BACKGROUND: At present, there is little related report about producing a whole-kidney acellular matrix (ACM) scaffold in rats using perfusion. The cellular biocompatibility of the ACM is poorly understood. OBJECTIVE: To produce a whole-kidney ACM scaffold in rats by perfusion, to evaluate the cytocompatibility of ACM with the L929 cells in vitro, and to assess the possibility of ACM as the cytoskeleton and tissue-engineered urinary organ construction. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory of Zhujiang Hospital, Southern Medical University from February to May 2009. MATERIALS: Kidneys were obtained from 12-week-old Whista rats, while ureter, renal veins and renal artery were reserved. Intravenous catheters were inserted through renal arteries to establish channels for perfusion. Whole-kidney retrograde perfusion was performed with successively heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 9.81 kPa to prepare whole-kindney acellular matrix scaffolds. METHODS: ① Samples were randomly divided into blank group (without any cells), negative control group (culture media), experimental group (rat kidney ACM leaching liquor), and positive control group (culture media containing 0.64% phenol). L929 cells in the logarithmic phase were seeded in 96-well plates at the density of 4×10~3/well, with 5 wells in each group. At 24, 72, and 120 hours after incubation, cells were stained with MTT method to detect absorbance at 490 nm and calculate relative growth rate. ② Control group (culture medium), experimental group (rat kidney ACM leaching liquor), and positive control group (culture media containing 0.64% phenol) were set up to detect cell apoptosis at 48 hours after culture using flow cytometry. MAIN OUTCOME MEASURES: Microstructure of the scaffold, cytotoxicity and cell apoptotic rate. RESULTS: After SDS and TritonX-100 union processing, reticulate structures made of basilar membrane and collagen were shown under scanning electron microscope rather than normal structures of cells. At every time intervals (24, 72, and 120 hours), there was no significant difference in the absorbance between experimental group and negative control group (P > 0.05). The grade of the cytotoxicity of the ACM was .0-1. There was no significant difference in cell apoptotic rate between experimental group and negative control group (P > 0.05). CONCLUSION: The whole-kidney acellular matrix scaffolds in rat by perfusion have good biocompatibility.

Objective To analyze the electroencephalograph (EEG) features of 43 patients with clinically possible or probable Creutzfeldt-Jakob disease (CJD) and its correlation with megnetic resonance imaging (MRI) imaging and clinical manifestations.Methods All patients diagnosed with suspected CJD who were hospitalized in Xuanwu Hospital from January 2013 to December 2015 were collected.The clinical data, EEG and MRI imaging features were analyzed retrospectively.Based on the periodic sharp wave complexes (PSWC) appearance in EEG results, the patients were divided into typically changed group (TCG), atypically changed group (ACG) and unchanged group (UCG).Age, disease duration, clinical manifestations and MRI features among three groups were analyzed and the correlations between patient′s EEG features and age, disease duration, clinical manifestations or MRI features were explored using spearman method.Results Among the 43 patients with possible or probable CJD disease, 26 were male and 17 were female with an average disease duration of 4 months.The age of onset ranged from 31 to 80 with an average of (58.0±9.8) years old, and 86.0% of patients were 51 years old or above.Clinical characteristics of CJD patients according to occurrence rate were as follows: 35 cases (81.4%) with cognitive impairment, 29 cases (67.4%) with mental and behavior disorder, 28 cases (65.1%) with pyramidal tract damage, 24 cases (55.8%) with cerebellar symptoms, 23 cases (53.5%) with extrapyramidal symptoms,17 cases (39.5%) with myoclonic, 13 cases (30.2%) with dyssomnia, 13 cases (30.2%) with visual disorder and 2 cases (4.7%) with akinetic mutism.Regarding EEG features, 39.53% (17/43) of patients showed typical periodic sharp wave complexes (PSWC) (TCG group), 51.2% (22/43) had irregular rhythm and different forms of slow wave (ACG group) and only 9.3% (4/43) had no EEG change (UCG group).The occurrence rate of ribbon sign in MRI was 82.4% (14/17) in TCG group, 77.3% (17/22) in ACG group and none in UCG group.The rates were significantly higher in TCG and ACG group than that in UCG group (both P<0.05).Spearman correlation analysis revealed that EEG features was correlated with disease duration (r=0.351, P=0.021) and visual impairment (r=-0.377, P=0.013) for all CJD patients.There was no correlation between EEG and MRI or other clinical manifestations such as myoclonic, age and so on (all P>0.05).Conclusions EEG showed typical changes associated with disease duration in different stages of disease.EEG and MRI are two different means to evaluate different aspects of patients with CJD disease, and combination of two means could achieve better evaluation results.

