With the organic combination of rapid on-site evaluation (ROSE) and interventional pulmonary diagnostic technology, we can build a complete The System of Diagnostic Interventional Pulmonology Based on Rapid on-site Evaluation. With the help of ROSE, changing the ways, methods and modalities of interventional pulmonary diagnostic technology to obtain the target lesions is the core of this system. In this statement, the most commonly used standard operating techniques in The System of Diagnostic Interventional Pulmonology Based on Rapid on-site Evaluation are described in detail, including double-hinge curette operating technique, transbronchial lung biopsy (TBLB) technique, and transbronchial brushing technique.

Abstract: Objective　To investigate the distribution and drug resistance of pathogenic bacteria separated from ascites of patients in Children’s Hospital Affiliated to Zhengzhou University from 2015 to 2021, and to provide a basis for rational clinical antimicrobial agents. Methods　Bacterial culture, bacterial identification and drug sensitivity analysis were performed on 1 058 non-duplicate ascites culture specimens from January 2015 to December 2021. The clinica1 and microbiologica1 data were ana1yzed by WHONET 5.6 and SAS 9.4 Results　Of the 1 058 specimens, 586 (55.39%) were positive for pathogenic bacteria, with a total of 781 strains isolated. There was no significant trend of increase or decrease in the positivity rate over different years. Male children (63.99%) were more prevalent than female children. Appendicitis (59.22%) was the most common disease and Escherichia coli was the most common causative bacteria. Among neonates (≤28 d), the bacteria with the highest detection rate were Klebsiella pneumoniae (23.50%) and Enterococcus faecium (23.50%), while among children (>28 d), the highest detection rate was Escherichia coli (35.98%). Gram-negative bacteria accounted for 64.79% of the 781 strains, mainly Escherichia coli (38.28%), Klebsiella pneumoniae (8.58%), and Pseudomonas aeruginosa (5.89%); Gram-positive bacteria accounted for 29.45%, mainly Enterococcus faecium (8.58%), Streptococcus constellatus (2.69%), and Enterococcus avium (2.43%); fungi accounted for 1.66% and anaerobic bacteria accounted for 4.10%. The resistance rates of Escherichia coli to cefoperazone/sulbactam, piperacillin/tazobactam, imipenem and meropenem were 6.02%, 4.35%, 4.35%, and 3.68%, respectively. The resistance rates of Klebsiella pneumoniae to these drugs were 59.70%, 59.70%, 50.75% and 53.73% respectively. Linezolid-resistant strains of Enterococcus faecium were found. Conclusion　Appendicitis is the most common abdominal infection in children, and the distribution of ascites pathogens varies with ages and diseases. The pathogenic bacteria are mainly Gram-negative bacteria, and the drug resistance of Klebsiella pneumoniae was more serious. It is particularly important to use antibiotics correctly and rationally to reduce the emergence of drug resistant bacteria.

Objective To explore the anatomic character of brachial plexus nerve by ultrasonography,and provide a basis for ultrasound-guided brachial plexus block.Methods Twenty-four healthy volunteers were chosen for high-frequency sonography examination,the features of brachial plexus distributions were studied in the axilla location and the interscalene location.Results Brachial plexus of all the 24 cases were detected.At the axilla level,main braches of brachial plexus nerves were very close to the axillary artery.At the interscalene level,the brachial plexus was consistently found between the scalenus anterior and medius muscles in the interscalene groove.Conclusions Using high-frequency sonography,the main braches of brachial plexus can be observed clearly.

Aim To investigate the change of miR-124 expression in methamphetamine-induced addiction in PC12 cells and the possible regulatory mechanism that it involves.Methods PC12 cells were randomly divided into 6 groups as follows: control group, methamphetamine group, agomir Negative Control group, miR-124 agomir group, agomir Negative Control+methamphetamine group and miR-124 agomir+methamphetamine group.After the treatment, the total RNA and protein were extracted in PC12 cells.The expression of miR-124 was measured by Real-time PCR and the expression of GluR2 was determined by Western blot in PC12 cells.Results Compared with those in the control group, the expression of miR-124 was remarkably decreased and the expression of GluR2 was significantly increased in the methamphetamine group in PC12 cells.Compared with those in the agomir Negative Control+methamphetamine group, the expression of miR-124 was remarkably increased and the expression of GluR2 was significantly decreased in the miR-124 agomir+methamphetamine group in PC12 cells.Conclusion MiR-124 might involve in methamphetamine-induced addiction in PC12 cells by inhibiting GluR2.

