[A cross-sectional study on the clinical phenotypes of rheumatoid arthritis].

OBJECTIVE: To explore the characteristics and clinical phenotypes of rheumatoid arthritis (RA) and provide the basis for further understanding, interventions and outcomes of this disease. METHODS: RA patients attended at Peking University People's Hospital from 2018 to 2021 were enrolled in the study. Data collection included demographic data, the sites and numbers of joints involved, extra-articular manifestations (EAM), comorbidities and laboratory variables. Statistical and bioinformatical analysis was performed to establish clinical subtypes by clustering analysis based on the type of joint involved, EAM involvement and other autoimmune diseases overlapped. The characteristics of each subtype were analyzed. RESULTS: A total of 411 patients with RA were enrolled. The mean age was (48.84±15.17) years, and 346 (84.2%) were females. The patients were classified into 4 subtypes: small joint subtype (74, 18.0%), total joint subtype (154, 37.5%), systemic subtype (100, 24.3%), and overlapping subtype (83, 20.2%). The small joint subtype had no medium or large joint involvement, and 35.1% had systemic involvement. The erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) levels and platelet count (PLT) were lower than those in other subtypes, and the rates of positive rheumatoid factors (RF-IgA and RF-IgG) were significantly higher in the small joint subtype. The total joint subtype had both large and small joint involvement but no systemic involvement. The rate of morning stiffness and positive antinuclear antibodies (ANA) in this subtype were lower than those in other subtypes. In the systemic subtype, interstitial lung disease and secondary Sjögren syndrome were the most common systemic involvements, with prominent levels of disease activity score 28-joint count (DAS28-ESR and DAS28-CRP). The overlapping subtype was commonly combined with Hashimoto's thyroiditis or primary Sjögren syndrome. Female in the overlapping subtype was more common than in other subtypes. This subtype was characterized by hyperglobulinemia, hypocomplementemia and high rate of positive ANA, especially spotting type. CONCLUSION: Based on the clinical features, RA patients could be classified into 4 subtypes: small joint subtype, total joint subtype, systemic subtype, and overlapping subtype. Each subtype had its own clinical characteristics. They help for further understanding and a more individualized treatment strategy of RA.

[Effects of exosomes from hepatocyte growth factor-modified human adipose mesenchymal stem cells on full-thickness skin defect in diabetic mice].

