demanding, and the growth of callus in extended region and healing of bone apposition should be noticed.
Male ; Humans ; Female ; Adolescent ; Young Adult ; Adult ; Middle Aged ; Retrospective Studies ; Lower Extremity/surgery* ; Tibia/surgery* ; Femur ; Ankle Joint

OBJECTIVE: This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province. METHODS: We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed. RESULTS: The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α ^3.7/αα (50.23%) and β ^IVS-II-654/β ^N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively. CONCLUSION Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans ; beta-Thalassemia/genetics* ; alpha-Thalassemia/genetics* ; Hemoglobinopathies/genetics* ; China/epidemiology* ; High-Throughput Nucleotide Sequencing

Objective:To investigate the effect of infliximab (IFX) on neurological function in mice after traumatic brain injury (TBI) and the role of nuclear factor-κB (NF-κB)/inducible nitric oxide lyase (iNOS) signaling in it.Methods:Seventy-two healthy adult male C57BL/6 mice were randomly divided into sham-operated group, TBI group, and TBI+IFX group ( n=24). The mouse TBI models were established by controlled cortical impact method. IFX (dissolved in normal saline at a concentration of 2.5 mg/mL and a dose of 10 μg/g) was administered intraperitoneally into the mice of TBI+IFX group 30 min after modeling once daily for 3 d; mice in the sham-operated group and TBI group were given the same amount of saline intraperitoneally at the same time points for 3 d. Neurological deficits (Garcia scores) were assessed one, 3 and 7 d after modeling; blood-brain barrier permeability was detected by Evans blue staining, and brain tissue water content was measured by dry and wet weight method; Nissl staining was used to detect the percentage of injured neurons in brain tissues; the percentage of apoptotic neurons was detected by Tunel staining; immunofluorescent double-labeling was used to detect the expressions of caspase-3 and neuronal nuclear antigen (NeuN) in neurons; immunohistochemical staining was used to detect the microglia marker ionized calcium binding adaptor molecule-1 (IBa-1) expression; ELISA was used to detect the expressions of inflammatory factors (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, interferon [IFN]-γ) and free radicals (oxygen free radicals [ROS], nitrogen free radicals [RNS]) in the brain tissues; and immunofluorescent staining and Western blotting were used to detect the expressions of nuclear factor (NF)-κB/inducible nitric oxide synthase (iNOS). Results:(1) One, 3 and 7 d after modeling, the Garcia scores showed significant differences among the three groups ( P<0.05); as compared with the TBI group, the TBI+IFX group had significantly increased Garcia scores 3 and 7 d after modeling ( P<0.05). (2) Three d after modeling, as compared with those in the TBI group, Evans blue leakage ([18.45±1.32] μg/g vs. [16.38±1.25] μg/g), brain water content ([81.56±0.96]% vs. [79.97±0.79]%), percentage of injured neurons ([79.50±5.85]% vs. [68.81±7.47]%), and percentage of apoptotic neurons ([41.93±7.49]% vs. [30.59±8.60]%) in mice of the TBI+IFX group were significantly deceased ( P<0.05). Three d after modeling, immunofluorescent double labeling showed that the relative caspase-3 expression in the TBI+IFX group (0.76±0.16) was significantly decreased as compared with the TBI group (1.11±0.23, P<0.05). Immunohistochemical staining and ELISA results showed that as compared with those in the TBI group, the Iba-1 staining scores, TNF-α, IL-1β, IL6 and IFN-γ levels, and ROS and RNS contents in TBI+IFX group were significantly decreased ( P<0.05). Immunofluorescent staining and Western blotting showed that as compared with the TBI group, the TBI+IFX group had significantly decreased expressions of NF-κB p65, iNOS and phosphorylated nuclear factor-κB inhibitor-α, and statistically inhibited nuclear translocation of NF-κB ( P<0.05). Conclusion:IFX can reduce inflammatory response and oxidative stress response, and play a neuroprotective role, which is related to its inhibition of downstream NF-κB/iNOS pathway activation.

OBJECTIVE: To compare treatment effect of monolateral and circular external fixator in infectious tibial nonunion and to explore external fixation structure with better osteogenic ability and fewer complications. METHODS: A retrospective analysis was performed on 150 patients with infectious tibial nonunion admitted from January 2010 to December 2014. Patients who met the inclusion and exclusion criteria were divided into monolateral fixator group and circular fixator group according to the type of external fixation. Demographic and perioperative data of the two groups were collected. New bone quality was assessed by pixels ratio, treatment effect was assessed by complications. Notes for treatment was explored by case analysis. RESULTS: A total of 64 patients with infectious mid-tibial nonunion were included, 26 in monolateral fixator group and 38 in circular fixator group. There was no difference in demographic data between two groups. The pixel ratio of new bone between two groups was from 0.91 to 0.97 with an average of (0.94±0.03), and from 0.93 to 0.97 with an average of(0.95±0.02), respectively, with no statistical significance (P>0.05). The external fixation index was from 34.1 to 50.6 with an average of (42.3±8.3) days /cm in monolateral fixator group and from 44.5 to 56.1 with an average of (45.8±10.3) days/cm in the circular fixator group, and the difference was not statistically significant (P>0.05). There were 7 cases (26.9%) of complications in monolateral fixator group and 5 cases (13.2%) in circular fixator group, the difference was not statistically significant (P>0.05), but 5 cases of foot ptosis in monolateral fixator group and none in circular fixator group. The time of weight bearing in monolateral fixator group was later than that in circular fixator group, and the difference was statistically significant (P<0.05). CONCLUSION Monolateral and circular fixators can achieve equivalent bone formation in the treatment of infected tibial nonunion. Circular fixator is recommended for patients with severe osteoporosis or concomitant medical diseases requiring early weight bearing. Hydroxyapatite coated screws are recommended if a monolateral external fixator is selected when the expected enlarged length is long and the fixation time is long and close monitoring of the ankle movement is required to avoid foot droop.
Humans ; Tibial Fractures/surgery* ; Retrospective Studies ; External Fixators ; Tibia/surgery* ; Treatment Outcome ; Hydroxyapatites