Objective To study the In vitro expansion of human CD8+ CD28- suppressor T cells and their immunological regulatory effect.Methods Human CD8 + CD28- suppressor T cells were expanded in vitro driven by the combination of cytokines and allogeneic antigen presenting cells (APCs).Flow cytometry was used to assess the development of CD28- subpopulation.Expanded CD8+ CD28- T cells were isolated by immunomagnetic microspheres and then added as third part modulators into mixed lymphocyte culture to assess their immunological regulatory characteristics.Results The combination of cytokines included IL-2,IL-7 and IL-15 and allogeneic APCs could increase the portion of CD8 + CD28- T cell subtype,and expansion fold of CD8+ CD28- T cell subtype was significantly increased as compared with others (P＜0.05).Expanded CD8+ CD28- T cells could suppress the proliferation of CD4+ T cells stimulated by allogeneic APCs.Moreover,this suppression had antigen specificity.Conclusion Human CD8 + CD28- suppressor T cells can be in vitro expanded in large amounts driven by the combination of cytokines and allogeneic APCs.Expanded CD8 + CD28- T cells in this study have antigen specific regulatory characteristics.

Small Animal Radiation Research Platform (SARRP) is a highly efficient platform specifically designed for research in the fields of radiation oncology and radiobiology.This platform possesses precise radiometers with gantries capable of isocentric,coplanar,and non-coplanar contormal irradiation.Based on the three-dimensional images acquired by the cone-beam computed tomography,the platform can perform real time treatment planning and irradiation,which assures timeliness for precise irradiation and research on small animals.The wide application of SARRP not only accelerates the bench to bedside translation,but also facilitates the development of radiation oncology and radiobiology.

Currently,the toxicity study of traditional Chinese medicine is faced with the following problems. Firstly,the evaluation in vitro cannot fully reflect the true state of the body. Secondly,the traditional method is not sensitive enough to the early toxicity. Lastly,the toxicity evaluation indexes cannot determine whether the compatibility of traditional Chinese medicine produces toxicity or increases toxicity systematically. The paper proposed a synthesized early evaluation research method for target organ toxicity induced by traditional Chinese medicine:screening,validation,optimization and application. This method mainly inoolves early target organ toxicity biomarkers in screening,optimi?zation,validation,biological significance explanation,and application to the traditional Chinese medicine incompatibility based on the metabolic dynamic fingerprint spectrum in order to obtain biomarkers of target organ toxicity that are sensitive and precede conventional biochemical indices for early evaluation . We attempted to analyze the pattern of chang of the biomarkers for animals acted by″18 incompatible medicaments″compatibility combination. We found that Radix Aconiti Lateralis Preparata with cardiotoxicity were compatible with Rhizoma Pinelliae,and that Trichosanthes kirilowii Maxim,Fritillaria,Ampelopsis Radix and Bletilla striata without non-cardiotoxicity produced and increased cardiotoxicity systematically.

Objective To explore the effectiveness of our self-designed 3D printing relay template in the treatment of malunion of irregular intra-articular ankle fractures.Methods From October 2013 to October 2015,our self-designed 3D printing relay template was used in 14 patients with severe malunion of intra-articular ankle fracture to assist their intra-articular osteotomy.They were 10 males and 4 females,with an average age of 40.5 years (from 34 to 59 years).Thin slice CT scan was preformed preoperatively for all the ankles involved.The computer fictitious tibial and fibular models for location and osteotomy navigation templates were acquired from the data of CT scan after processing,design and reconstruction by software.After the navigation templates which fit the size of solid ankle were printed by a 3D printer,they were sterilized and reserved.The guide plates were used in turn during operation.The irregular malunited bone blocks in the posterior malleolus were completely resected along the osteotomy template.After fracture reduction,the bone blocks were stabilized with internal fixation.Fracture union time was recorded.Scores of visual analogue scale (VAS),American Orthopaedic Foot and Ankle Society (AOFAS) and the short form-36 (SF-36),ankle ROM and complications were also recorded at the final follow-ups.Results The 14 patients obtained an average follow-up of 28.6 months (from 14 to 35 months).Bony union was achieved after an average of 3.7months (from 3 to 6 months).At the final follow-ups,on average,VAS score (2.2 ± O.8),AOFAS ankle and hindfoot score (81.1 ± 6.8),SF-36 score (82.5 ± 5.2),dorsal extension (27.5 ° ± 2.5°),and plantar flexion (24.2° ±6.3°) were significantly improved compared with the preoperative values (6.3 ± 1.8,43.1 ± 12.0,37.6 ± 9.9,14.1° ± 3.8° and 14.4° ± 3.9°,respectively) (P ＜ 0.05).No implant failure,soft tissue complications or post-traumatic arthritis occurred during the follow-up.Conclusion Since our self-designed 3D printing relay templates for osteotomy can accurately position the fracture lines of the distal tibia via limited incision and invasion,they help a lot in precise osteotomy and anatomical reduction of malunited intra-articular ankle fracture,preventing incidence of traumatic arthritis.

