Objective To research the effect of molecular chimeric mice pre-T cells on proliferation ability of allogeneic mouse T cells.Methods The MHC-Ⅰ gene (H-2Kband H-2Db gene) were extracted and amplified by RT-PCR,the identified pre-T cells were transfected by the constructed eukaryotic expression vector of C57BL/6mouse MHC-I (pIRES-H-2Db and pIRES-H-2Kb),non-transfected group and sham pIRES-transfected control group were set.The molecular chimeric cells were transfused back to BALB/c mouse.After 7 days,T lymphocyte cells of each group were extracted,the ability of molecular chimeric cells inducing spleen T lymphocyte response to allogeneic T cells was observed through mixed lymphocyte culture (MLC).Results Sequencing of the plasmid we have constructed showed that insertion sequence contained C57BL/6 mice H-2Kb and H-2Db series which could be retrieved from GenBank.The result of flow cytometry analysis indicated that H-2Kb and H-2Db protein had an increased expression in pre-T cells,the difference with other two groups was statistically significant (P＜0.05).The result of MLC demonstrated that the stimulation index (SI) of T lymphocyte in co-injection transfected pIRES-H-2Kb and pIRES-H-2Db pre-T cells group (0.764±0.074) were significantly decreased compared with non-transfected group(0.983±0.081)and sham pIRES-transfected group(0.994±0.142) (P＜0.05).Conclusions The molecular chimeric pre-T cells infusion can reduce spleen T lymphocyte response to allogeneic T cells and it may induce immune tolerance in vivo.

Objective To establish the quality standard for Chanhou Zhuyu Capsules.Methods Leonurus Japonicus and Ligusticum chuanxiong in Chanhou Zhuyu Capsules were identified by TLC,and the content of stachydrine hydrochloride was determined by TLC Scanning.Results The quality identification by TLC was specific.Stachydrine hydrochloride had a good linearity in the range of 4.0～12.0 ?g,r=0.999 7,and the average recovery was 99.87 %,RSD=1.67 %.Conclusion The methods of identification and content determination are simple,reliable and reproducible.They can be used effectively for the quality control of Chanhou Zhuyu Capsules.

This study was aimed to observe influence on clinical symptoms of Qing-Fei Pei-Yuan Wei-Wan (QFPY-WW) for pulmonary infection HIV/AIDS cases with the syndrome of phlegm-heat obstructing the lung. A total of 141 pulmonary infection HIV/AIDS cases with the syndrome of phlegm-heat obstructing the lung were randomly divided into the treatment group (94 cases) and the control group (47 cases). The treatment group was given basic western medication combined with QFPYWW. The control group was given basic western medication. After 4 weeks treat-ment, observation was made on changes of main symptoms. The results showed that on the 28th day, compared with the control group, there was improvement on symptoms such as cough, breathing and chest tightness (P< 0.05). Com-parison on coughing up phlegm degree showed that the treatment group was significantly better than the control group (P< 0.05). On the 7th day treatment, the treatment group had better effect on lowering body temperature than the control group (P< 0.05). It was concluded that QFPYWW can improve symptoms such as cough, breathing, chest tightness and cough up phlegm among pulmonary infection HIV/AIDS patients.

PURPOSE: Sorafenib, a multikinase inhibitor, is the standard therapy for patients with advanced-stage hepatocellular carcinoma (HCC). However, resistance develops to the treatment, therefore, we tried to unravel the underlying mechanism in the resistance of HCC cells to sorafenib via the development of more effective therapeutic strategies. MATERIALS AND METHODS: Various liver cancer cell lines were treated with either sorafenib only or with sorafenib after infection of adenovirus expressing short hairpin RNA (shRNA) against transforming growth factor-β (TGF-β) and p38 activity was examined using western blotting. RESULTS: p38 MAP kinase activity was inhibited by low concentrations of sorafenib, which could potentially lead to sorafenib resistance in HCC cell lines. Subsequently, we used constitutive form of MKK3/6 (MKK3/6E) to confirm that massive cell death was induced by the activation of p38, and demonstrated the ability to activate p38 without any stimulation. In addition, sorafenib resistance was reduced by the activation of p38. Subsequently, we confirmed that TGF-β shRNA effectively recovered the phosphorylation of p38 inhibited by sorafenib, and increased the sensitivity of HCC cells to sorafenib, thereby inducing cell death and overcoming the resistance of HCC cells to sorafenib. CONCLUSION: Our study provides a new therapeutic strategy for HCC that overcomes the resistance of HCC to sorafenib by down-regulation of TGF-β.
Adenoviridae ; Blotting, Western ; Carcinoma, Hepatocellular* ; Cell Death ; Cell Line ; Down-Regulation* ; Humans ; Liver Neoplasms ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; RNA, Small Interfering

