Objective To investigate the differences of expression and activation of natural killer (NK) cell G2D (NKG2D) in patients with different immune status of chronic hepatitis B virus (HBV) infection, and to explore the significance of NKG2D-mediated immune injury in HBV infection.Methods Fifteen chronic HBV carriers (immune tolerance),15 chronic hepatitis B (CHB, immune activation) patients, 15 HBV-related acute/subacute-on-chronic liver failure (HBV-ACLF, immune over-activation) patients were enro1led in this study from January 2010 to December 2011 in the Third Hospital of Hebei Medical University.The frequencies of NK cells and NKG2D+ NK cells in peripheral blood mononuclear cells (PBMC) were detected by flow cytometry.The NKG2D mRNA expressions were measured by real-time fluorescent quantitative polymerase chain reaction.Localization and hemi-quantitative analysis of NKG2D+ cells in liver tissue were performed by immunohistochemistry staining.Concentrations of serum interferon(IFN)-γ, tumor necrosis factor(TNF)-α, perforin and granzyme B were quantified by enzyme 1inked immunosorbent assay (ELISA).Normally distributed continuous variables were analyzed using one-way analysis of variance (ANOVA), followed by Student-Newman-Keuls q test for evaluating variances between each two groups.For non-normally distributed data or heterogeneity of variance, differences between groups were analyzed using nonparametric Kruskal-Wallis H test, followed by Nemenyi test for pairwise comparisons.Pearson chi-square test was used to analyze categorical variables.Results The percentages of NK cells in PBMC were (13.58±3.24)% in healthy controls, (5.42±2.18)% in chronic HBV carriers, (7.92±2.85)% in HBV-ACLF group and (8.43±2.92)% in CHB group.The percentage of NK cells in PBMCs was lower in each chronic HBV-infected group compared with healthy controls (F=22.04, P<0.05).The frequency of NKG2D+ NK cells in HBV-ACLF group (18.92±5.85)% was the highest, followed by CHB group (12.85±3.39)%, healthy controls (8.45±2.86)%, and chronic HBV carriers (3.36±1.05%), with the statistically significant differences between each two groups (H=46.09, P<0.01).Intrahepatic NKG2D mRNA expression and NKG2D+ cells density were highest in HBV-ACLF group (6.58±1.86 and 30.69±6.67, respectively), followed by CHB group (3.25±0.95 and 17.36±4.13, respectively) and chronic HBV carriers (0.69±0.20 and 3.16±1.24, respectively), with the statistically significant differences between each two groups (H=52.10 and 52.73 respectively, both P<0.01).The similar patterns were observed in serum IFN-γ, TNF-α, perforin and granzyme B concentrations.Conclusions NKG2D expresses variously in patients with different immune status of chronic HBV infection.Activation of NKG2D may take part in the immune pathogenesis of chronic HBV infection.