OBJECTIVETo investigate the feasibility of using the concentration of 1,2-DCE in blood as a biological monitoring indicators and build the determination method of 1,2-DCE in blood.

METHODSDose-response relationship of the exposure of 1,2-DCE and the level of 1,2-DCE in rat blood were investigated using the Pearson's correlation analysis. The concentration of 1,2-DCE in blood was determined using Headspace Sampler-Gas Chromatography-Mass Spectrometer (HS-GC-MS). 3.0 ml blood sample diluting with 2.0 ml 1,2-DCE standard serial solution was placed in 15 headspace bottles respectively and heated at 80 ℃ for 20 min.The vapor upon the headspace bottle was separated by capillary column and the concentration of 1,2-DCE was determined by massspectrum in SIM mode to draw a standard work curve. The within-run precision and the between-run precision were calculated by the relative standard deviation (RSD) of the concentration of 1,2-DCE in blood which was determined 6 times in a day and 6 times within 3 days respectively. The recovery rate was calculated by P=(C2 -C0)/C1 × 100%.

RESULTSWhen the treatment groups were exposed at dosage of 1,472, 2,550, 3,093, 3,976, and 4 418 mg/m(3), the average concentration of 1,2-DCE in rat blood was 24.1,231.6,344.3,395.1,538.5 μg/L. There was a positive correlation between the concentration of 1,2-DCE in rat blood and the exposed level of 1,2-DCE.The equation of dose-response relationship was y=0.162x -195.8,r=0.982 2,P=0.003 and the precision of exposure experiments was 7.04% -13.15%. 1,2-DCE contents within 0.259 -2 587 μg/L showed a good linear relationship and the regression equation was y=47 901x -357 446, r= 0.999 8. When the blood containing 0.259 μg/L 1,2-DCE was determined for six times, the average peak/peak signal-to-noise ratio was 56.55. The limit of detection (LOD) was 0.014 μg/L and the limit of quantification (LOQ) was 0.046 μg/L. The within-run precision was 1.23% -2.76% and the between-run precision was 2.21% -4.64%. The average recovery rate was 93.3% - 98.6%.

CONCLUSIONThe concentration of 1,2-DCE in blood could be used as a biological monitoring indicator. The method of the concentration of 1,2-DCE in blood determining by HS-GC-MS was characterized by high sensitivity, wide linear range, small interference, high precision and easy operation.

Animals ; Environmental Exposure ; analysis ; Environmental Monitoring ; Ethylene Dichlorides ; blood ; Gas Chromatography-Mass Spectrometry ; Limit of Detection ; Rats

To summarize the biological effects of four natures of traditional Chinese medicine (TCM) in the recent 40 years, including cold, hot, warm and cool, which have the effect on central nervous system, endocrine system on the activity of sympathetic nerves, basal metabolic rate, the function of organs and tissues and secretion of cytokine out of body. And to review the new concepts and new hypotheses in recent 10 years which proposed on the four natures of traditional Chinese medicine. Based on the above summary we pointed out that the previous studies on biological effects have shortage in the following aspects: The studies had little connection with the TCM theory; The research highly concentrated on the biological effects of the cold and heat natures of drug; There is almost no research on the biological effects of the neutral nature; The research on the biological effects of the natures of traditional Chinese medicine had no combination with effects of drugs. And pointed out that studies on four natures of traditional Chinese medicine will be a multi-level, multi-disciplinary, multi-factor, multi-targe research, connecting Chinese medicine theory. The research will be a combination of the macro research and the micro research, the qualitative research and the quantitative research and the experimental research and the clinical research.
Medicine, Chinese Traditional ; methods

The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen.Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nueleoeapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonai serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses.The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.

OBJECTIVETo assess the association of HLA-DPA1 and -DPB1 polymorphisms with Posner-Schlossman syndrome (PSS) in southern Chinese Han population.

METHODSA total of 100 randomly selected PSS patients of southern Chinese Han origin were served as the experimental group, while 128 unrelated healthy blood donors of the same origin were served as the control group. All samples were subjected to sequencing-based typing (SBT) for exon 2 of HLA-DPA1 and -DPB1 loci in both directions. HLA genotype was assigned using an Assign 3.5 HLA SBT software. The allele frequencies and haplotype frequencies of HLA-DPA1 and -DPB1 of the two groups were compared. x² test, P value and odds ratio (OR) value were calculated.