Objective:To investigate the differences in incidence of deep vein thrombosis (DVT) after closed fracture of lower extremity between patients with different blood types ABO.Methods:A retrospective study was conducted in the 1, 951 patients who had been admitted to Department of Orthopaedics Trauma, Honghui Hospital Affiliated to Xi'an Jiaotong University for lower extremity fractures from August 2014 to June 2018. They were 924 males and 1,027 females with a mean age of 63 (46, 78) years (range, from 16 to 102 years). Of them, 572 were type O, 564 type A, 609 type B and 206 type AB. Venous ultrasonography was performed on both lower extremities within 12 hours after admission. The incidences of DVT after fracture were compared between different blood types in all the patients, patients with proximal fracture of the knee, peri-knee fracture and distal fracture of the knee, and patients<60 years old and ≥60 years old.Results:The incidences of DVT were, respectively, 26.75% (153/572), 28.72% (162/564), 34.32% (209/609) and 29.61% (61/206) for patients with blood type O, type A, type B and type AB. The DVT incidence for type B was significantly higher than that for type O ( P< 0.008). The incidences of DVT were, respectively, 28.74% (98/341), 28.99% (100/345), 39.45% (144/365) and 30.97% (35/113) for blood type O, type A, type B and type AB in the patients with proximal fracture of the knee. The DVT incidence for blood type B was significantly higher than those for blood type O and blood type A ( P< 0.008). There were no significant differences in the DVT incidence between different blood types ABO in the patients with peri-knee fracture, distal fracture of the knee,<60 years old or ≥60 years old( P>0.05). The incidences of DVT were, respectively, 30.99% (97/313), 33.33% (108/324), 45.22% (156/345), 34.74% (33/95) for blood type O, type A, type B and type AB in the patients ≥60 years old. The DVT incidence for blood type B was significantly higher than those for blood type O and blood type A ( P< 0.008). Conclusions:The incidence of DVT varied with different blood types ABO after lower extremity fracture. The highest DVT incidence was found in patients with blood type B. The impact of blood type on the DVT incidence after lower extremity fracture was mainly observed in the patients with proximal fracture of the knee or an age of ≥ 60 years old.

Microbial transglutaminase, which could catalyze the cross-linking of many proteins or non-protein materials, has been widely used in food, pharmaceutical and textile industry. To enhance the yield of the enzyme and establish corresponding platform for molecular modification, the researchers of Japanese Ajinomoto began to construct the recombinant strain producing transglutaminase in the 1990s. So far, the enzyme has been successfully expressed in different expression systems. Some of the recombinant strains are more productive than wild strains. Recently, progress has been made in the molecular modification of microbial transglutaminase, and the activity, thermo-stability and specificity of the enzyme are improved. This review briefly summarized and analyzed the strategies involved in these studies, and noted its trends.
Bacterial Proteins ; biosynthesis ; genetics ; Catalysis ; Enzyme Stability ; Mutagenesis, Site-Directed ; Mutant Proteins ; genetics ; metabolism ; Protein Engineering ; methods ; Streptomyces ; enzymology ; Substrate Specificity ; Transglutaminases ; biosynthesis ; genetics