Objective: To investigate the effects and mechanism of exosomes from hepatocyte growth factor (HGF)-modified human adipose mesenchymal stem cells (ADSCs) on full-thickness skin defect wounds in diabetic mice. Methods: The experimental study method was adopted. Discarded adipose tissue of 3 healthy females (10-25 years old) who underwent abdominal surgery in the Department of Plastic Surgery of First Affiliated Hospital of Air Force Medical University from February to May 2021 was collected, and primary ADSCs were obtained by collagenase digestion method and cultured for 7 days. Cell morphology was observed by inverted phase contrast microscope. The ADSCs of third passage were transfected with HGF lentivirus and cultured for 5 days, and then the fluorescence of cells was observed by imaging system and the transfection rate was calculated. The exosomes of ADSCs of the third to sixth passages and the HGF transfected ADSCs of the third to sixth passages were extracted by density gradient centrifugation, respectively, and named, ADSC exosomes and HGF-ADSC exosomes. The microscopic morphology of exosomes was observed by transmission electron microscopy, and the positive expressions of CD9, CD63, and CD81 of exosomes were detected by flow cytometry, respectively. Twenty-four 6-week-old male Kunming mice were selected to make the diabetic models, and full-thickness skin defect wounds were made on the backs of mice. According to the random number table method, the mice were divided into phosphate buffer solution (PBS) group, HGF alone group, ADSC exosome alone group, and HGF-ADSC exosome group, with 6 mice in each group, and treated accordingly. On post injury day (PID) 3, 7, 10, and 14, the wounds were observed and the wound healing rate was calculated; the blood flow intensity of wound base was detected by Doppler flowmeter and the ratio of relative blood flow intensity on PID 10 was calculated. On PID 10, the number of Ki67 positive cells in wounds was detected by immunofluorescence method, and the number of new-vascularity of CD31 positive staining and tubular neovascularization in the wounds was detected by immunohistochemistry method; the protein expressions of protein endothelial nitric oxide synthase (eNOS), phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (Akt) and phosphorylated Akt (p-Akt) in wounds were detected by Western blotting, and the ratios of p-PI3K to PI3K and p-Akt to Akt were calculated. On PID 14, the defect length and collagen regeneration of wound skin tissue were detected by hematoxylin and eosin staining and Masson staining, respectively, and the collagen volume fraction (CVF) was calculated. The number of samples is 3 in all cases. Data were statistically analyzed with repeated measurement analysis of variance, one-way analysis of variance, and Tukey test. Results: After 7 days of culture, the primary ADSCs were spindle shaped and arranged in vortex shape after dense growth. After 5 days of culture, HGF transfected ADSCs of the third passage carried green fluorescence, and the transfection rate was 85%. The ADSC exosomes and HGF-ADSC exosomes were similar in microscopic morphology, showing vesicular structures with an average particle size of 103 nm and 98 nm respectively, and both were CD9, CD63, and CD81 positive. On PID 3, the wounds of mice in the 4 groups were all red and swollen, with a small amount of exudate. On PID 7, the wounds of HGF-ADSC exosome group were gradually reduced, while the wounds of the other three groups were not significantly reduced. On PID 10, the wounds in the 4 groups were all reduced and scabbed. On PID 14, the wounds in HGF-ADSC exosome group were basically healed, while the residual wounds were found in the other three groups. On PID 3, the healing rates of wounds in the four groups were similar (P>0.05); On PID 7 and 10, the wound healing rates in HGF-ADSC exosome group were significantly higher than those in PBS group, HGF alone group, and ADSC exosome alone group, respectively (with q values of 13.11, 13.11, 11.89, 12.85, 11.28, and 7.74, respectively, all P<0.01); on PID 14, the wound healing rate in HGF-ADSC exosome group was significantly higher than that in PBS group, HGF alone group, and ADSC exosome alone group (with q values of 15.50, 11.64, and 6.36, respectively, all P<0.01). On PID 3, there was no obvious blood supply in wound base of mice in the 4 groups. On PID 7, microvessels began to form in the wound base of HGF-ADSC exosome group, while the wound base of the other three groups was only congested at the wound edge. On PID 10, microvessel formation in wound base was observed in the other 3 groups except in PBS group, which had no obvious blood supply. On PID 14, the blood flow intensity of wound base in HGF-ADSC exosome group was stronger than that in the other 3 groups, and the distribution was uniform. On PID 10, the ratio of wound base relative blood flow intensity in HGF-ADSC exosome group was significantly higher than that in PBS group, HGF alone group, and ADSC exosome alone group (with q values of 23.73, 19.32, and 9.48, respectively, all P<0.01); The numbers of Ki67-positive cells and new-vascularity of wounds in HGF-ADSC exosome group were significantly higher than those in PBS group, HGF alone group, and ADSC exosome alone group, respectively (with q values of 19.58, 18.20, 11.04, 20.68, 13.79, and 8.12, respectively, P<0.01). On PID 10, the protein expression level of eNOS of wounds in HGF-ADSC exosome group was higher than that in PBS group, HGF alone group, and ADSC exosome alone group (with q values of 53.23, 42.54, and 26.54, respectively, all P<0.01); the ratio of p-PI3K to PI3K and the ratio of p-Akt to Akt of wounds in HGF-ADSC exosome group were significantly higher than those in PBS group, HGF alone group, and ADSC exosome alone group, respectively (with q values of 16.11, 11.78, 6.08, 65.54, 31.63, and 37.86, respectively, P<0.01). On PID 14, the length of skin tissue defect in the wounds of HGF-ADSC exosome group was shorter than that in PBS group, HGF alone group, and ADSC exosome alone group (with q values of 20.51, 18.50, and 11.99, respectively, all P<0.01); the CVF of wounds in HGF-ADSC exosome group was significantly higher than that in PBS group, HGF alone group and ADSC exosome alone group (with q values of 31.31, 28.52, and 12.35, respectively, all P<0.01). Conclusions: Human HGF-ADSC exosomes can significantly promote wound healing in diabetic mice by increasing neovascularization in wound tissue, and the mechanism may be related to the increased expression of eNOS in wounds by activating PI3K/Akt signaling pathway.

[Effects of exosomes from human adipose-derived mesenchymal stem cells on pulmonary vascular endothelial cells injury in septic mice and its mechanism].

