Objective To investigate the survival profile of the intradermally injected mouse autologous skin fibroblasts and the changes of the collagen fibers by using green fluorescent protein labeling and two-photon fluorescence microscopy. Methods The cultured cells were transfected by EGFP lentivirus, and then the cells were injected into the corresponding mouse skin. Biopsy was taken from the animals after 1 and 2 months. The specimens made serial frozen sections, the survival profile of the injected cells and the changes of the collagen fibers were observed by two-photon fluorescence microscopy. The collagenic area and dermal thickness were measured with image analysis software, and statistical analysis was also carried out. Results Two-photon fluorescence microscopy showed clear images of the injected cells and collagen fibers. Both the area of collagen fibers and the dermal thickness were significantly increased in injected cells after 2 months (P<0.05), however, there were no difference between injected cells and control after 1 mouth (P>0.05). Conclusions Autologous cultured fibroblasts could survive in a long time after transplantating into the skin, and collagen could be newly produced, the depth of dermis increases, which provides a possibility to treat mini-defects of the tissue.

Objective To investigate the effect of the Ras-related tumor suppressor gene aplasia Ras homolog member Ⅰ (ARHI) on angiogenesis in hepatocellular carcinoma (HCC).Methods We generated stable cell lines overexpressing ARHI in Hep3B cells,which lack endogenous ARHI.Cell proliferation was assessed by the MTT assay.The effects of ARHI overexpression on tumor growth and angiogenesis were assessed.Because of the key role of mammalian target of rapamycin (mTOR)signaling in HCC progression,we also tested whether ARHI overexpression affected the mTOR pathway.Results Ectopic expression of ARHI significantly diminished cell proliferation in Hep3B cells (P＜0.01).ARHI overexpression significantly retarded Hep3B xenograft growth by 76.4 ％ in vivo,and caused a marked reduction in tumor angiogenesis assessed by CD31-stained microvessel count.Western blot analysis of the xenografts showed that ARHI overexpression substantially reduced the phosphorylation of two mTOR substrates,S6K1 and 4E-BP1,indicative of an inactivation of the mTOR pathway.Accompanying with the mTOR inactivation,the angiogenic factors,hypoxia-inducible factor 1 alpha and vascular endothelial growth factor,were significantly downregulated.Conclusion These data highlighted an important role for ARHI in controlling HCC growth and angiogenesis,therefore offering a possible therapeutic strategy against this malignancy.