OBJECTIVETo compare the antitussive activity of three kinds of Stemonae Radix specified in the Chinese Pharmacopoeia, including roots of Stemona sessilifolia, S. japonica and S. tuberosa.

METHODThe antitussive activity was determined in mouse after cough induction by ammonia aerosol stimulation and the number of cough in 2 min were detected with codeine as positive control.

RESULTAll the decoctions, the total alkaloid fractions and non-alkaloid fractions of S. sessilifolia, S. japonica and three chemical types of S. tuberosa showed significant antitussive effect and exhibited a dose-dependent inhibition of coughing. The ED50 values showed that the antitussive activity strength for both total alkaloid fractions and the decoctions are: S. tuberosa (Type I) > S. sessilifolia > S. japonica. The total alkaloid fractions had more potent atitussive activity than the decoctions and non-alkaloid fractions. The antitussive activity strength for the three chemical types of S. tuberosa is: Type I > Type III > Type II. The samples from different producing areas for the same species of Stemonae Radix had no significant differences in antitussive activity. The result also showed that the honey-processed slice had much stronger antitussive activity than raw slice.

CONCLUSIONThe antitussive efficacies of Stemonae Radix were influenced by chemical diversity both in same species and among different species, different fractions and processed method.

Animals ; Antitussive Agents ; administration & dosage ; Cough ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Stemonaceae ; chemistry

Objective:To investigate the diagnostic value of neutrophil CD64 index (nCD64) in disseminated nontuberculous mycobacteria (NTM) infection.Methods:Thirty-six patients with NTM infection from January 2020 to June 2021 in Huashan Hospital, Fudan University were included. Patients were classified into groups of disseminated infection and focal infection according to their medical history and discharge diagnosis. The expressions of nCD64 in patients with focal infection and disseminated infection before treatment were collected and analyzed. Statistical analysis was performed using the Mann-Whitney U test, and the diagnostic value of nCD64 for disseminated NTM infection was analyzed using the receiver operator characteristic curve (ROC curve). Results:Among the 36 patients with NTM infection, 18 cases were focal infection (due to the low white blood cell count of the patient with myelodysplastic syndrome, the detection results were biased, which were excluded from the subsequent analysis) and 18 cases were disseminated infection. The expression of nCD64 in focal infection was 0.72(0.50, 1.55), and that in disseminated infection was 13.63(6.77, 32.31). The difference was statistically significant ( U=15.50, P<0.001). Using focal infection as a control, the area under the ROC curve for the operational characteristics of the subjects was 0.949 3 for disseminated NTM infection. The diagnostic cut-off value of nCD64 was 3.06, with the sensitivity and specificity of the disseminated NTM infection were 88.89% and 100.00%, respectively. Conclusions:In patients with NTM infection before effective treatment, the diagnostic cut-off value of nCD64 of 3.06 has high sensitivity and specificity, which is useful for the aided diagnosis of disseminated NTM infection.