BACKGROUND: Auditory steady-state responses (ASSR) is an objective method of hearing examination in clinic in recent years. ASSR has the frequency specificity as compared with previous auditory brainstem responses (ABR).OBJECTIVE: To investigate the accuracy of ASSR in objective hearing assessment.DESIGN: A case-control observation.SETTING: Department of Otorhinolaryngology, the First Affiliated Hospital of Sun Yat-sen University.PARTICIPANTS: The subjects in the normal hearing group were the 21 undergraduates (42 ears) were enrolled, they all had not any symptoms of ear disease, without history of noise exposure and disease of vestibule system, and they were normal in otoscopy. The outpatients and inpatients with neurosensory deafness were selected from the Department of Otorhinolaryngology, the First Affiliated Hospital of Sun Yat-sen University. All the children cases worn hearing aids, and had the speech ability, and cooperated in the examination. The main types included 6 ears of sudden deafness,8 ears of presbycusis, and 20 ears of neurosensory deafness due to other unknown causes. Central lesions were excluded by MR examination, and all the patients agreed with the enrollment. The results of pure-tone audiometry were all flat or descending audiogram. According to the severity of hearing damage, the patients were divided into mild deafness group (13ears), moderate deafness group (9 ears) and moderate-to-severe deafness group (12 ears).METHODS: ① The pure-tone audiometry was performed at the frequencies of 0.125-8 000 Hz in a sound insulation room. The auditory threshold grades of the subjects with normal hearing all accorded with the standards of GB-7583-87 expected value distribution. The average value of air-conduction auditory thresholds of pure-tone audiometry at the frequencies of 0.5, 1, 2 and 4 kHz was calculated. ② ASSR measurement was performed with the synchronous stimulation pattern in a sound and electromagnetic shielding room, including 8 points for both ears of the same stimulation intensity and the carrier frequency tones of 0.5, 1, 2 and 4 kHz respectively.③ ABR examination was performed by click sounds with sparse waves in a sound and electromagnetic shielding room, and insert earphones were used.The threshold results were judged according to the minimal stimulation sound intensity of the distinguishable Ⅴ wave. ③ The results of pure-tone audiometry were compared with those of ABR examination, and the results of ASSR measurement in different hearing groups were processed with analysis of variance, multi-classification discrimination based Bayes standard and q test.MAIN OUTCOME MEASURES: The thresholds of pure-tone audiometry, ASSR measurement and ABR examination, and the correct rate analyzed by the multi-classification discrimination based Bayes standard were mainly observed.RESULTS: The indexes of the 42 ears in the normal hearing group, 13, 9 and 12 ears in the mild, moderate and moderate-to-severe deafness groups were all involved in the analysis of results. ① The ABR values were accorded with the actual hearing levels, and the closest to the ASSR thresholds at 1-2 kHz; ASSR reflected induction rates at different frequencies were gradually decreased with the aggravation of hearing damage, and that at each frequency varied with the changes of hearing level, the induction rates of ASSR responses were all 100% for the subjects with normal hearing and patients with mild deafness, but those for the patients with moderate and moderate-to-severe deafness were decreased (0.5 kHz: 77.8%,92.8%; 4 kHz: 88.9%, 85.7%). At different frequencies, the ASSR thresholds in the moderate-to-severe deafness group were significantly higher than those in the normal hearing group (P ＜ 0.05). The ASSR thresholds at 0.5 and 4 kHz in the moderate-to-severe deafness group were significantly higher than those in the mild deafness group (P ＜ 0.05). The ASSR threshold at 2 kHz in the mild deafness group was significantly higher than that in the normal hearing group (P ＜ 0.05). The ASSR thresholds at 4 kHz in the everedeafness group were significantly higher than those in the normal hearing group and mild deafness group. ② The incorrect discriminations of actual pure-tone audiometry were analyzed with the interactive clustering discriminant analysis of ASSR measurement and actual pure-tone audiometry, and the results showed that the correct rate of discrimination was 100% in the normal hearing group; Only 1 of the 12 cases in the mild deafness group was incorrectly judged, and the correct rate was 92%; Only 1 of the 19 cases in the moderate deafness group was incorrectly judged, and the correct rate was 89%; the correct rate in the moderateto-severe deafness group was 83%.CONCLUSION: The results of ASSR measurement can detect the incorrect discrimination of objective hearing condition by taking the results of pure-tone audiometry as the standards. ASSR has an acceptable accuracy for deafness higher than mild level in estimating objective hearing, and it has a better prospect of application in practice.

@@

A lack of effective drugs and technical means to eradicate hepatitis B virus (HBV) is a bottleneck that limits the ability to fully cure HBV infection. Recently, genome-editing technology based on clustered regularly interspaced short palindromic repeats -associated protein 9 is an emerging technique for editing specific gene loci, which can specifically target HBV covalently closed circular DNA, effectively inhibits HBV DNA replication and regulates HBV functional protein expression, and is expected to become a powerful gene therapy tool for the complete eradication of HBV. Considering this, it has become the focus of attention for scholars at home and abroad that how to use clustered regularly interspaced short palindromic repeats -associated protein 9 to accomplish modification of HBV genomes for complete eradication of HBV. This paper summarizes the latest progress based on the latest research results at home and abroad in the application of clustered regularly interspaced short palindromic repeats -associated protein 9 gene editing technology in anti-HBV infection treatment, and expounds its potential and challenges as a radical cure for HBV infection.