Objective: To investigate the effects of exosomes from human adipose-derived mesenchymal stem cells (ADSCs) on pulmonary vascular endothelial cells (PMVECs) injury in septic mice and its mechanism. Methods: The experimental research method was adopted. The primary ADSCs were isolated and cultured from the discarded fresh adipose tissue of 3 patients (female, 10-25 years old), who were admitted to the First Affiliated Hospital of Air Force Medical University undergoing abdominal surgery, and the cell morphology was observed by inverted phase contrast microscope on the 5th day. The expressions of CD29, CD34, CD44, CD45, CD73, and CD90 of ADSCs in the third passage were detected by flow cytometry. The third to the fifth passage of ADSCs were collected, and their exosomes from the cell supernatant were obtained by differential ultracentrifugation, and the shape, particle size, and the protein expressions of CD9, CD63, tumor susceptibility gene 101 (TSG101), and ß-actin of exosomes were detected, respectively, by transmission electron microscopy, nano-particle tracking analysis and Western blotting. Twenty-four adult male BALB/c mice were adopted and were divided into normal control group, caecal ligation perforation (CLP) alone group, and CLP+ADSC-exosome group with each group of 8 according to random number table (the same grouping method below) and were treated accordingly. At 24 h after operation, tumor necrosis factor (TNF-α) and interleukin 1ß (IL-1ß) levels of mice serum were detected by enzyme-linked immunosorbent assay, and lung tissue morphology of mice was detected by hematoxylin-eosin and myeloperoxidase staining, and the expression of 8-hydroxy-deoxyguanosine (8-OHdG) of mouse lung cells was detected by immunofluorescence method. Primary PMVECs were obtained from 1-month-old C57 mice regardless gender by tissue block method. The expression of CD31 of PMVECs was detected by immunofluorescence and flow cytometry. The third passage of PMVECs was co-cultured with ADSCs derived exosomes for 12 h, and the phagocytosis of exosomes by PMVECs was detected by PKH26 kit. The third passage of PMVECs were adopted and were divided into blank control group, macrophage supernatant alone group, and macrophage supernatant+ADSC-exosome group, with 3 wells in each group, which were treated accordingly. After 24 h, the content of reactive oxygen species in cells was detected by flow cytometry, the expression of 8-OHdG in cells was detected by immunofluorescence, and Transwell assay was used to determine the permeability of cell monolayer. The number of samples in above were all 3. Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results: The primary ADSCs were isolated and cultured to day 5, growing densely in a spindle shape with a typical swirl-like. The percentages of CD29, CD44, CD73 and CD90 positive cells of ADSCs in the third passage were all >90%, and the percentages of CD34 and CD45 positive cells were <5%. Exosomes derived from ADSCs of the third to fifth passages showed a typical double-cavity disc-like structure with an average particle size of 103 nm, and the protein expressions of CD9, CD63 and TSG101 of exosomes were positive, while the protein expression of ß-actin of exosomes was negative. At 24 h after operation, compared with those in normal control group, both the levels of TNF-α and IL-1ß of mice serum in CLP alone group were significantly increased (with t values of 28.76 and 29.69, respectively, P<0.01); compared with those in CLP alone group, both the content of TNF-α and IL-1ß of mice serum in CLP+ADSC-exosome group was significantly decreased (with t values of 9.90 and 4.76, respectively, P<0.05 or P<0.01). At 24 h after surgery, the pulmonary tissue structure of mice in normal control group was clear and complete without inflammatory cell infiltration; compared with those in normal control group, the pulmonary tissue edema and inflammatory cell infiltration of mice in CLP alone group were more obvious; compared with those in CLP alone group, the pulmonary tissue edema and inflammatory cell infiltration of mice in CLP+ADSC-exosome group were significantly reduced. At 24 h after operation, endothelial cells in lung tissues of mice in 3 groups showed positive expression of CD31; compared with that in normal control group, the fluorescence intensity of 8-OHdG positive cells of the lung tissues of mice in CLP alone group was significantly increased, and compared with that in CLP alone group, the fluorescence intensity of 8-OHdG positive cells in the lung tissues of mice in CLP+ADSC-exosome group was significantly decreased. The PMVECs in the 3rd passage showed CD31 positive expression by immunofluorescence, and the result of flow cytometry showed that CD31 positive cells accounted for 99.5%. At 12 h after co-culture, ADSC-derived exosomes were successfully phagocytose by PMVECs and entered its cytoplasm. At 12 h after culture of the third passage of PMVECs, compared with that in blank control group, the fluorescence intensity of reactive oxygen species of PMVECs in macrophage supernatant alone group was significantly increased (t=15.73, P<0.01); compared with that in macrophage supernatant alone group, the fluorescence intensity of reactive oxygen species of PMVECs in macrophage supernatant+ADSC-exosome group was significantly decreased (t=4.72, P<0.01). At 12 h after culture of the third passage of PMVECs, and the 8-OHdG positive fluorescence intensity of PMVECs in macrophage supernatant alone group was significantly increased; and compared with that in blank control group, the 8-OHdG positive fluorescence intensity of PMVECs in macrophage+ADSC-exosome supernatant group was between blank control group and macrophage supernatant alone group. At 12 h after culture of the third passage PMVECs, compared with that in blank control group, the permeability of PMVECs monolayer in macrophage supernatant alone group was significantly increased (t=6.34, P<0.01); compared with that in macrophage supernatant alone group, the permeability of PMVECs monolayer cells in macrophage supernatant+ADSC-exosome group was significantly decreased (t=2.93, P<0.05). Conclusions: Exosomes derived from ADSCs can ameliorate oxidative damage in mouse lung tissue, decrease the level of reactive oxygen species, 8-OHdG expression, and permeability of PMVECs induced by macrophage supernatant.