Objective:To explore the expression and clinical significance of immunosuppressive receptor T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) on the peripheral blood mononuclear cells (PBMC) in silicosis patients with Mycobacterium tuberculosis infection. Methods:August 2018, a total of 78 patients with silicosis (all were quarry workers in Sanmen County, Zhejiang Province) were enrolled and divided into silicosis combined with active pulmonary tuberculosis group (APTB group), silicosis combined with latent tuberculosis infection group (LTBI group), and simple silicosis with non-tuberculosis infection group (non-TB group). Flow cytometry was used to analyze the expressions of TIGIT, programmed death-1 (PD-1) and transcription factor T-bet on PBMC from patients. Mann-Whitney U test and Pearson correlations analysis were used for statistical analysis. Results:Among the 78 patients, eight were in the APTB group, 24 in the LTBI group, and 46 in the non-TB group. The expressions of PD-1 and TIGIT on CD8 + T cells in the APTB group (29.45%(16.78%) and 65.40%(12.12%), respectively) were significantly higher than those in the LTBI group (17.40%(11.17%) and 48.30%(28.75%), respectively; U=23.500 and 43.500, respectively, P=0.000 8 and 0.020 5, respectively) and non-TB group (15.95%(12.46%) and 45.30%(19.75%), respectively; U=64.000 and 69.000, respectively, P=0.002 3 and 0.003 8, respectively), and the differences were all statistically significant. The expression of TIGIT was positively correlated with PD-1 on CD8 + T cells in silicosis patients ( r=0.434 3, P<0.01). The proportion of PD-1 + TIGIT + CD8 + T cells in the APTB group (19.90%(22.67%)) was significantly higher than those in the non-TB group (11.55%(11.29%), U=76.500, P=0.007 1) and LTBI group (11.55%(10.53%), U=41.000, P=0.015 4), while the proportion of PD-1 -TIGIT -CD8 + T cells in the APTB group (30.60%(12.90%)) was significantly lower than non-TB group (48.90%(18.98%), U=58.000, P=0.001 3) and LTBI group (47.20%(24.59%), U=41.000, P=0.015 4). The differences were all statistically significant. The expression of T-bet on the peripheral blood CD8 + T cells in the APTB group (29.45%(16.78%)) was higher than that in the non-TB group (15.95%(12.46%)) and the LTBI group (17.40%(11.17%)), and the differences were both statistically significant ( U=46.500 and 46.000, respectively, P=0.000 3 and 0.028 3, respectively). The expression of T-bet on CD8 + T cells was positively correlated with TIGIT on CD8 + T cells ( r=0.456 7, P<0.01). The expression of T-bet on PD-1 + TIGIT + CD8 + T cells in the APTB group (65.40%(12.12%)) was higher than those in the LTBI group (48.30%(28.75%), U=23.500, P=0.000 8) and non-TB group (45.30%(19.75%), U=65.000, P=0.002 6), and the differences were both statistically significant. Conclusion:The immunosuppressive receptor PD-1 and TIGIT are highly expressed on CD8 + T cells in silicosis patients with active pulmonary tuberculosis, which indicates CD8 + T cells exhaustion in these population, while the highly co-expression of T-bet suggests the exhausted subsets may have reversed potentiality.

To effectively screen treating pigmentation disorders drug, a new transgenic zebrafish drug screening model was constructed. Green fluorescent protein(GFP)were drived by tyrosinase-related protein 1a (tyrp1a)promoter specific expression in melanocyte. Effect of N-Phenylthiourea(PTU)and alpha-melanaocyte stimulating hormone(α-MSH)on the GFP expression and melanogenic of tyrp1a: eGFP zebrafish were photographed under the steromicroscope. Results showed that PTU could significantly inhibit the melanogenic and expression of GFP of zebrafish. As compared with control group, α-MSH treatment resulted in marked stimulation of body pigmentation and expression of GFP. In conclusion, a new transgenic zebrafish drug screening model was successfully established, which can be used for treating pigmentation disorders.

Objective To retrospectively analyze and compare the clinical efficacy and safety between fractionated stereotactic radiotherapy (FSRT) combined with and without temozolomide in the treatment of large brain metastases.Methods Between 2009 and 2017,84 patients with large brain metastases (tumor size ≥ 6 cm3) were recruited and assigned into the CRT group (concurrent TMZ and FSRT,n=42) and RT group (FSRT alone,n=42).The radiation dose was 52.0 Gy in 13 fractions or 52.5 Gy in 15 fractions.Patients were reexamined by magnetic resonance imaging (MRI) during treatment.The radiation field would be shrunk if the gross target volume (GTV) was reduced.The clinical efficacy was evaluated at postoperative 2 to 3 months.The primary end-point event was local recurrence-free survival (LRFS) and the secondary end-point events included intracranial progression-free survival (IPFS),progression-free survival (PFS),overall survival (OS),brain metastasis-specific survival (BMSS) and adverse events.The survival rates were assessed with Kaplan-Meier method and log-rank test and monovariate analysis.Results The median GTV in the CRT and RT groups was 16.9 cm3 and 15.7 cm3.During the treatment,75％ of the lesions in the CRT group were reduced compared with 34％ in the RT group (P=0.000).The local control (LC) rate in the CRT and RT groups was 100％ and 98％.The median follow-up time was 16.1 months (range,2.1-105.7 months).In the CRT group,the LRFS (P=0.040),IPFS (P=0.022),PFS (P=0.045),OS (P=0.013) and BMSS (P=0.006) were significantly better than those in the RT group,respectively.In the CRT group,the incidence of grade Ⅰ-Ⅱ gastrointestinal adverse events was 33％,significantly higher compared with 26％ in the RT group (P=0.006).No grade Ⅳ-Ⅴ adverse events occurred in both groups.Conclusion Combined application of temozolomide and FSRT can further enhance the LC and survival rates and do not increase the risk of severe adverse events in patients diagnosed with large brain metastases.

