AIM:To clarify the impact of heart shock factor 4b (Hsf4b) K65R mutation on the regulation of downstream protein expression .METHODS:Non-functional Lys mutant plasmid pWZL-blast-HA-Hsf4b/K65R was genera-ted by replacing single , homologous amino acids using KOD-Plus-Mutagenesis-Kit.Mouse lens epithelial mLEC stable cell lines expressing Hsf4b or Hsf4b/K65R were constructed by lentivirus infection .The expression of Hsf4b in the mutant and the wildtype mLEC cells was confirmed by Western blotting .The expression of Hsf4b downstream proteins such as heat shock protein ( Hsp)70, Hsp90, Hsp27 and CryAB was examined by Western blotting and real-time PCR.RESULTS:The results of PCR and DNA sequencing confirmed the successful construction of mLEC Hsf 4b/K65R mutant.The K65R mutation didn’t influence Hsf4b expression in the mLEC cells.After K65R mutation in Hsf4b, the expression levels of Hsp27 and CryAB were down-regulated and the expression of Hsp 70i and Hsp90a upregulated.CONCLUSION: pWZL-blast-HA-Hsf4b/K65R can be used to construct a stable cell line by infecting with lentivirus .Hsf4b/K65R mutation influ-ences the regulation of downstream heat shock proteins .

In view of the current situation of the teaching of diagnostics, this paper analyzes the ex-isting problems and deficiencies , and puts forward the teaching reform of the National Medical College Students' clinical skills competition. Through the construction of experimental teaching demonstration center of clinical skills, we set up theclinical skills associationamong the students, boldly try innovative training, strengthen humanistic education and take other measures to improve the quality of clinical practice teaching of diagnostics, so as to achieve the purpose of the competition focusing on promote teaching, learning and using by competition.

Objective To definite the status of tone of autonomic nervous system during water immersion process by analyzing the nonlinear characteristic of ECG signals;To explore the relation of approximate entropy(ApEn),scaling exponent(?) changes and modulation function status of autonomic nervous system.Methods The ECG signals of rats during the process of restraint stress(RS),water-immersion restraint stress(WRS) and Vagotomy and Water-immersion restraint stress(vagotomy-WRS) were recorded.The ApEn and ? of RR intervals were calculated.Results After the stress,the differences of the value of ApEn and ? of RR intervals between RS and vagotomy-WRS groups and between different periods in vagotomy-WRS group were not significant.The value of ApEn increased in WRS group and decreased in RS group significantly.Compared with pre-stress,? showed decreased trend,all of the values smaller than 0.5 in WRS group and increased obviously,most values larger than 0.5 in RS group.Conclusion The increased ApEn and decreased ? indicates a higher intensity of parasympathetic activity.The parasympathetic activity is the predominant response of autonomic nervous system during water immersion process.

Objective To investigate whether the percutaneous ethanol injection (PEI)under sedation and analgesia can in-crease the energy deposition and curative efficiency of the high intensity focused ultrsound(HIFU)in treating unresectable middle and advanced stages of primary liver cancer.Methods Thirty-six cases of clinically diagnosed unresectable middle and advanced sta-ges of primary liver cancer were randomly divided into the PEI+ HIFU group(combination group,n = 23)and the simple HIFU group (HIFU group,n=13);10mL of the mixture of 99.7% ethanol and iodized oil (9:1)was given by intratumoral injection at 30 min before ablation in the PEI+HIFU group,while 0.9% physiological saline 10mL was replaced in the simple HIFU group.The ablation energy efficiency factor(EEF)and irradiation time were compared between the two groups.Results The ablation EEF in the PEI+HIFU group and the simple HIFU group were (13.82+4.26)J/mm3 and (25.63+6.31)J/mm3 respectively,the PEI+HIFU group was significantly lower than the simple HIFU group (P <0.05);the irradiation time were (1 468.28+253.21)s and (2 352.56+463.34)s respectively;which in the PEI+ HIFU group was significantly shortened (P <0.05).Conclusion PEI can enhance the HIFU ablation energy deposition and improve the efficiency of HIFU for treating unresectable primary liver cancer.

