To improve the status of management in respiratory failure in China,the project of Study on Pathogenesis and Treatment of Respiratory Failure was designed and conducted by three medical centers(Beijing Institute of Respiratory Medicine-Beijing Chaoyang Hospital,Affiliated to Capital Medical University,Zhongshang Hospital-Fudan University,Guangzhou Institute of Respiratory Medicine-First Guangzhou Medical College)for more than ten years.This project was focused on pathogenesis and treatment strategies of respiratory failure and achieved the following important innovations:(1)Pulmonary Infection Control Window(PIC Window)was firstly proposed and used to determine the time switching point of sequential invasive-noninvasive ventilation;(2)The largest sample size of early use of noninvasive positive pressure ventilation(NPPV)for acute exacerbated COPD(AECOPD)on general ward provided the evidence-based data for expanding the indication of NPPV from treating respiratory failure to alleviating respiratory muscle fatigue;(3)Three new types of masks with intellectual property for NPPV were developed;(4)Designing of intrinsic expiratory end positive pressure(PEEPi)lung model with property of expiratory flow limitation confirmed that PEEPi was the most important factor that increased inspiratory difficulty;(5)The systematic measurement was established for diaphragm strength and endurance;(6)Aquaporin 1(AQP1)was firstly proved the key channel of fluid transportation in the lung;(7)A multicenter prospective cohort study provided objective data that depression had causal effect on COPD exacerbation and hospitalization;(8)Two guidelines for NPPV and mechanical ventilation of AECOPD were initiated by this group.This project has been widely used in clinical practice and promoted the research and treatment of respiratory failure in China.

Objective To investigate the possibility of human papillomavirus late 1 (HPV-L1) protein detection in predicting the prognosis of cervical intraepithelial neoplasia (CIN). Methods (1)Through immunocytochemical method to detect expression of HPV-L1 protein in diagnosis of CIN before treatment. (2) Through hybridization and gene chip technology to detect expression of high-risk human papilloma virus (HR-HPV) in diagnosis of CIN after treatment for twelve months. Results (1) Comparing the three treatment methods in CIN patients, HR-HPV infection sustained no significant difference (P > 0.05). (2) The positive expression rate of HPV-L1 protein was 75.00% in CINⅠand 36.13%in CINⅡ~Ⅲbefore treatment, with statistically significant (P<0.05). (3) Persistent infection rate of HR-HPV was 5.97%in HPV-L1 positive expression group after treatment for twelve months and 19%in HPV-L1 negative expression group. The difference between the two was statistically significant (P<0.05). (4) The expression of HPV-L1 capsid protein before treatment had a negative relationship with HR-HPV persistent infection after 12 months of treatment. Conclusion Detection of HPV-L1 protein before treatment can predict the prognosis of CIN which had been treated.

Objective To develop a suitable extraction and purification method for detecting the residue of 20 kinds of organochlorine pesticides(OCPs) in suppositories of Chinese medicinal materials which are made of fat-soluble bases.Methods Different methods were applied for the extraction, and GPC was used for the purification of the sample Huazhi Shuan, and then the residue contents of OCPs were determined by GC- ECD method.20 kinds of OCPs were added to the samples and the feasibilities of the methods were evaluated by the obtained recovery. The established method were used in the analysis of the residue of OCPs in other Chinese herbal suppositories.Results The recovery and the reproducibility of this method met the requirements for the analysis of pesticides residues. Conclusion This developed method may provide reference for the detection of organochlorine pesticides residue in these kinds of Chinese medicinal preparations.

Objective To develop a suitable solid-phase extraction(SPE)method for the further purification of the test solu- tion in suppositories of Chinese medicinal materials which are made of lipid soluble bases,and to detect the residues of 20 kinds of organochlorine pesticides(OCPs).Methods Different kinds of SPE columns were selected and several elution solvents were used for the purification of the sample of herbal medicine of Huazhi Shuan,and then the residue contents of OCPs were determined by GC-ECD method.Twenty kinds of OCPs were added into the samples and the efficacy of the methods were eval- uated by the obtained recoveries.Results The recoveries of the 20 kinds of OCPs being detected were basically comply with the requirements for the analysis of pestiside residues.Conclusion After the tested solution is obtained from GPC column and is further purified through florisil SPE column,the impurity can be removed and the high accuracy of the quantitative analysis of the compounds can be achieved.