[Influencing factors of thrombosis besides antiphospholipid antibodies in patients with antiphospholipid syndrome].

Objective: To identify the influencing factors of thrombosis besides antiphospholipid antibodies in patients with antiphospholipid syndrome (APS). Methods: The 169 patients diagnosed with APS were enrolled according to the current APS classification criteria from January 2003 to August 2017 in Peking University People's Hospital. There were 23 males and 146 females with a mean age of (41±15) years. Antiphospholipid antibodies, including anticardiolipin (aCL), anti-ß2glycoprotein-1 (ß2GP1) antibodies and antibodies to the phosphatidylserine-prothrombin complex (aPS/PT), were determined by enzyme-linked immunosorbent assay (ELISA) methods. Lupus anticoagulant (LA) was identified using the STA Compact coagulation testing system. The differences of clinical and laboratory characteristics between patients with and without thrombosis were analyzed (100 cases and 69 cases, respectively). The influencing factors for thrombosis in patients with APS were determined using binary logistic regression. Results: Compared with patients without thrombosis, patients with thrombosis were older and had a longer disease duration ((45±17) years vs (35±9) years and M(Q1, Q3) 12.0(3.8, 84.0) months vs 48.0(12.0, 108.0) months, both P<0.05). The percentage of male, primary APS, smoking, low blood platelet count, hypertension, and diabetes in patients with thrombosis were significantly higher than those in patients without thrombosis (all P<0.05). Similarly, the rates of antinuclear antibodies positive, aCL positive, aPS/PT-IgM positive, and aPS/PT-IgG positive in patients with thrombosis were significantly higher than those in patients without thrombosis (all P<0.05). The levels of D-dimer in patients with thrombosis were significantly higher than that in patients without thrombosis (P<0.05). There was significant difference in global anti-phospholipid syndrome score (GAPSS) between patients with and without thrombosis (P<0.05). The GAPSS score was also significantly higher in patients with arterial thrombosis than that in patients with venous thrombosis (P<0.05). Smoking and D-dimer levels were independent influencing factors for thrombosis in patients with APS (smoking: OR=11.222, 95%CI:1.119-112.544, P=0.040, D-dimer levels: OR=1.002, 95%CI: 1.000-1.003, P=0.037). Conclusions: Thrombotic APS patients are older and have a longer suffering duration, a higher ratio of male, primary APS, smoking, hypertension, lower blood platelet count, diabetes, higher GAPSS scale, and higher D-dimer levels. Smoking and D-Dimer levels may be independent risk factors for thrombosis in patients with APS.

Accuracy Analysis of Distinguishing the Cooperation Degree during Image Completion Test by Eye Movement Parameters.