Objective:To establish the HPLC fingerprint of Centellae herba and determine the content of asiaticoside, madecassic acid and asiaticoside B simultaneously; To compare the quality differences of Centellae herba collected in different months. Methods:The chromatographic condition was a Shimadzu InertSustain C18 column (4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile and 2 mmol/L beta cyclodextrin in gradient elution at the flow rate of 0.8 ml/min. The detection wavelength was 204 nm, and the column temperature was 30 ℃. The different Centellae herba materials of collected in 2-12 months from Chenzhou were studied by the similarity evaluation combined with cluster analysis, principal component analysis and the three contents determination. Results:The HPLC fingerprint of Centellae herba was established and 9 common peaks were designated. The eleven samples were different, which can be aggregated into 4 categories and the quality of Centellae herba collected in July was the best. Conclusion:The established fingerprint and multi-components quantitative method are stable and reliable, which can provide a reference for the quality control and the utilization of Centellae herba resource.

Nowadays, more attentions have been paid on brown adipose tissue (BAT) because BAT plays a great role in individuals with obesity and metabolic diseases. 18F-fluorodeoxyglucose (FDG) PET/CT is noninvasive and sensitive to detect the location, volume, and metabolic activity of BAT. 18F-FDG PET/CT for BAT provides new insight for the prevention and clinical management of obesity and diabetes. The review summarizes the methods to activate BAT, the preparation of patients, 18F-FDG PET/CT imaging techniques and clinical application.

Objective:To explore values and differences of multi-modality image registration and normalization methods in 18F-AV45 PET quantitative analysis of Alzheimer′s disease (AD). Methods:Twenty AD patients (10 males, 10 females; age (77.0±5.8) years) and 20 normal controls (NC; 8 males, 12 females; age (75.2±4.8) years) from the AD neuroimaging initiative (ADNI) open database of the National Institute on Aging were analyzed. β-amyloid (Aβ) deposition (positive/negative) was assessed by visual analysis. The SUV ratio (SUVr) in each brain region and individual average SUVr were calculated using template normalization method (M1), normalization after registration with 18F-FDG PET or MRI image (M2 or M3) respectively with the cerebellum as the reference area. The intra-class correlation coefficient (ICC) was used to complete the reliability between methods, and independent-sample t test and one-way repeated measures analysis of variance were used to compare the differences of quantitative indexes between different groups and different methods. ROC curve analysis was used to compare the diagnostic efficacy in distinguishing AD and NC, Aβ positive and negative cases. Results:There were 15 and 6 patients with positive Aβ deposition in AD group and in NC group respectively by visual analysis. The SUVrs of three methods were with good consistency (ICC=0.82, P<0.001), and the differences among individual average SUVrs (1.29±0.17, 1.36±0.23, 1.45±0.24) were significant ( F=68.78, P<0.001). There were significant differences between AD group (1.39±0.17, 1.48±0.24, 1.58±0.25) and NC group (1.20±0.10, 1.24±0.15, 1.33±0.16; t values: 3.55-4.33, all P<0.05), Aβ positive group (1.39±0.16, 1.50±0.21, 1.59±0.23) and negative group (1.19±0.11, 1.21±0.14, 1.31±0.15; t values: 4.58, 5.11, 4.41, all P<0.001), and the individual average SUVr of M3 was higher (both P<0.001). The AUCs of distinguishing Aβ positive and negative deposition of M1-M3 were 0.86, 0.88, 0.84 and the thresholds of SUVrs were 1.29, 1.37, 1.52, respectively. Conclusion:The three multi-modality registration and normalization methods are reliable methods for quantitation of 18F-AV45 PET imaging with certain differences, and should be selected carefully based on data conditions in practice.

