Single-cell analysis reveals dysregulated inflammatory response in peripheral blood immunity in patients with acute respiratory distress syndrome.

Introduction: Acute respiratory distress syndrome (ARDS) remains a major clinical challenge for patients in intensive care units. Determining the differential mechanisms underlying ARDS with different etiologies is a key goal to improve the effectiveness of ARDS therapy. Despite growing evidence that different immune cell types are involved in ARDS, the role of altered immune cell subpopulations in disease progression is unelucidated. Methods: In this study, we combined scRNA-seq and bulk-level sequencing to analyze the transcriptomes of peripheral blood mononuclear cells from healthy volunteers and patients with septic ARDS (sep-ARDS) and pneumonic ARDS (PNE-ARDS). Results: Our data revealed differential alterations at the cellular and molecular levels and within biological signaling pathways in ARDS with different etiologies. The dynamics of neutrophils, macrophages (Macs), classical dendritic cells (cDCs), myeloid-derived suppressive cells (MDSCs), and CD8+ T cells varied significantly among groups of different samples, with neutrophils and cDCs at higher, and Macs at significantly lower, amounts in the patients with sep-ARDS. Furthermore, MDSCs were highly enriched only in the sep-ARDS patients, whereas a higher abundance of CD8+ T cells was observed in patients with PNE-ARDS. In addition, these cell subpopulations were found to be significantly involved in apoptosis, inflammatory, and immune-related pathways. In particular, a significant enhancement of the oxidative stress response was observed in the neutrophil subpopulation. Conclusion: Our study shows that the composition of cells involved in the main peripheral circulation differs in patients with ARDS with different etiologies. Studying the role and mechanism of action of these cells during ARDS will provide new opportunities for the treatment of this condition.

Serum profile of low molecular weight fucosylated glycoproteins for early diagnosis of hepatocellular carcinoma.

Our previous study reported a method of using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to analyze the association between abnormal fucosylation of serum glycoproteins and the progression of hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC). In the present study, the aforementioned method was improved by focusing on fucosylated glycoproteins <10 kD, classification models were established and blind tests were performed on an enlarged sample size (n=299). According to the present results, the classification models had a sensitivity and specificity of 74.31 and 76.32%, respectively, to identify HCC among all serum samples, 81.65 and 83.08%, respectively, to distinguish HCC from HBV-associated cirrhosis and chronic hepatitis Band 88.99 and 84.62%, respectively, to distinguish HCC from HBV-associated cirrhosis. When combined with α-fetoprotein (AFP) measurements (AFP >20 ng/ml), the sensitivity and specificity of the models were significantly elevated to 80.73 and 87.37%, 87.16 and 90.00%, and 92.66 and 93.84%, respectively. In addition, the HBV-HCC vs. HBV-cirrhosis classification model was used to analyze serum samples collected from 9 patients with cirrhosis 1 year before they were diagnosed with HCC, and from 6 patients who had cirrhosis but developed no signs of HCC for the following 3 years. The model identified 7 patients (77.78%) with no significant clinical symptoms of HCC, and gave no false positive results, demonstrating that the classification models established in the present study may be useful for the early diagnosis of HCC. After isolation and purification, two proteins with differential expression were identified as isoform 1 of inter-α-trypsin inhibitor heavy chain 4 precursor, and thymosin ß-4-like protein 3. These may be used as candidate markers for HCC diagnosis. Additionally, the present study indicates that defucosylation of serum glycoproteins may occur during the development and progression of HCC.

Formation of Nϵ-Carboxymethyllysine and Nϵ-Carboxyethyllysine in Prepared Chicken Breast by Pan Frying.

The objective of this work was to investigate the effects of pan frying on the formation of two typical advanced glycation end products (AGEs) named Nϵ-carboxymethyllysine (CML) and Nϵ-carboxyethyllysine (CEL) in prepared chicken breast. The marinated chicken breast was pan fried for 1 to 6 min, and then protein, fat, moisture, carbonyl, sulfhydryl, thiobarbituric acid reactive substances, chroma (a*, b*, L*), absorbance at 294 and 420 nm, and AGE (CEL) levels were measured once a minute. Pearson's correlation was determined and indicated that moisture was significantly negatively correlated with Maillard reaction and AGEs (P < 0.05), fat and protein contents were significantly positively correlated with AGEs (P < 0.05), and a* values were positively correlated with Maillard reaction and CEL (P < 0.05). Protein and lipid oxidation played an important role on the correlation of AGEs. In conclusion, Maillard reaction and oxidation reaction are two important factors affecting AGE formation.

Development of a novel ssDNA aptamer targeting neutrophil gelatinase-associated lipocalin and its application in clinical trials.

BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is a promising biomarker of early diagnosis and prediction for acute kidney injury (AKI). However, the current program for NGAL detection is not extensively applied in clinics due to the high expense of antibodies. Nucleic acid aptamers are single-strand DNAs or RNAs which could bind to targets with high specificity and affinity, and they have been widely used in the diagnosis and therapy for multiple diseases. It is valuable for us to develop a new method for NGAL detection using aptamers instead of antibodies to achieve increased efficiency and decreased cost. METHODS: Nucleic acid aptamers against NGAL were obtained after SELEX process using magnetic beads, and an enzyme-linked aptamer analysis (ELAA), which can be widely used in clinical diagnosis at low cost, were successfully established. The feasibility of ELAA was further validated with urine samples harvested from 43 AKI patients and 30 healthy people. RESULTS: Three candidate aptamers, including NA36, NA42 and NA53, were obtained after 8 rounds of SELEX process with magnetic beads and verified by quantitative polymerase chain reaction (qPCR), and the Kd value of each aptamer was 43.59, 66.55 and 32.52 nM, respectively. Moreover, the linear relationship was consistent at the range of 125-4000 ng/mL, and the detection limit of ELAA assay was 30.45 ng/mL. We also found that NGAL could be exclusively detected with NA53, and no cross-reaction between NA53 and human albumin or globulin occurred, the coefficient of variation (CV) between inner-plate and inter-plate was less than 15%, and the recovery rate was between 80 and 110%. Moreover, the sensitivity and specificity of ELAA assay in this study are 100% and 90%, respectively. Consistently, these results could also diagnose whether the occurrence of AKI in lots of patients, which has been demonstrated with the ELAA method we established after using NA53. CONCLUSIONS: Taken together, NA53, the best candidate aptamer targeting NGAL protein, can be applied in clinical testing.

[Clinical effect on the treatment of the low-middle frequency sudden hearing loss with postaurical injection of methylprednisolone].

OBJECTIVE: To evaluate the clinical efficacy on the treatment of the low-middle frequency sudden hearing loss with postaurical injection of methylprednisolone. METHOD: The 80 cases of the low-middle frequency sudden hearing loss were randomly divided into postaurical injection and oral hormone groups. The postaurical injection group (42 cases) received the postaurical injection of methylprednisolone, 40 mg/2 d, combined with the treatment of Ginkgo dipyidamolum and Alprostadil for 14 d; The oral hormone group (38 cases) received the oral prednisone, 1 mg/kg/d, administrated once on the morning for 3 d, if effective, prolonging for another 2 d, as mentioned above for Ginkgo dipyidamolum and Alprostadil. RESULT: The total effective rate was 88.10% in postaurical injection group and 86. 4o%in oral hormone group. There was no significant difference between the twbogroups( P> 0. 5). CONCLUSION: Postaurical injection of methylprednisolone for the low-middle frequency sudden hearing loss is effective, safe and simple, which may be an alternative for systemic administration of gulcocorticoid.

[Evaluation the curative effect of supracricoid partial laryngectomy for laryngeal cancer].

OBJECTIVE: To analyze the oncological outcomes, functional outcomes in patients undergoing supracricoid partial laryngectomy (SCPL). Provide clinical experience for application of SCPL. METHOD: A retrospective analysis of the 115 cases with laryngeal carcinoma accepted SCPL in our department from Jan 1996 to Dec 2004. Use the Kaplan-Meier method to analyze the patients'survival rate. Evaluate the value of reserve larynx function. RESULT: The 5-years survival rates and the decannulation rate was 80.8%, 99.1% respectively; and the average decannulation time was 22.25 days. The mean time of removal of gastric tube was 9.57 days. The function of CHEP was superior to CHP. The vocal function of 115 cases were all achieved in general communication. CONCLUSION: SCPL get better oncological and functional outcomes and allows the preservation of the basic function of the larynx. It's a safe, effective technique and deserved to generalization.

[Evaluation of effectiveness of advanced tonsillar carcinoma by different treatment].

OBJECTIVE: To evaluate the treatment of advanced tonsillar carcinoma by radiotherapy plus salvage surgery (R+S) or surgery coupling with postoperative radiotherapy (S+R). METHOD: Clinical data of 48 patients with advanced tonsillar carcinoma who were treated in The 2nd Affiliated Hospital of Sun Yat-sen University from June 1996 to June 2004 was retrospectively analyzed. The patients were divided into R+S group (group A, 21 cases) and S+R group (group B, 27 cases). Treatment outcomes were compared between these two groups. The QOL (quality of life) scale of Washington University (UW-QOL) was used to investigate the patient's quality of life. RESULT: The 5-year survival rates were 42.9% in group A and 45.8% in group B, there was no significant statistical difference between the two groups (P < 0.05). Both the two treatment modalities could reduce the QOL in some degree. The average QOL score of 46 patients was 661.00 +/- 98.52 , group A was 696.09 + 90.70, while group B was 631.52 +/- 96.74, there was a significant statistical difference between the two groups (P < 0.05). CONCLUSION: The two treatment modalities reached similar survivals. However, compared with the S+R, some patients who accepted treatment of R+S could avoid composite resection, reduce functional lesion and improve the QOL.