ABSTRACT: Objective To analyze the differences in accuracy of different eye movement parameters in distinguishing the cooperation and non-cooperation during image completion test of patients with mental disorders caused by craniocerebral trauma. Methods One hundred and forty cases of patients with mental disorders caused by craniocerebral trauma who took psychiatric impairment assessments were collected. The 21 pictures from "image completion" of Wechsler intelligence test were used as stimulating pictures, then divided into cooperation group and non-cooperation group according to binomial forced-choice digit memory test and expert opinions. The eye movement parameters of research subjects during completion of images were obtained by the SMI eye-tracker. The accuracy of eye movement parameters in distinguishing the cooperation or non-cooperation of patients with mental disorders caused by craniocerebral trauma in psychiatric impairment assessments were evaluated by the ROC curve. Results During the process of the image completion test, the area under curve （AUC） value of frequency of blink, frequency of fixation, pupil size, frequency of saccade, latency of saccade, average acceleration of saccade, the average and peak longitudinal velocity of saccade was above 0.5. When it comed to a specific stimulating picture, the AUC value of frequency of blink in looking at a specific stimulating picture could be above 0.8, and the AUC value of X axis diameter of pupil size could be above 0.7. Conclusion The accuracy of eye movement parameters in distinguishing the cooperation or disguise of patients with mental disorders caused by craniocerebral trauma is related with the stimulating picture. The accuracy of frequency of blink in distinguishing cooperation and non-cooperation is better than that of other eye movement parameters.

Working Memory of Patients with Mild Cognitive Impairment due to Brain Trauma Based on fNIRS.

ABSTRACT: Objective To discuss the activation characteristics of the prefrontal cortex of people with mild cognitive impairment （MCI） due to brain trauma during working memory tasks. Methods The psychological experiment design software E-prime was used and N-back paradigm was adopted as working memory task. Functional near-infrared spectroscopy （fNIRS） was used to detect changes in cortical oxygenated hemoglobin concentrations of 22 channels within the prefrontal lobe of 24 people with MCI due to brain trauma （study group） and 27 healthy volunteers （control group） with matching gender and age. Behavioral data, such as the number of keystroke errors and reaction time, were recorded simultaneously. Independent samples t test and non-parametric test were used to compare the mean value of oxygenated hemoglobin concentration change, the number of key errors and the mean value of reaction time of the two groups in each task. Results （1） The differences in the number of errors and reaction time between the two groups in 1-back and 2-back tasks had statistical significance （P<0.05）.The main effects of task load and group were both significant （task F=14.11, P=0.001 1; group F=10.39, P=0.001 5）. （2） During the 1-back task, the differences in oxygenated hemoglobin concentration changes of the 22 channels between the two groups had no statistical significance （P>0.05）. During the 2-back task, the differences in oxygenated hemoglobin concentration changes of the two groups in channel 2, 3, 7, 9, 10, 11, 14, 15, 18, 19, 21 and 22 had statistical significance （P<0.05）. （3） In the 1-back task, the left frontal pole and dorsolateral prefrontal area in both groups were activated. In the 2-back task, the activation areas of the control group were the left frontal pole area and the left dorsolateral prefrontal area, while that of the study group almost covered most of the left and right frontal pole areas, which were scattered and the right area was activated, too. Conclusion Patients with MCI due to brain trauma have obvious working memory impairment, and during the 2-back working memory task, the activation of the prefrontal lobe decreased, but the activation range was wider.

Application of Mismatch Negative in Evaluation of Severity of Mental Disorders due to Traumatic Brain Injury.

ABSTRACT: Objective To explore the applied value of mismatch negative （MMN） in evaluation of severity of mental disorders due to traumatic brain injury. Methods Thirty-five patients（case group） that conform to the diagnostic criteria of organic （traumatic brain injury） mental disorder in ICD-10 Classification of Mental and Behavioural Disorders criteria were selected. Twenty-four healthy subjects （normal control group） that matched the case group in terms of gender, age composition ratio and educational level were selected. All subjects were evaluated by Activity of Daily Living Scale （ADL） and Social Disability Screening Schedule （SDSS） and then examined by Event-Related Potential （ERP）. A statistical analysis of the data was made by SPSS 22.0 software. Results The 32 patients and 24 normal control subjects completed the study. The scores of ADL and SDSS were significantly higher in the case group than in the normal control group （P<0.05）. The latency of Fz, FCz, Cz and Pz in the case group was significantly longer than that in the normal control group （P<0.05）. In the case group, the latency of Fz, FCz, Cz and Pz was positively correlated with the scores of ADL and SDSS （P<0.05）. The equation can be well fitted with the scores of ADL and SDSS. The latency and amplitude of Fz, FCz, Cz and Pz were used as concomitant variables and whether or not the subjects had mental disorders due to traumatic brain injury as dependent variables. Conclusion The latency of MMN can be used as an indicator in potential evaluation of the severity of mental disorders due to traumatic brain injury, which means that the longer the latency of MMN is, the more severe mental disorders due to traumatic brain injury may be. The combined application of ADL, SDSS and MMN can be an objective indicator in preliminary judgment of mental disorders due to traumatic brain injury.