Objective:To evaluate the effect of age factors on the pharmacodynamics of intranasal dexmedetomidine for sedation in the pediatric patients undergoing transthoracic echocardiography(TTE).Methods:American Society of Anesthesiologists Physical Status classification Ⅰ-Ⅲ pediatric patients, aged 1-24 months, undergoing TTE from August 2019 to May 2022, were selected. This trial was performed in two parts. Part Ⅰ Pediatric patients were divided into 4 age groups: 1-6 month group, 7-12 month group, 13-18 month group and 19-24 month group. The initial dose of dexmedetomidine was 2.0 μg/kg in 0.1 μg/kg increment/decrement. The dose of dexmedetomidine was determined by using modified Dixon′s up-and-down method. The ED 50 and 95% confidence interval of intranasally administered dexmedetomidine for sedation were calculated by the Dexon-Massey method. Part Ⅱ One hundred patients were divided into 4 age groups ( n= 25 each): 1-6 month group, 7-12 month group, 13-18 month group and 19-24 month group. The 4 groups were further divided into 5 subgroups ( n=5 each) according to the dose of dexmedetomidine: 2.1 μg/kg subgroup, 2.2 μg/kg subgroup, 2.3 μg/kg subgroup, 2.4 μg/kg subgroup, and 2.5 μg/kg subgroup. Part Ⅰ and part Ⅱ trials were combined, and the ED 95 and 95% confidence interval of intranasally administered dexmedetomidine for sedation were calculated using the probit method. Results:A total of 220 pediatric patients were enrolled. There was no significant difference in ED 50 and ED 95 of dexmedetomidine intranasally administered for sedation among groups ( P>0.05). Conclusions:The pharmacodynamics of intranasal dexmedetomidine for sedation shows no significant difference in age in the pediatric patients aged 1-24 months undergoing TTE.

Objective:To investigate the clinical characteristics of patients with acute aortic dissection (AAD) through a retrospective and observational study, and to construct an early warning model of AAD that could be used in the emergency room.Methods:The data of 11 583 patients in the Emergency Chest Pain Center from January to December 2019 were retrospectively collected from the Chest Pain Database of Zhongshan Hospital Affiliated to Fudan University. Inclusion criteria: patients with chest pain who attended the Emergency Chest Pain Center between January and December 2019. Exclusion criteria were 1) younger than 18 years, 2) no chest/back pain, 3) patients with incomplete clinical information, and 4) patients with a previous definite diagnosis of aortic dissection who had or had not undergone surgery. The clinical data of 9668 patients with acute chest/back pain were finally collected, excluding 53 patients with previous definite diagnosis of AAD and/or without surgical aortic dissection. A total of 9 615 patients were enrolled as the modeling cohort for early diagnosis of AAD. The patients were divided into the AAD group and non-AAD group according to whether AAD was diagnosed. Risk factors were screened by univariate and multivariate logistic regression, the best fitting model was selected for inclusion in the study, and the early warning model was constructed and visualized based on the nomogram function in R software. The model performance was evaluated by accuracy, specificity, sensitivity, positive likelihood ratio and negative likelihood ratio. The model was validated by a validation cohort of 4808 patients who met the inclusion/exclusion criteria from January 2020 to June 2020 in the Emergency Chest Pain Center of the hospital. The effect of early diagnosis and early warning model was evaluated by calibration curve.Results:After multivariate analysis, the risk factors for AAD were male sex ( OR=0.241, P<0.001), cutting/tear-like pain ( OR=38.309, P<0.001), hypertension ( OR=1.943, P=0.007), high-risk medical history ( OR=12.773, P<0.001), high-risk signs ( OR=7.383, P=0.007), and the first D-dimer value ( OR=1.165, P<0.001), Protective factors include diabetes( OR=0.329, P=0.027) and coronary heart disease ( OR=0.121, P<0.001). The area under the ROC curve (AUC) of the early diagnosis and warning model constructed by combining the risk factors was 0.939(95 CI:0.909-0.969). Preliminary validation results showed that the AUC of the early diagnosis and warning model was 0.910(95 CI:0.870-0.949). Conclusions:Sex, cutting/tear-like pain, hypertension, high-risk medical history, high-risk signs, and first D-dimer value are independent risk factors for early diagnosis of AAD. The model constructed by these risk factors has a good effect on the early diagnosis and warning of AAD, which is helpful for the early clinical identification of AAD patients.