Objective To analyze the medication rules of the treatment of stroke in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan; To provide references for the clinical treatment. Methods Prescriptions with confirmed efficacy of famous TCM doctors in the history in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan was searched. Excel2003 was used to establish database to analyze medication frequency. SPSS17.0 statistical software was used to conduct cluster analysis, and tree view was used to show results. Results Totally 112 prescriptions for the treatment of stroke of 36 famous TCM doctors were included in the study, including 204 kinds of Chinese materia medica and 1169 times of medication frequency. The high-frequency medicines (>10 times) were Paeoniae Radix Alba, Achyranthis Bidentatae Radix, Angelicae Sinensis Radix, Pinelliae Rhizoma Praeparatum, and Poria. The high-frequency medicine categories were tonifying deficiency medicine (22.58％), pacifying liver and wind medicine (12.31％), activating blood and dispelling stasis medicine (11.89％), clearing heat medicine (11.46％) and dissipating phlegm, cough and asthma medicine (8.72％). Cluster analysis showed that high-frequency medicine (>10 times) could be clustered as 6 categories. Conclusion Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan focuses on tonifying deficiency medicine, accompanied with pacifying liver and wind medicine, activating blood and dispelling stasis medicine, clearing heat medicine and dissipating phlegm, cough and asthma medicine, which can be used to guide clinic.

Objective To explore the effects and mechanisms of hepatitis B virus-X protein (HBx) on invasion and migration of hepatocellular carcinoma (HCC) cells.Methods The retrospective cohort study was conducted.The clinicopathological data of 30 patients with liver tumor (20 with HCC and 10 with benign tumor of liver) who were admitted to the Affiliated Hospital of Xuzhou Medical College between July 2014 and July 2015 were collected.HCC tissues of 20 patients with HCC (with history of HBV infection) were collected by surgical resection and peritumoral normal tissues (outside of tumor capsule) of 10 patients with benign tumor of liver (without history of HBV infection) were collected.The expressions of epidermal growth factor receptor 3 (ErbB3)in HCC tissues and peritumoral normal tissues were detected by immunohistochemistry (IHC).The relative expressions of ErbB3 and HBx in HCC tissues and peritumoral normal tissues were detected by Western blot,and relative expressions of ErbB3 in HepG2 of which green fluorescent protein (GFP) and GFP-HBx were respectively transfected were detected.The relative expressions of ErbB3 mRNA in HepG2 transfected by GFP and GFP-HBx were detected by real-time polymerase chain reaction (RT-PCR).The migration and invasion of HepG2 were respectively detected by Transwell assay with and without matrix.The measurement data with normal distribution were represented as $± s.The comparisons between groups were evaluated with the independent-sample t test.Correlation analysis was done by the Pearson test.Results (1) The expressions of ErbB3 were detected by IHC:relative value of mean optical density (MOD) of ErbB3 in HCC tissues of 20 patients with HCC and peritumoral normal tissues of 10 patients with benign tumor of liver were 2.54± 1.33 and O.99±0.29,respectively,with a statistically significant difference (t =6.542,P ＜ 0.05).(2) The relative expressions of ErbB3 and HBx were detected by Western blot:relative expressions of ErbB3 and HBx were respectively 0.79±0.13,1.10±0.28 in HCC tissues of 10 patients with HCC and 1.07±0.17,0 in peritumoral normal tissues of 10 patients with benign tumor of liver,with statistically significant differences (t =3.229,19.486,P＜0.05).The results of Pearson test showed that there was a positive correlation of expression between ErbB3 and HBx in HCC tissues (r=O.637,P＜ 0.05).(3) The relative expressions and transcriptional levels of ErbB3 were detected by Western blot and RT-PCR:relative expressions of ErbB3 in HepG2 of which GFP and GFP-HBx were respectively transfected were O.75±0.11 and 1.10±0.10,respectively,with a statistically significant difference (t=4.291,P＜0.05).The relative expressions of ErbB3 mRNA in HepG2 of which GFP and GFP-HBx were respectively transfected were O.38±0.03 and O.94±0.07,respectively,with a statistically significant difference (t=11.703,P＜O.05).(4) The effects of ErbB3 on migration and invasion of HepG2:numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay with matrix were respectively 271± 18 and 463± 31,respectively,with a statistically significant difference (t =8.202,P＜0.05).Numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay without matrix were respectively 315±38 and 549±34,respectively,with a statistically significant difference (t =8.310,P＜0.05).Conclusion HBx protein can promote the invasion and migration of hepatocellular carcinoma cells through up-regulating expressions of ErbB3 protein.