[Application of Activities of Daily Living Scale in Mild Psychiatric Impairment Assessment].

OBJECTIVES: To explore the application of activities of daily living （ADL） scale in mild psychiatric impairment assessment under the guideline of Classification of Human Body Disability Caused by Injury. METHODS: A total of 124 subjects with organic mental disorders and mild psychiatric impairments （levels 7 to 10）, and 106 healthy controls were included in. All participants were assessed by the ADL scale, physical self-maintenance scale （PSMS） and instrumental activities of daily living （IADL） scale. The difference between the scores of control group and study group, and the relationship of impairment level and the scores were compared, and the threshold value was determined according to the ROC curve. RESULTS: The total scores of ADL, IADL and PSMS were significantly different between the control group and the study group （P<0.05）. The scores of ADL, IADL, PSMS were significantly different among the impairment levels （P<0.05）, which showed a relativity with impairment level. The scores of ADL corresponding to levels 10, 9, 8 and 7 were 14-17, 18-23, 24-29 and 30-34, respectively, which showed a good correlation between the conclusion according to the scale and the expert's opinion （κ= 0.914, P<0.05）. CONCLUSIONS: The score of ADL was significantly related to mild psychiatric impairment, and the higher ADL score represents the more severe disability, which can be used as a reference index for preliminarily judging the level of mild psychiatric impairment.

[Effects of microRNA-34a on regulating silent information regulator 1 and influence of the factor on myocardial damage of rats with severe burns at early stage].

Objective: To explore the effects of microRNA-34a on regulating silent information regulator 1 (SIRT1) and influence of SIRT1 on myocardial damage of rats with severe burns at early stage. Methods: (1) Twenty-four Sprague-Dawley (SD) rats were divided into sham injury (SI) group, simple burns (SB) group and SIRT1 agonist (SA) group according to the random number table (the same grouping method below), with 8 rats in each group. Rats in groups SB and SA were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burns) on the back, and rats in group SI were sham injuried on the back. Immediately after injury, rats in groups SI and SB were intraperitoneally injected with normal saline of 50 mL/kg, and rats in group SA were intraperitoneally injected with normal saline of 50 mL/kg and 1 mg/mL resveratrol of 50 mg/kg. At 6 h post injury, abdominal aortic blood was collected to make serum and myocardial tissue of rats was collected. (2) Myocardial cells of twelve neonatal SD rats were collected and divided into microRNA-34a mimic control (MMC) group, microRNA-34a mimic (MM) group, microRNA-34a inhibitor control (MIC) group, and microRNA-34a inhibitor (MI) group, which were respectively transfected with gene sequences of mimic control, mimic, inhibitor control, and inhibitor of microRNA-34a. The microRNA-34a expression level and protein expression level of SIRT1 in myocardial cells were respectively detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Another batch of myocardial cells were divided into microRNA-34a inhibitor control+ burn serum (MCB) group, microRNA-34a inhibitor+ burn serum (MB) group, and microRNA-34a inhibitor+ burn serum + EX527 (MBE) group. Myocardial cells in group MCB were transfected with gene sequence of inhibitor control, and myocardial cells in the later groups were transfected with gene sequence of inhibitor of microRNA-34a. After transfection of 48 h, myocardial cells in group MBE were cultured in Dulbecco's modified Eagle's medium (DMEM) solution for 6 hours, with serum in group SB of volume fraction of 10% and final amount-of-substance concentration of 1 mol/L, and myocardial cells in the other 2 groups were cultured in DMEM solution with serum from rats of group SB of volume fraction of 10%. The protein expression levels of myocardial cells of SIRT1, cleaved-caspase-3, and Bax were detected by Western blotting. (3) Myocardial tissue from (1) was collected to detect expression levels of microRNA-34a and mRNA of SIRT1 in groups SI and SB by real-time fluorescence quantitative RT-PCR. Morphology of myocardial tissue of rats in groups SI, SB, and SA was observed with biological image navigator. The mRNA expression levels of interleukin 1ß (IL-1ß) and tumor necrosis factor (TNF-α) of rats in groups SI, SB, and SA were detected by real-time fluorescence quantitative RT-PCR. The expression levels of cleaved-caspase-3, and Bax of myocardial tissue of rats in groups SI, SB, and SA were detected by Western blotting. Data were processed with one-way analysis of variance and least-significant difference test. Results: (1) After transfection of 48 h, the expression level of microRNA-34a of myocardial cells in group MM was 4.67±0.92, significantly higher than 1.03±0.04 in group MMC (P<0.01); the protein expression level of SIRT1 of myocardial cells in group MM was 0.35±0.06, significantly lower than 1.12±0.11 in group MMC (P<0.01). After transfection of 48 h, the expression level of microRNA-34a of myocardial cells in group MI was 0.26±0.07, significantly lower than 1.33±0.07 in group MIC (P<0.01); the protein expression level of SIRT1 of myocardial cells in group MIC was 1.12±0.16, significantly lower than 1.74±0.34 in group MI (P<0.01). At 6 h after culture, compared with those in group MCB, the SIRT1 protein expression level of myocardial cells in group MB was significantly increased (P<0.05), while cleaved-caspase-3 and Bax protein expression levels of myocardial cells in group MB were significantly decreased (P<0.05). Compared with those in group MB, the SIRT1 protein expression level of myocardial cells in group MBE was with no significantly statistical difference (P>0.05), and cleaved-caspase-3 and Bax protein expression levels were significantly increased (P<0.05). (2) At 6 h post injury, compared with that in group SI, the microRNA-34a expression level of myocardial tissue in group SB was significantly increased (P<0.01), and the mRNA expression level of SIRT1 of myocardial tissue in group SB was significantly decreased (P<0.01). At 6 h post injury, myocardial cells in group SI arranged neatly with normal nucleus and no inflammatory cells infiltration; myocardial cells in group SB arranged disorderly, with no abnormal nucleus, and obvious inflammatory cells infiltration; myocardial cells in group SA arranged neatly, with normal nucleus and little inflammatory cells infiltration. At 6 h post injury, compared with those in group SB, the mRNA expression levels of IL-1ß and TNF-α, and the protein expression levels of cleaved-caspase-3 and Bax of myocardial tissue in groups SI and SA were significantly decreased (P<0.01). Conclusions: The microRNA-34a expression level of myocardial tissue of rats with severe burns at early stage increases, which decreases the expression level of SIRT1, and increases the expression levels of IL-1ß, TNF-α, cleaved-caspase-3 and Bax, leading to obvious myocardial damage. Activation of SIRT1 can alleviate myocardial damage of rats with severe burns at early stage through decreasing expression levels of IL-1ß, TNF-α, cleaved-caspase-3, and Bax.