Tumors represent one of the primary threats to human life, with the dissemination of malignant tumors being a leading cause of mortality among cancer patients. Early diagnosis of tumors can reliably predict their progression, significantly reducing mortality rates. Tumor markers, including circulating tumor cells, exosomes, proteins, circulating tumor DNA, miRNAs and so on, generated during the tumor development process, have emerged as effective approach for early tumor diagnosis. Several methods are currently employed to detect tumor markers, such as polymerase chain reaction, Northern blotting, next-generation sequencing, flow cytometry, and enzyme-linked immunosorbent assay. However, these methods often suffer from time-consuming process, high costs, low sensitivity, and the requirement for specialized personnel. Therefore, a new rapid, sensitive, and specific tumor detection method is urgently needed.The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system, originating from the adaptive immune system of bacteria, has found extensive applications in gene editing and nucleic acid detection. Based on the structure and function of Cas proteins, the CRISPR/Cas system can be classified into two classes and six types. Class I systems consist of multiple Cas protein complexes, including types I, III, and IV, while Class II systems comprise single, multi-domain Cas proteins mediated by RNA, including types II (Cas9), V (Cas12), and VI (Cas13). Class II systems have been widely employed in the fields of biotechnology and nucleic acid diagnostics due to their efficient target binding and programmable RNA specificity. Currently, fluorescence method is the most common signal output technique in CRISPR/Cas-based biosensors. However, this method often requires the integration of signal amplification technologies to enhance sensitivity and involves expensive and complex fluorescence detectors. To enhance the detection performance of CRISPR/Cas-based biosensors, the integration of CRISPR/Cas with some alternative techniques can be considered. The CRISPR/Cas integrated electrochemical sensor (E-CRISPR) possesses advantages such as miniaturization, high sensitivity, high specificity, and fast response speed.E-CRISPR can convert the reactions between biomolecules and detecting components into electrical signals, rendering the detection signals more easily readable and reducing the impact of background values. Therefore,E-CRISPR enhances the accuracy of detection results. E-CRISPR has been applied in various fields, including medical and health, environmental monitoring, and food safety. Furthermore, E-CRISPR holds tremendous potential for advancing the detection levels of tumor markers.Among all types of Cas enzymes, the three most widely applied are Cas9, Cas12, and Cas13, along with their respective subtypes. In this work, we provided a brief overview of the principles and characteristics of Class II CRISPR/Cas single-effector proteins. This paper focused on the various detection technologies based on E-CRISPR technique, including electrochemical impedance spectroscopy, voltammetry, photoelectrochemistry, and electrochemiluminescence. We also emphasized the applications of E-CRISPR in the field of tumor diagnosis, which mainly encompasses the detection of three typical tumor markers (ctDNA, miRNA, and proteins). Finally, we discussed the advantages and limitations of E-CRISPR, current challenges, and future development prospects. In summary, althoughE-CRISPR platform has made significant strides in tumor detection, certain challenges still need to be overcome for their widespread clinical application. Continuous optimization of the E-CRISPR platform holds the promise of achieving more accurate tumor subtyping diagnoses in clinical settings, which would be of significant importance for early patient diagnosis and prognosis assessment.

Aim To develop a simple and rapid method to monitor insulin receptor kinase activity and provide a novel cell-based model for screening anti-diabetes drugs.Methods CHO cells were co-transfected by plasmids which respectively contained insulin receptor gene,STAT5b gene and luciferase gene driven by STAT5 response elements.The expression of exogenous gene in transfected cells was examined by RT-PCR.The transfected cells were treated by insulin,and then the concentration and time-dependent response of luciferase expression to insulin induction was examined.Moreover,the specificity was identified by AG1024 treatment and PTP1B gene transfection.Results Expressions of insulin receptor and STAT5b were detected in the transfected CHO cells.The expression of luciferase in transfected cells was induced by insulin in concentration and time-dependent way.The maximal induction fold was 6.25.Moreover,the inducible expression of luciferase by insulin could be specifically blocked by tyrphostin AG1024,an inhibitor of insulin receptor kinase,or co-transfected PTP1B gene.Conclusions The insulin receptor kinase activity can be detected by expression of reporter gene with high sensitivity and specificity in this cell model,and with potential value in high throughput screening for insulin receptor activators and sensitizers.

Stable isotope coded derivatization (ICD) is an isotope labeling technique for specific functional groups of the target analytes through chemical derivatization.ICD combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) enables systematically analyzing the biomolecules with the same labeling reaction characteristic.ICD technique can effectively solve the limited sensitivity in complex bio-matrices analysis and unavailability of isotope internal standards in quantitative analysis.In recent years, ICD technique has been widely used in metabolomics research field.Based on this background, the ICD technique and the design of ICD reagents are briefly described in this review.The recent advances in ICD reagents for carboxyl, amino, carbonyl, thiol and hydroxyl groups and their applications in the analysis of small molecule metabolites in bio-matrices with LC-MS/MS are reviewed.

Objectives: To explore the role of liver in the process of severe acute pancreatitis. Methods:Comparing the survival time, the endotoxin level of plasma and ascites , the IL-6 level of serum and ascites, the platelet granule membrane protein-140 (GMP-140) level of plasma and the histology change of lung between control group, portocaval shunting group(PC), acute necrotic pancreatitis (ANP) group and acute necrotic pancreatitis immediately after portocaval shunt(PC+ANP) group of rats. Measuring the serum IL-6 of control group, portocaval shunt control group (injecting normal saline through caval vein, PCJ), ascites injecting group (AJ) and ascites injecting portocaval shunt group (PC+AJ). Results: The survival time of PC+ANP group was much shorter than those of the other groups, and its plasma endotoxin , serum IL-6 and plasma GMP-140 levels were higher than those of the other groups (P

Objective To investigate the association between morniing blood pressure surge (MBPS) and carotid atherosclerosis in elder patients with essential hypertension. Methods According to the results of24h ambulatory blood pressure monitoring, 106 patients were classified as the morning BP surge group (MBPS group,n = 58) ,and nonsurge group (NMBPS group, n = 48). Patients underwent carotid ultrasound and the intima-medial thickness (CCA-IMT) and plaques were examined. Results The CCA-IMT of the MBPS group was significantly thicker than that the NMBPS group[(1.27 ± 0. 12)mm vs (0.92 ± 0.33 )mm], P ＜ 0. 05 ) ;②Compared with the NMBPS group,the severity of carotid arteries plaque of the MBPS group was significantly higher (72. 15% vs 54.21% ), ( P ＜0. 01 ) ;③Pearson relation analysis showed CCA-IMT level positively correlated with age (r = 0.288, P ＜ 0.001 ) ,the average of 24h SBP ( r = 0. 768 ,P ＜ 0. 001 ), and MBPS ( r = 0. 768, P ＜ 0.001 ). Conclusion The study showed that MBPS was closely related with carotid atherosclerosis in elder patients with essential hypertension and was an important risk factor in the process of atheresclerosis.