Objective To retrospectively analyze the dosimetry and efficacy of whole-brain irradiation (WBRT) with simultaneous integrated boost (SIB) by helical tomotherapy (HT) in the treatment of multiple brain metastases (BMs),and to evaluate the feasibility,efficacy,and safety of HT.Methods From 2014 to 2017,a total of 43 patients with multiple BMs (no less than 3 lesions) were enrolled as subjects.A dose of 40 Gy was delivered to the whole brain in 20 fractions,while a dose of 60 Gy was delivered to the gross target volume (GTV) in 20 fractions.Patients were reexamined by magnetic resonance imaging during treatment.The radiation field would be shrunk if GTV was reduced.Target coverage (TC),conformity index (CI),prescription isodose/target volume (PITV) ratio,and homogeneity index (HI) were assessed.Clinical indices included local recurrence-free survival (LRFS),intracranial progression-free survival (IPFS),progression-free survival (PFS),overall survival (OS),and toxicities.Results The median lesion number was 6(3-36) and the median total volume of GTV was 8.74 cm3.The TC,CI,PITV,and HI for GTV were 0.96±0.028,0.51±0.164,2.09±1.245,and 0.12±0.066,respectively,while the TC and HI for the whole brain were 0.95±0.033 and 0.43±0.161,respectively.In all the patients,26％ had replarming during treatment.The two-stage treatment reduced the radiation dose to organs at risk.The 1-year LRFS,IPFS,PFS,and OS rates were 96％,80％,39％,and 86％,respectively.No grade ≥3 toxicities were observed.Conclusions WBRT with SIB by HT achieves satisfactory conformity,homogeneity,efficacy,and safety,which is a recommended treatment plan for multiple BMs.Replanning during treatment can better protect normal tissue.

Objectives To evaluate the clinical efficacy and safety of hypofractionated radiotherapy for cancer patients with hepatic metastases. Methods From May 2007 to November 2016,45 patients ( male:female=20:25) with inoperable hepatic metastases were enrolled in this investigation. The median age was 58 years old ( range:25-83).The median Karnofsky performance score ( KPS) was 80.Primary colorectal cancer was detected in 14 patients,primary breast cancer in 9 and primary lung cancer in 6 cases. Twenty-one patients had extrahepatic metastases. A total of 52 lesions were treated. Thirty-four cases received radiotherapy for one single lesion. The fractional dose was 45 Gy/3 fractions and 60 Gy/10-15 fractions. The median gross tumor volume (GTV) was 10. 1 cm3(0. 3-175. 2 cm3) and 29. 8 cm3(5. 0-209. 6 cm3) for planning target volume ( PTV).Seventeen CT images were fused with MRI and IMRT was adopted in 43 cases. The median dose of PTV was 60 Gy (40-60 Gy) and 90 Gy (60-132 Gy) for bioequivalent dose (BED). Results The median follow-up time was 23. 5 months and the median survival time was 26. 0 months (95%CI:21.4-30.6 months).The 1-year local control (LC),disease-free survival (DFS) and overall survival ( OS ) were 94%, 27% and 91%, respectively. Six cases died of liver metastases and abnormal liver function. Conclusion Hypofractionated radiotherapy is an efficacious and safe local treatment for inoperable hepatic metastases.

Objective:To analyze the differences of peripheral blood transcriptome between mild and severe influenza A (H1N1) patients, and to find indicators for the assessment of disease severity.Methods:A total of ten patients (five patients with mild disease and five patients with severe disease) diagnosed with H1N1 infection from January to May 2018 at Huashan Hospital, Fudan University in Shanghai were enrolled, and five healthy people were also enrolled as controls. The peripheral blood of patients was collected for transcriptome sequencing at the time when they were first diagnosed. Measurement data were compared using t test or Mann-Whitney U test. The count data were compared using Fisher exact test when appropriate. Data analysis of transcriptome predictions was performed using bioinformatics methods. Results:The platelet counts were significantly different between mild and severe groups ((163.4±21.5 )×10 9/L vs (255.6±52.5)×10 9/L, t=3.636, P=0.007). There were no differences between the two groups in gender, age, white blood cell counts, neutrophil percentage, lymphocyte percentage and hemoglobin levels (all P>0.05). However, the average expression levels of matrix metalloproteinase (MMP) 8 and MMP9 in severe group (18.41 and 174.00, respectively) were both higher than those in mild group (2.33 and 22.91, respectively) and healthy control (1.43 and 34.65, respectively; all P<0.01). Conclusion:MMP8 and MMP9 could be expected to serve as the molecular biological markers for predicting the disease severity in patients with influenza A (H1N1) infection.