[Eye Movement Characteristics of Cooperation Degree during Image Completion Test in Psychiatric Impairment Assessment].

OBJECTIVES: To explore the difference of eye movement characteristics between uncooperative and cooperative subjects with mental disorder after cerebral trauma. METHODS: Thirty-nine subjects which needed psychiatric impairment assessment were selected. According to the binomial forced-choice digit memory test （BFDMT）, all subjects were divided into cooperative and uncooperative groups. The subjects were asked to take the image completion test from Wechsler adult intelligence scale. Meanwhile, the data of eye movement track, fixation, saccade, pupil and blink were recorded by the track system of eye movement. RESULTS: There were significantly differences （P<0.05） in the data of saccade between cooperative （10 cases） and uncooperative groups （29 cases）. The frequency, time, amplitude, acceleration of saccadic in uncooperative group were significantly higher than cooperation group. The saccade latencies of cooperation group increased more than uncooperative group. There was a significant difference （P<0.05） in total discrete distance, average distance and total time of fixation between two groups, while the average duration time, number and frequency of fixation had no significantly difference （P>0.05） between two groups. And the blink frequency of cooperation group was higher than uncooperative group. CONCLUSIONS: Eye movement can be an objective index for the primary judgment of cooperation level.

[Applied Value of Electroencephalogram in Assessment of Mild Psychiatry Impairment].

OBJECTIVES: To explore the applied value of electroencephalogram （EEG） in assessment of psychiatric impairment among patients with mental disorders due to traumatic brain injury. METHODS: According to the ICD-10, a total of 271 subjects were enrolled and assessed with the criterion of mental disorders due to traumatic brain injury. Activity of Daily Living Scale （ADL）, Functional Activities Questionnaire （FAQ） and Social Disability Screening Schedule （SDSS） were used to evaluate the severity of patients. All the participants were tested by Wechsler Adult Intelligence Scale （WAIS） and examined by EEG. RESULTS: Totally 215 patients accomplished the study. The results of Glasgow Coma Scale （GCS）, the severity of craniocerebral injury and the scores of FAQ, SDSS and ADL showed significant difference among the patients with different severity of EEG （P<0.05）. The grades of psychiatric impairment showed significant difference among the patients with different abnormal EEG （P<0.05）. CONCLUSIONS: EEG can reflect the severity of craniocerebral injury, assist evaluate the social function and activity of daily living of patients with mental disorders due to traumatic brain injury, and distinguish the mild psychiatric impairment grades, which suggest that EEG has a certain reference value in the assessment of psychiatric impairment.