Objective: To investigate the knowledge of nonalcoholic fatty liver disease (NAFLD) among medical staff and general population in Hebei Province, China, understand the degree of awareness of NAFLD among residents, standardize the diagnosis and treatment of NAFLD, improve the correct awareness of NAFLD among general population, promote the establishment of a reasonable lifestyle, and scientifically and effectively control the incidence and mortality rates of NAFLD. Methods: The investigation was performed in 30 hospitals and neighboring schools and communities in Hebei Province, and a questionnaire survey on the knowledge of NAFLD was performed for 1 300 medical workers and 2 000 persons from the general population. EpiData3.1 software was used to input data, and SPSS21.0 was used for statistical analysis. The chi-square test was used for comparison of rates between groups. Results: Of all medical staff, 39.0% knew that liver biopsy was the gold standard for the diagnosis of NAFLD; 63.7% thought liver biopsy had the highest diagnostic value, followed by liver ultrasound (61.0%) and liver CT (48.1%); only 1.2% thought improvement of insulin resistance was the preferred treatment for NAFLD. Among 486 medical workers who had participated in the diagnosis and treatment of NAFLD, only 1.2% thought that the patients had good compliance. Of all persons from the general population, 15.2% thought NAFLD was infectious and 58.6% thought NAFLD might progress to liver cirrhosis or liver cancer. Of all respondents with body mass index ≥28 kg/m², 17.8% did not know that he/she had obesity, and as high as 55.8% were obese or did not control body weight within the past 1 year. Of all NAFLD patients, 50.5% were complicated by hyperlipidemia, 18.5% were complicated by diabetes, and 29.5% were complicated by hypertension; 43.0% had not received standardized treatment, and among the other 57.0% respondents who received treatment, 79.0% were given non-drug treatment such as reasonable diet and physical exercise. In NAFLD patients who were not improved after treatment, the major cause of failure was that they were not able to stick to healthy living habits. Most NAFLD patients were willing to obtain the knowledge of disease through television, website, and newspaper. Conclusion Medical staff do not have enough awareness of NAFLD, and some of them cannot perform standardized diagnosis and treatment of NAFLD with reference to clinical guidelines. Therefore, the education on NAFLD should be enhanced for medical staff to guide them to perform accurate screening, diagnosis, evaluation, and treatment of NAFLD. The general population has low awareness of NAFLD and NAFLD patients have poor treatment compliance, which are important causes of increased incidence rate of NAFLD and disease progression. The general population should be educated about NAFLD to help them establish a reasonable lifestyle.

Recent years found a series of severe incidents of injuring or killing medical workers in several places in China,deteriorating patient-physician relations,and disturbing medical orders.Beijing Administration of Hospitals,since its founding,has attached great importance to the management of medical disputes,as evidenced in the full-process management covering pre-during-post disputes.The administration guided the hospitals in their dispute resolution,and included resolution rate of disputes as an index of municipal hospital performance evaluation.By means of the guidance of such evaluation, medical disputes and major medical accidents have been resolved satisfactorily,and fine management of service quality of such hospitals in terms of medical service,pharmacy,nursing and medical technology has been intensified all the time.

Phage antibody display technology is currently the most widely used in vitro antibody screening technology,which uses bacteriophages as a vector,and inserts exogenous antibody library genes into phage capsid protein genes,and expresses the capsid protein on the phage surface while also displays the antibody protein.Antibody drugs play an important role in tumor immunity and microbial immunity due to their targeting advantages,which is also an important driving force for them to become a hot spot in the field of pharmaceutical research and development.Therefore,this article reviews the background,basic principles,antibody library types and antibody fragment types of phage display technology,and looks forward to the latest progress and application prospects of fully human antibodies.