Objective:To compare the predictive value of the HEART, TIMI and GRACE scores for major adversecardiovascular events (MACEs) at 7 and 28 days in patients with actue non-ST-segment elevation myocardial infarction (NSTEMI).Methods:More than 12 000 patients with chest pain from the Emergency Department of Zhongshan Hospital Affiliated to Fudan University from October 2017 to October 2018 were studied, including 566 patients with cardiogenic chest pain, 105 patients with ST-segment elevation myocardial infarction (STEMI) excluded and 15 patients lost to follow-up. Finally, 109 patients with NSTEMI and 337 non-myocardial patients with cardiogenic chest pain were enrolled. NSTEMI patients were divided into subgroups according to whether MACEs occurred. LSD t-test, Mann-Whitney U test or χ2 test were used to analyze and compare the differences between the two subgroups about the baseline data, clinical data, HEART, TIMI and GRACE scores at the time of visit. Multivariate logistic regression analysis was used to explore the independent factors of MACEs at 7 and 28 days. And the predictive values of different scores for 7-day MACEs and 28-day MACEs were compared in NSTEMI patients through the receiver operating characteristic (ROC) curve. Results:Compared NSTEMI patients with non-myocardial patients with cardiogenic chest pain, we found a statistically significant differences in sex, past history of coronary heart disease,≥3 risk factors for atherosclerosis, electrocardiogram, high-sensitivity troponin T (hs-cTnT), creatinine value, past history of myocardial infarction, HEART score, TIMI score and GRACE score. In further subgroup analysis of NSTEMI patients who were divided according to whether MACEs occurred, we found previous history of stroke and increased hs-cTnT were statistically different in 7 days after the onset of the disease. The multivariate analysis showed that the previous history of stroke and increased hs-cTnT were independent factors for the occurrence of MACEs at 7 days after the onset of NSTEMI; The previous history of stroke and increased hs-cTnT, electrocardiogram ST segment depression and TIMI score were statistically different at 28 days after the onset of NSTEMI. The multivariate analysis showed that the previous history of stroke and TIMI score were independent factors for the occurrence of MACEs at 28 days after the onset of NSTEMI patients. ROC curve indicated that the predictive value of TIMI score (AUC=0.715, 95% CI: 0.482-0.948) was better than HEART (AUC=0.659, 95% CI: 0.414-0.904) and GRACE scores (AUC=0.587, 95% CI: 0.341-0.833)in predicting MACEs in NSTEMI patients. Conclusions:HEART score, TIMI score and GRACE score can be used to evaluate NSTEMI patients. There is an independent predictive value on TIMI score for the occurrence of 28-day MACEs in NSTEMI patients.

Objective:To study the association between preoperative hemoglobin amount and incidence of lower limb deep vein thrombosis (DVT) in patients with lower limb fracture.Methods:A retrospective study was performed of the 2, 482 patients with lower limb fracture who had been treated at Department of Orthopaedics Trauma, Honghui Hospital Affiliated to Xi'an Jiaotong University from July 2014 to August 2019. They were 1, 174 males and 1, 308 females with an age of (60.6±19.3) years. Recorded were the patients' age, gender, injury time, hemoglobin amount, D-dimer measurement, combined medical conditions, time and results of ultrasound vein examination on both lower extremities. According to the ultrasound results, the patients were divided into a thrombosis group and a thrombosis-free group. The 2 groups were compared in hemoglobin amount. Logistic regression was used to analyze the relationship between preoperative hemoglobin amount and incidence of lower limb DVT. The patients were divided into 5 groups according to the quintile of hemoglobin amount; the incidences of thrombosis were compared between the 5 groups.Results:The total incidence of DVT in this cohort was 29.53%(733/2, 482). The hemoglobin amount in the thrombosis group was (116.57±19.24) g/L, significantly lower than that in the thrombosis-free group (124.76±19.79) g/L ( P<0.05). The preoperative hemoglobin amount was a risk factor for incidence of DVT after a lower limb fracture ( OR=0.985, 95% CI: 0.980 to 0.990, P<0.001). As the quintile level of hemoglobin increased, the incidence of DVT showed a downward trend. In comparison of the group with the highest DVT incidence (40.58%) and the group with the lowest DVT incidence (17.27%), the risk increased by 2.386 times (95% CI: 1.718 to 3.315). Conclusions:The preoperative hemoglobin amount can affect the DVT incidence after a lower limb fracture, and a low hemoglobin amount may more likely lead to lower limb DVT.