[Effects of activating silent information regulator 1 on early kidney damage in rats with severe burn].

Objective: To investigate the effects of activating silent information regulator 1 (SIRT1) on the early kidney damage in rats with severe burn. Methods: Thirty healthy male SD rats were divided into sham injury group (SI), pure burn group (PB), and SIRT1 activator group (SA) according to the random number table, with 10 rats in each group. Rats in groups PB and SA were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burn) on the back. Immediately after injury, rats in group PB were intraperitoneally injected with normal saline in the dosage of 50 mL/kg, and those in group SA with 1 mg/mL (final mass concentration) resveratrol in the dosage of 50 mL/kg. Rats in group SI were sham injured and intraperitoneally injected with normal saline in the dosage of 50 mL/kg immediately after injury. Kidney tissue and abdominal aorta blood of rats in the three groups were collected at 24 hours after injury. The morphology of kidney tissue was observed after HE staining. The serum content of creatinine and urea nitrogen was determined with enzyme-linked immunosorbent assay. Protein expressions of SIRT1, Bax, and Bcl-2 in kidney tissue were determined with Western blotting. mRNA expressions of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß), and IL-10 in kidney tissue were determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with one-way analysis of variance and LSD-t test. Results: (1) In rats of group SI, structures of kidney tubules and glomeruli were intact. In rats of group PB, structures of kidney tubules were not clear with casts in them, and glomeruli showed pyknosis. In rats of group SA, structures of kidney tubules were relatively intact, and the pyknosis of glomeruli were slighter as compared with that of group PB with fewer glomeruli showing pyknosis. (2) The serum content of creatinine and urea nitrogen in rats of group PB was (67±14) µmol/L and (22.0±4.4) mmol/L, respectively, which was significantly higher than that of group SI [(28±7) µmol/L and (5.5±1.2) mmol/L respectively, with t values respectively 6.07 and 11.53, P values below 0.01]. The serum content of creatinine and urea nitrogen in rats of group SA was (39±9) µmol/L and (14.1±1.7) mmol/L, respectively, significantly lower than that of group PB (with t values respectively 4.09 and 4.17, P values below 0.01). (3) Compared with those of group SI, protein expressions of SIRT1 and Bcl-2 in kidney tissue of rats in group PB were significantly decreased (with t values respectively 16.32 and 19.58, P values below 0.01), while the protein expression of Bax was significantly increased (t=5.98, P<0.01). Compared with those of group PB, protein expressions of SIRT1 and Bcl-2 in kidney tissue of rats in group SA were significantly increased (with t values respectively 6.94 and 5.37, P values below 0.01), while the protein expression of Bax was significantly decreased (t=3.44, P<0.01). (4) mRNA expressions of TNF-α, IL-1ß, and IL-10 in kidney tissue of rats in group PB were 17.0±4.0, 2.27±0.59, and 2.5±0.9, respectively, significantly higher than those of group SI (1.0, 1.00, and 1.0, respectively, with t values from 3.27 to 8.93, P<0.05 or P<0.01). mRNA expressions of TNF-α and IL-1ß in kidney tissue of rats in group SA were 6.8±1.2 and 1.18±0.26, respectively, significantly lower than those of group PB (with t values respectively 4.59 and 4.32, P values below 0.01). mRNA expression of IL-10 in kidney tissue of rats in group SA was 5.0±1.0, significantly higher than that of group PB (t=5.51, P<0.01). Conclusions: Activating SIRT1 on early stage of severe burn in rats can decrease levels of creatinine and urea nitrogen, thus improving the kidney function. It can down-regulate the protein expression of Bax and up-regulate the protein expression of Bcl-2, thus reducing the apoptosis in kidney tissue. Meanwhile, it can inhibit expressions of TNF-α and IL-1ß and promote the expression of IL-10, thus alleviating the inflammatory response in kidney.