With the rapid development of network technology and the situation of COVID-19 pandemic, the way people use the Internet has changed dramatically, causing the original network behavior to change again and again, and with its huge impact on people's mental and physical health.This paper deeply elaborate the connotation and development of network behavior and analyzes the impact of network behavior on people's health under COVID-19, then puts forward suggestions to speed up the construction of information infrastructure, strengthen network legislation, improve the information literacy of the whole population, and purify the network environment.

Objective To evaluate the clinical efficacy of bronchial artery embolization in the treatment of massive hemoptysis ,and to evaluate the clinical efficacy and adverse reactions of 3-year recurrence and survival . Methods 80 patients with hemoptysis or chronic recurrent hemoptysis were selected as the research subjects , and they were randomly divided into two gourps according to the digital table ,40cases in each group.The interventional embolization group was given bronchial artery embolization .The drug group was treated with phentolamine and pituitrin.The index of the system were observed and evaluated:(1)the effect of controlling massive hemoptysis;(2) the adverse reaction of the patients;(3) the recurrence rate of 3 years.Results (1) The total effective rate of the interventional embolization group was 92.5％,which was significantly higher than 77.5％ of the drug group (χ2 =9.044,P<0.05);(2) The incidence rate of adverse reaction of the interventional embolization group was 12.5％, which was lower than 35.0％of the drug group ,the difference was not statistically significant between the two groups (χ2 =1.742,P>0.05).(3) The 3 years recurrence rate of the interventional embolization group was 7.5％,which of the drug group was 40.0％,the difference between the two groups was statistically significant (χ2 =12.557,P<0.05).The 3 years survival rate of the interventional embolization group was 95.0％,which of the drug group was 92.5％,the difference between the two groups was not statistically significant (χ2 =0.215,P>0.05).Conclusion The bronchial artery embolization in the treatment of hemoptysis has ideal clinical effect .It is a minimally invasive and effective clinical treatment method ,and the effect has safety ,less adverse reaction and low recurrence rate ,which is worthy of extensive promotion and application .

Objective:To construct a recombinant plasmid pET30a-leucine-rich repeat (LRR) containing 15 (LRRC15) of Taenia solium, prokaryotically express and purify the LRRC15 recombinant protein, and prepare a rabbit polyclonal antibody. Methods:The LRRC15 protein encoding gene of Taenia solium was obtained by whole gene synthesis; it was cloned into pET30a vector, and the recombinant plasmid pET30a-LRRC15 was constructed and identified by double-enzyme PCR; the recombinant plasmid was transformed into competent cells of Escherichia coli BL21 (DE3), and the recombinant protein LRRC15 was induced to express by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression product was analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); the LRRC15 recombinant protein was purified by Ni-IDA affinity columns, the purified recombinant protein was analyzed and identified by SDS-PAGE, and the specificity of the purified recombinant protein was identified by Western blot (WB); the New Zealand rabbits were immunized with purified LRRC15 recombinant protein to prepare polyclonal antibodies against LRRC15, and the potency of the purified polyclonal antibody was determined by indirect enzyme-linked immunosorbent assay (ELISA). Results:After PCR identification, a band with a length of 1 506 bp was amplified, which was consistent with the LRRC15 gene; after SDS-PAGE and WB identification, the LRRC15 target protein with a relative molecular mass ( Mr) of about 55.36 × 10 3 was obtained; after immunizing New Zealand rabbits with purified LRRC15 recombinant protein, a polyclonal antibody against LRRC15 was obtained, and its potency was 1∶1 587 200. Conclusion:The recombinant plasmid pET30a-LRRC15 is successfully constructed, the LRRC15 recombinant protein of Taenia solium is prepared, and a high purity and high potency rabbit anti polyclonal antibody against LRRC15 recombinant protein is obtained.