An ECG information database system was established with 1297 ECG information records and clinical informations. The data and the analysis software in this database can be shared with the international physiological signal databases such as MIT-BIH Arrhythmia. The establishment of this database will facilitate the clinical interpretation, diagnosis and prediction of cardiovascular diseases, especially the cardiac sudden death (SCD).
Artificial Intelligence ; Database Management Systems ; Databases, Factual ; Electrocardiography ; statistics & numerical data ; Heart Diseases ; diagnosis ; Humans

In recent years, the potential value of nonstructural protein (NSP) 2C was well documented for distinguishing foot-and-mouth disease virus (FMDV) in infected animals and vaccinated animals. In order to develop a more sensitive approach to detect natural infected FMDV while there is no interact with vaccinated FMDV, we incorporated a major epitope region of 2C with whole 3AB coding region within NSP and expressed in Escherichia coli. We got a 47.6 kD fusion protein named 2C'3AB. The product showed a specific reactivity with FMDV from serum of infected animal by using Western blotting analysis. This suggests that this protein could be applied to distinguish infected FMDV and vaccinated FMDV. We further compared 2C'3AB protein with 3ABC fusion protein, another available protein used for detecting infected FMDV, using indirect ELISA assay. The results showed that 2C'3AB-ELISA had higher sensitivity than that of 3ABC-ELISA for distinguishing infected FMDV and vaccinated FMDV of sera from epidemic region. Therefore, this recombinant protein 2C'3AB is a good candidate protein to develop more sensitive method to differentiate infected FMDV and vaccinated FMDV from vaccinated animals. This finding will increase our capability to check the infectious virus carrier and finally improve FMDV infection control.
Animals ; Antibody Specificity ; Carrier Proteins ; genetics ; immunology ; metabolism ; Epitopes ; immunology ; Escherichia coli ; genetics ; metabolism ; Foot-and-Mouth Disease Virus ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Viral Nonstructural Proteins ; genetics ; immunology ; metabolism

Objective: To retrospectively analyze the distribution of non-fermentative bacteria causing bloodstream infection in hospitalized patients in Sichuan Province and their drug susceptibility to common antibiotics for better understanding their epidemiological characteristics. Methods: From January 1, 2015 to December 31, 2017, all of the non-fermentative bacteria isolated from patients with bloodstream infection in nine hospitals in Sichuan Province were collected. Species distribution and drug resistance test results were retrospectively analyzed. Results: A total of 6 291 strains of pathogenic bacteria were isolated, including 3 674 strains of gram-negative bacteria (58.4%) and 2 617 strains of gram-positive bacteria (41.6%). The gram-positive bacteria were 1 895 strains of Staphylococcus (30.1%), 372 strains of Streptococcus (5.9%), 317 strains of Enterococcus (5.1%) and 33 strains of other gram-positive bacteria (0.5%). The gram-negative bacteria were 3 191 Enterobacteriaceae strains (50.7%), 389 non-fermentative strains (6.2%) and 94 other gram-negative strains (1.5%). The isolated non-fermentative bacteria were mainly Pseudomonas aeruginosa (136 strains, 35.0%), Acinetobacter baumannii (126 strains, 32.0%) and Stenotrophomonas maltophilia (33 strains, 8.5%). There were 167 (42.9%), 112 (28.8%) and 82 (21.1%) non-fermentative bacteria isolated in internal medicine departments, ICUs and surgery departments, respectively. The drug resistance rates of Pseudomonas aeruginosa to cefepime, ciprofloxacin and gentamicin were 16.5%, 10.7% and 9.9%, respectively. Carbapenems-resistant Pseudomonas aeruginosa (CR-PA) accounted for 13.6%. No polymyxin-resistant Pseudomonas aeruginosa was found. The resistance rates of Acinetobacter baumannii to all antibiotics were over 30% except for minocycline and polymyxin and 75.7% of carbapenem-resistant Acinetobacter baumannii (CR-AB) were isolated. The drug resistance rates to levofloxacin, trimethoprim/sulfamethoxazole and ceftazidime were 0%, 0% and 37.0% in Stenotrophomonas maltophilia isolates and 10.5%, 4.2% and 19% in Burkholderia cepacia isolates, respectively. Conclusions Non-fermentative bacteria causing 6.2% of bloodstream infection in Sichuan, mainly by Pseudomonas aeruginosa and Acinetobacter baumannii. There were significant differences in the isolation rate of non-fermentative bacteria in different departments and most of the strains were isolated in internal medicine departments. The detection rate of Acinetobacter baumannii with multiple drug resistance was high, while other non-fermentative bacteria had good sensitivity to antibiotics.