Objective:To analyze the changes of cerebral perfusion level and its relationship with cognitive impairment in patients with first anterior circulation acute ischemic post-stroke cognitive impairment.Methods:From March 2018 to March 2020, 70 patients with acute ischemic stroke in the first anterior circulation who were treated in the Department of Neurology of Kailuan General Hospital affiliated to North China University of Technology and met the inclusion and exclusion criteria were retrospectively analyzed.The mini-mental state examination was used to divide 36 cases of post-stroke cognitive impairment group and 34 cases of post-stroke cognitive impairment group.All the selected patients were examined by computed tomography(CT) and diffusion-weighted MR imaging(MRI-DWI) to determine the diagnosis and distribution of lesions; detection of cerebral artery stenosis by MRA; apply 3D pseudo-continuous arterial spin labeling(3D-pCASL) perfusion imaging, set the post label delay 1.5 s and 2.5 s to detect cerebral perfusion level.Results:(1)There was no statistical significance between the two groups in the comparison of basic clinical data.(all P>0.05). (2) The proportion of patients with middle cerebral artery, anterior cerebral artery and internal carotid artery ≥1 or ≥2 moderate and severe stenosis / occlusion on the focal side in the post-stroke cognitive impairment group (91.67%(33/36), 33.33%(12/36)) was higher than that in the post-stroke non-cognitive impairment group (23.53%(8/34), 8.82%(3/34)); the proportion of patients with moderate and severe stenosis / occlusion of MCA and ICA on the focal side in the post-stroke cognitive impairment group (69.44%(25/36), 44.44%(16/36)) was higher than that in the post-stroke cognitive impairment group (14.71%(5/34), 11.76%(4/34)), and the difference was statistically significant(χ 2 values were 33.455, 6.239, 21.394, 9.150, all P<0.05). (3) The proportion of patients with ≥ 2 cerebral infarction lesions in the post-stroke cognitive impairment group (61.1%(22/36))was higher than that in the post-stroke cognitive impairment group (38.03%(27/71))than that in the non-stroke group (20.6%(7/34), 19.05%(8/42)), and the difference was statistically significant(χ 2=11.833, 4.447, all P<0.05). PLD 2.5 s, the CBF value of frontal lobe infarction in post-stroke cognitive impairment group((31.516±8.333) mL/(100 g·min)) was lower than that in post-stroke non-cognitive impairment group((45.442±8.281) mL/(100 g·min)), the difference was statistically significant( t=3.835, P<0.05). Correlation analysis showed that the CBF value of PLD 2.5 s frontal infarction lesion was positively correlated with MMSE score( r=0.738, P<0.05). (4) The proportion of patients with ≥ 1 or ≥ 2 hypoperfusion areas (frontal lobe, temporal lobe, parietal lobe) on the focal side of PLD 1.5 s and 2.5 s after stroke (88.89%(32/36), 88.89%(32/36), 77.78%(28/36), 66.67%(24/36)) were higher than those without cognitive impairment after stroke (67.65%(23/34), 8.82%(3/34), 29.41%(10/34), 0), the difference was statistically significant(χ 2 values were 4.686, 44.837, 16.483, 34.493, all P<0.05). At PLD 1.5 s, CBF values of frontal lobe and parietal lobe in cerebral hypoperfusion area ((20.260±5.266) mL/(100 g·min), (17.664±3.947) mL/(100 g·min)) in patients with cognitive impairment after stroke were lower than those in patients without cognitive impairment ((33.442±10.563) mL/(100 g·min), (28.071±6.913) mL/(100 g·min)), the difference was statistically significant( t values were 3.392, 6.225, all P<0.05), at PLD 2.5 s, the CBF value after compensatory perfusion of frontal lobe, parietal lobe and temporal lobe in the post-stroke cognitive impairment group ((37.732±8.355) mL/(100 g·min), (32.942±6.459) mL/(100 g·min), (39.282±7.443) mL/(100 g·min)) was lower than that in the non cognitive impairment Group ((57.189±9.965) mL/(100 g·min), (52.415±7.017) mL/(100 g·min), (49.258±8.912) mL/(100 g·min)), the difference was statistically significant( t values were 5.443, 10.227, 2.950, all P<0.05). Correlation analysis found that the CBF value of the frontal lobe and parietal lobe of the PLD 1.5 s lesion area and the CBF value of the PLD 2.5 s hypoperfusion brain area after the perfusion of the frontal lobe, parietal lobe, and temporal lobe were positively correlated with the MMSE score( r values were 0.693, 0.675, 0.823, 0.799, 0.545, all P<0.05). Conclusion:Patients with first anterior circulation acute ischemic post-stroke cognitive impairment often have extensive hypoperfusion in the peripheral cerebral region, the occurrence of cognitive impairment after the first anterior circulation acute ischemic post-stroke cognitive impairment is related to the decrease of the perfusion level of the infarct lesion and the brain area around the lesion.

Objective To observe the effect of puerarin on the ICAM-1 gene expression in streptozotocin-Induced diabetic nephropathy rats. Methods Diabetic nephropathy rats were induced by intraperitoneal injection of STZ.Rats were allocated into normal control group,model group,and puerarin group.The puerarin treatment lasted for 16 weeks.During and after the treatment,the general health conditions,blood glucose levels,blood urea nitrogen,serum creatinine,urinary albumin excretion rate of 24-hours,and clearance rate of creatinine of the rats were observed.The ICAM-1mRNA expressions were also determined by in situ hybridization analysis in the kidney tissue of the rats.Results Diabetes mellitus and renal function lesion occurred in the model group and puerarin group.Puerarin could improve the general health of the rats,decrease blood urea nitrogen,serum creatinine,and urinary albumin excretion rate of 24-hours;and increase the clearance rate of creatinine.The ICAM-1mRNA expressions in the kidney were significantly inhibited by puerarin treatment.Conclusion Puerarin has a function of renal protection.