BACKGROUND: Current studies on CD62P have focused mainly on cardiovascular diseases, while only few studies have evaluated the effects of CD62P on the development of sepsis and the association between endothelial cell injury with inflammation and coagulation. This study attended to explore the association between endothelial cell injury with inflammation and coagulation by evaluating the expression of soluble CD62P (s-CD62P) in plasma and its mechanism in patients with sepsis, thus to provide the evidence of effective treatment of sepsis with anti-adhesion therapy targeted CD62P. METHODS: A total of 70 critically ill patients with systemic inflammatory response syndrome (SIRS) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrol ed for a prospective and control study. According to the diagnostic criteria of sepsis/SIRS, the patients were divided into two groups: a sepsis group (n=38) and a SIRS group (n=32). Another 20 healthy volunteers served as a control group. Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure, diabetes and its complications. The demographics of the patients including age, sex, body mass index (BMI), smoking and alcohol addict were compared among the groups. Six mL peripheral blood samples were collected within 24-hour admission in ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P, TNF-α, and hs-CRP. And variables of coagulation function such as platelet (PLT), prothrombin (PT), activated partial thromboplastin time (APTT), D-dimer and antithrombin-III (AT-III) were analyzed during 24 hours after admission to ICU. Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated. Data were expressed as mean±standard deviation and were statistical y analyzed by using SPSS 17.0 statistical software. The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and the Kruskal-Wallis test. The relations between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson's product moment correlation coefficient analysis. Changes were considered as statistically significant if P value was less than 0.05. RESULTS: Compared with the control group and SIRS group, the sepsis group demonstrated significantly higher levels of s-CD62P, TNF-α and highly sensitive C-reactive protein (hs-CRP) (P<0.05). The plasma levels of D-dimer, PT, and APTT in the sepsis and SIRS groups were significantly higher than those in the control group, while the platelet count and the activity of AT-III were obviously lower (P<0.05). In the sepsis group, the plasma levels of hs-CRP and TNF-α were positively correlated with PT, APTT, and D-dimer, and negatively correlated with AT-III and PLT (P<0.05). The plasma levels of s-CD62P were significantly correlated with the plasma levels of TNF-α, hs-CRP, D-dimer, PT, and APTT, whereas they were correlated negatively well with PLT and AT-III (P<0.05). CONCLUSIONS: The concentration of plasma s-CD62P is elevated as a early biomarker in patients with sepsis, and it serves as one of the pathogenic factors responsible for endothelial cell damage. Coagulation and mediators of inflammation promote each other, aggravating the severity of sepsis. Plasma s-CD62P may be an important factor for the development of coagulation and inflammatory reaction.

An recombinant vector pCSV-EPO for expression of EPO cDNA in mammalian cells was constructed by the techniques of gene recombinant, PCR amplification and region-specific mutagenesis. The pCSV-EPO was introduced into COS7 cells by DEAE-dextran-mediated transfection. The expression of the EPO was demonstrated by EPO-ELISA assay. At 48h post transfection, the EPO level was 25ng/ml and 72 h was 17ng/ml.

Objective: To observe the clinical efficacy of filiform fire-needling plus continuous passive motion (CPM) therapy for frozen shoulder. Methods: A total of 72 patients were randomized into an observation group and a control group by the random number table method, with 36 cases in each group. Patients in the control group received celecoxib capsule plus CPM, while those in the observation group received filiform fire-needling plus CPM. The whole course of treatment lasted for 2 weeks in both groups. The visual analog scale (VAS), Constant-Murley score (CMS) and range of motion of shoulder joint were measured for therapeutic efficacy evaluation. Results: The total effective rate was 91.7％ in the observation group, higher than 72.2％ in the control group, and the between-group comparison showed statistical significance (P<0.05). After treatment, the VAS scores in the two groups dropped significantly, the CMS as well as the range of motion including abduction, forward flexion and extension were all increased significantly, and the intra-group comparisons showed statistical significance (all P<0.05). The betweengroup comparisons showed the improvements in these items in the observation group were more significant than those in the control group (all P<0.05). Conclusion: Filiform fire-needling plus CPM can produce more significant efficacy than celecoxib capsule plus CPM for frozen shoulder; it can alleviate pain, improve shoulder function and restore joint range of motion in such patients.

Objective To investigate the effect of diabetes on clinical efficacy of transcatheter arterial chemoembolization (TACE) in the treatment of non-viral hepatitis hepatocellular carcinoma (HCC). Methods Retrospectively analyzed the clinical data of 367 non-hepatitis virus HCC patients treated by TACE, included 153 diabetes mellitus cases (test group) and blood glucose of 214 patients was normal (control group). To assess the treatment effect after 1 month of TACE based on response evaluation criteria in solid tumors, include complete response (CR), partial response (PR), stable disease (SD), progressive disease (PD), and calculate the disease control rate. Through 6 to 75 months follow-up to observed long-term efficacy, record the time to progression (TTP) and overall survival (OS) time. Survival rate were analyzed using Kaplan-Meier method and Log-rank analysis by SPSS 16.0. The single-factor analysis was used to analyze variables which variables that differed were analyzed by Cox regression. Results The disease control rate of test group was 69.9%(107/153) and control group was 74.3%(159/214), the difference was no statistically significant (P=0.125). The median time to progression (mTTP) and median overall survival (mOS) of test group were 10.0 and 15.0 months;and the mTTP and mOS of control group were 14.0 and 19.0 months, the difference were statistically significant (P=0.023 and P= 0.026). Tumor diameter ≥4.5 cm, numbers of tumor ≥3, invasion of blood vessels, α-fetoprotein≥200 μg/L, Eastern Cooperative Oncology Group score and diabetes were risk factors for OS of HCC patients. Conclusion Diabetes is unfavorable factors for overall survival of non-hepatitis HCC tread by TACE.