RESULTSSix HLA-DPA1 alleles in the experimental group and 4 HLA-DPA1 alleles in the healthy control group were identified. The allelic frequency for HLA-DPA1*02:01 in the experimental group was significantly lower than the control group (4.50% vs. 12.109%; x²=8.124, P=0.004). Sixteen HLA-DPB1 alleles were identified in both the experimental and control groups. The allelic frequencies for HLA-DPB1*14:01 and - DPB1*17:01 in the experimental group were significantly lower than those of the control group ( DPB1*14:01: 1.00% vs. 4.688%, x²=5.130, P=0.024; DPB1*17:01: 0% vs. 2.344%, x²=3.897, P=0.048). The DPA1-DPB1 haplotypes for the experimental and control groups were 23 and 25, respectively. The haplotype frequencies for both DPA1*02:01- DPB1*14:01 and DPA1*02:01- DPB1*17:01 were significantly lower than those of the control group.

CONCLUSIONDPA1*02:01- DPB1*14:01 and DPA1*02:01- DPB1*17:01 haplotypes may provide considerable protection effect against PSS in the southern Chinese Han population.

Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; Case-Control Studies ; China ; ethnology ; Female ; Glaucoma ; ethnology ; genetics ; HLA-DP alpha-Chains ; genetics ; HLA-DP beta-Chains ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic

Objective To evaluate the value of the ratio of optic nerve sheath diameter to eyeball transverse diameter (ONSD/ETD) in assessing intracranial pressure using ultrasound in the patients with brain injury.Methods Forty-six patients of both sexes with brain injury in the surgical intensive care unit,were selected.Craniotomy was performed within 24 h after admission to hospital,and the intracranial pressure probe was placed in the lateral cerebral ventricle of the enrolled patients who were aged 18-80 yr,with Glasgow Coma Scale score 3-15.ONSD and ETD were measured within 3 days after operation,three times a day,and the intracranial pressure was simultaneously recorded.The correlation between ONSD/ETD ratio and intracranial pressure were analyzed using the Pearson correlation analysis.The receiver operating characteristic curve was used to evaluate the accuracy of ONSD/ETD ratio in assessing the intracranial hypertension (intracranial pressure＞20 mmHg).Results The ONSD/ETD ratio was positively correlated with intracranial pressure (r =0.720,P＜0.01).The area under the receiver operating characteristic curve was 0.900 (95％ confidence interval 0.854-0.946),and the threshold was 0.248 (sensitivity 89.4％,specificity 77.5％).Conclusion The ONSD/ETD ratio produces higher accuracy in assessing intracranial pressure in the patients with brain injury.

Objective:To explore the related factors influencing prognosis of patients with grade Ⅲ cholangitis with myocardial injury.Methods:91 patients with grade III cholangitis complicated with myocardial injury treated in the emergency department of Beijing Friendship Hospital Affiliated to Capital Medical University from June 2015 to December 2020 were collected retrospectively. They were divided into endoscopic retrograde cholangiopancreatography (ERCP) group ( n=75) and non-ERCP group ( n=16) according to whether ERCP was performed. According to 28-day survival patients were divided into survival group ( n=56) and death group ( n=35). The level of serum cardiac troponin T (cTnT), cardiac troponin I (cTnI), creatine kinase isoenzyme (CK-MB), N terminal pro-brain natriuretic peptide (NT-proBNP), white blood cell (WBC), platelet (PLT), serum creatinine (Scr), alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin (TBIL), International Standardized Ratio (INR), Glasgow Coma Score (GCS) at different time points were detected and the risk factors influencing the prognosis of patients were analyzed. Results:The cTnT ( P=0.018), NT-proBNP ( P<0.001), PLT ( P=0.016), GCS score ( P=0.07) on day 3 and cTnI ( P=0.027), cTnT ( P=0.002), CK-MB ( P=0.046), NT-proBNP ( P<0.001), PLT ( P=0.041), GCS score ( P<0.001) on day 7 in the ERCP group were significantly different with the non-ERCP group respectively. The survival rate within 28 days of the ERCP group was significantly different from that in the non-ERCP group ( P<0.001). The cTnT ( P=0.006) on day 1, the cTnT ( P=0.021), NT-proBNP ( P=0.02), WBC ( P=0.037), GCS score ( P<0.001) on day 3, and the cTnI ( P=0.029), cTnT ( P=0.008), CK-MB ( P<0.001), PLT ( P=0.008), NT-proBNP ( P=0.004), GCS ( P<0.001) on day 7 in survival group were significantly different from the death group. Logistic regression showed that the mean value of myocardial injury markers and ERCP process were significantly correlated with the 28 days survival rate. Conclusions:In patients with Grade Ⅲ cholangitis-related myocardial injury, the levels of myocardial injury markers, NT-proBNP, platelet (PLT) and Glasgow Coma Score (GCS) are related to mortality. ERCP for patients can significantly improve 28 days survival and prognosis.