Objective To analyze the clinical characteristics of Duchenne/ Becker muscular dystrophy (DMD/BMD)with the initial presentation of transaminase elevation,in order to improve the clinician's understanding of this disease,and reduce misdiagnosis and missed diagnosis. To investigate the relationship between the elevation of transami-nase and the early stage of DMD/ BMD,and to provide the strategy for early diagnosis. Methods Twenty - four pa-tients admitted to the hospital with elevated serum transaminase as the initial presentation from January 2012 to Decem-ber 2014,who were diagnosed as DMD/ BMD by genetic testing or muscle biopsy,were enrolled. Their clinical data and laboratory examinations were retrospectively analyzed,including clinical features,diagnostic steps,serum muscle en-zymes,genetic analysis,electromyography and muscle pathological changes. Results The 24 patients were all male without family history of DMD/ BMD prior to birth. The average visiting age was (3. 4 ± 1. 2)years (ranging from 0. 8 to 6. 1 years),and 87. 5％ (21 / 24 cases)of cases were preschool children aged 2 - 6 years. Hypertransaminasemia was found in 21 cases (87. 5％)during the kindergarten physical examination,1 case during pre - operative investigation and 2 cases during respiratory infection. Due to its insidious onset,the time interval between incidental finding of elevated transaminase and definitive diagnosis was between 0. 6 and 20. 4 months. Among them,16 cases (66. 7％)had obvious pseudohypertrophy of calf muscles,and 18 cases (81. 8％)showed different degrees of movement disorder,such as unable to jump,easy to fall,and difficulty in climbing stairs. In addition,18. 2％ cases (4 / 22 cases)had a delay in language development. The serum alanine aminotransferase and aspartate aminotransferase levels were 120. 3 - 761. 7 U/ L and 83. 3 - 675. 5 U/ L,respectively. Serum creatine kinase (CK)was found to be markedly elevated (ranging from 3940 to 27510 U/ L)in all patients. Electromyography showed myogenic damage in 13 / 23 cases (56. 5％). DMD gene deletions were found in 18 cases (75. 0％),and duplications in 4 cases (16. 7％). The muscle biopsy performed in 2 cases (8. 3％)multiplex ligation - dependent probe amplification (MLPA)- negative cases showed evidence of dystrophic features on routine histology. Immunohistochemistry showed absent dystrophin staining in all 2 MLPA - nega-tive cases. Conclusion The clinical manifestation of DMD/ BMD is not typical in the early stage,so it is easy to be neglected or misdiagnosed. Elevated ALT and AST are important clues in the early diagnosis of DMD/ BMD. Children with elevated transaminase,in the absence of other signs and symptoms of hepatic injury,may have occult muscular di-sease,most frequently DMD/ BMD. A thorough physical examination and history taking as well as the measurement of serum CK are helpful in the differential diagnosis. Genetic testing or muscle biopsy should be done early for correct diagnosis of DMD/ BMD.

Objective:To construct a chronic obstructive pulmonary disease (COPD) assessment test (CAT) score prediction model based on a deep network fused with air data, and to explore its significance.Methods:From February 2015 to December 2017, the outdoor environmental monitoring air data near the residential area of the patients with COPD from the Respiratory Outpatient Clinics of Peking University Third Hospital, Peking University People′s Hospital and Beijing Jishuitan Hospital were collected and the daily air pollution exposure of patients was calculated. The daily CAT scores were recorded continuously. The CAT score of the patients in the next week was predicted by fusing the time series algorithm and neural network to establish a model, and the prediction accuracy of the model was compared with that of the long short-term memory model (LSTM), the LSTM-attention model and the autoregressive integrated moving average model (ARIMA).Results:A total of 47 patients with COPD were enrolled and followed up for an average of 381.60 days. The LSTM-convolutional neural networks (CNN)-autoregression (AR) model was constructed by using the collected air data and CAT score, and the root mean square error of the model was 0.85, and the mean absolute error was 0.71. Compared with LSTM, LSTM-attention and ARIMA, the average prediction accuracy was improved by 21.69%.Conclusion:Based on the air data in the environment of COPD patients, the fusion deep network model can predict the CAT score of COPD patients more accurately.