Objective To investigate the efficacy of entecavir (ETV) versus tenofovir disoproxil fumarate (TDF) and the treatment measures for poor response in previously untreated chronic hepatitis B (CHB) patients with high viral load. Methods A total of 165 CHB patients who received antiviral therapy and met the inclusion criteria in Department of Infectious Diseases, The First Affiliated Hospital of Guangxi Medical University, from June 2016 to July 2021 were enrolled. The patients enrolled had a baseline HBV DNA level of > 6lg copies/ml and were previously untreated CHB patients who had used ETV or TDF for 48 weeks, and quantitative real-time PCR was used to measure HBV DNA. Virologic response rate was calculated after 48 weeks of treatment; a logistic regression analysis was used to investigate the influencing factors for the response of HBV DNA < 500 copies/mL and HBV DNA < 100 copies /mL at 48 weeks; a stratified analysis was performed to compare the virologic response rate of HBV DNA < 500 copies /ml and HBV DNA < 100 copies/ml after 48 weeks between the patients with different ages, sexes, baseline HBV DNA levels, baseline alanine aminotransferase (ALT) levels, types of first-line medication, and HBeAg statuses. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups, and the binary logistic regression model was used for multivariate analysis. Results After 48 weeks of treatment, 85.5% (141/165) of the patients achieved an HBV DNA load of < 500 copies/mL, and 66.1% (109/165) of the patients achieved an HBV DNA load of < 100 copies /mL, with no significant difference in treatment outcome between the ETV group and the TDF group. The multivariate logistic regression analysis showed that sex( OR =2.793, 95% CI : 1.197-6.517), baseline HBV DNA( OR =0.369, 95% CI : 0.142-0.959), baseline ALT( OR =4.556, 95% CI : 1.770-11.732), and baseline HBeAg( OR =0.120, 95% CI : 0.033-0.429) were influencing factors for complete virologic response(all P < 0.05). For the patients with normal ALT (≤40 U/L) at baseline, 75.6% (34/45) achieved an HBV DNA load of < 500 copies/mL after 48 weeks of treatment, and 53.3% (24/45) achieved an HBV DNA load of < 100 copies/mL, with no significant difference in treatment outcome between the ETV group and the TDF group. For the patients with abnormal ALT (> 40 U/L) at baseline, 89.2% (107/120) achieved an HBV DNA load of < 500 copies/mL after 48 weeks of treatment, and the proportion of such patients in the TDF group was significantly higher than that in the ETV group (96.1% vs 84.1%, χ 2 =4.386, P =0.036); 70.8% (85/120) achieved an HBV DNA load of < 100 copies/mL, the proportion of such patients was no significant difference between the TDF group and the ETV group (78.4% vs 65.2%). The response of HBV DNA < 100 copies/ml of the normal baseline ALT group and the abnormal baseline ALT group, there were no significant differences between the patients aged≤30 years and aged > 30 years (77.8% vs 47.2%, 85.2% vs 66.7%). For the patients who did not achieve complete virologic response (HBV DNA ≥100 copies/mL) after 48 weeks of treatment, 87.9% (29/33) achieved complete virologic response after the original treatment regimen was prolonged for 48 weeks, and 100% (9/9) of the patients achieved complete virologic response after switching to or adding the first-line nucleos(t)ide analogues (NUCs) without cross-resistance sites with the original regimen for another 48 weeks. Conclusion The patients aged > 30 years should receive antiviral therapy as early as possible, regardless of viral load and ALT level, especially those with a family history of liver cirrhosis or hepatocellular carcinoma; the patients aged ≤30 years who have a normal ALT level and a high viral load should consider initiating antiviral therapy after providing informed consent. For the patients with poor response after 48 weeks of treatment, first-line NUCs without cross-resistance sites with the original regimen should be switched to or added in time.

Objective:To investigate drug resistance gene in Mycoplasma pneumoniae(MP) and the distribution of 13 respiratory pathogens in bronchoalveolar lavage fluid(BALF) of children with Mycoplasma pneumoniae pneumonia(MPP).Methods:A total of 100 BALF of children with MPP in Peking University Third Hospital and Peking University First Hospital from January 2018 to January 2019 were collected.Fluorogenic quantitative PCR was used to detect nucleic acid and it′s drug resistance gene of MP and multiple PCR method was adopted to detect influenza A virus, influenza A virus-H 1N 1, influenza A virus-H 3N 2, influenza B, human parainfluenza virus, adenovirus, human bocavirus, human rhinovirus, Chlamydia pneumoniae, human metapneumovirus, MP, human coronavirus, and respi-ratory syncytial virus gene, and the results were compared by using Chi square test. Results:In 100 BALF samples, MP and drug resistance gene were detected by fluorogenic quantitative PCR.Totally, 83 cases (83.00%) were MP positive and 78 cases (93.98%) were drug resistant.All of them had the point mutations A2063G in V region of 23S rRNA domain.A total of 13 kinds of respiratory pathogens were detected by multiplex PCR method, and 89 cases (89.00%) were positive.Totally, 79 cases (79.00%) were MP positive, of which 74 cases (74.00%) detected only MP, and 5 cases (5.00%) detected MP combined with other pathogens.Other pathogens were detected in 10 cases (10.00%). The virus detection rate of 0-4 years old group was higher than that of >4-6 years old group ( P=0.042) and >6 years old group ( P=0.002), and the differences were statistically significant. Conclusions:MP can be detected in most BALF samples of MPP children, the drug resistance phenomenon is serious, and the main point mutation is A2063G.There were other respiratory pathogens and 2 or 3 pathogens were detected in a small number of BALF samples.

Objective:To investigate the common bacteria in the oropharynx of children with Mycoplasma pneumoniae pneumonia (MPP) and its clinical significance.Methods:A total of 134 children with MPP who were hospitalized in the Department of Pediatric Respiratory, Shengjing Hospital of China Medical University from December 2016 to June 2017 were selected as the research subjects, and 42 healthy children in the same hospital were selected retrospectively as the healthy control group during the same period.Fluorescent quantitative polymerase chain reaction Taqman probe was used to detect common oropharyngeal bacteria[ Streptococcus pneumoniae(SP), Moraxella catarrhalis(CTA), Haemophilus influenza(HI)] for the enrolled children.Firstly, the bacterial detection rate of MPP children and healthy children was compared.Then, according to age(<1 years old, 1-<3 years old, 3-<6 years old and 6-14 years old), bacterial detection[Mycoplasma pneumoniae(MP), MP+ bacteria]and bacterial species(MP+ SP, MP+ CTA, MP+ HI), 134 children with MPP were divided into groups to compare.Moreover, the relevant clinical datas were retrospectively analyzed by rank sum test and chi- square test. Results:Among 134 children with MPP, 79 (58.96%) children were detected bacteria, and 17 (40.48%) children were detected bacteria among 42 healthy children, with statistically significant differences( χ2=4.404, P<0.05). Compared with the MP group, the level of white blood cell (WBC)[8.5(6.7, 12.0)×10 9/L vs.7.8(5.8, 9.3)×10 9/L, Z=-2.232], C reactive protein(CRP)[19.2(7.2, 35.0) mg/L vs.8.4(3.4, 24.6) mg/L, Z=-2.810], lactate dehydrogenase(LDH)[286(244, 365) U/L vs.250(210, 302) U/L, Z=-2.474] and the incidence of lobar pneumonia[40.51%(32/79 cases) vs.18.18%(10/55 cases), χ2=7.510], pleural effusion[13.92%(11/79 cases) vs.3.64%(2/55 cases), χ2=3.917], refractory Mycoplasma pneumoniae pneumonia (RMPP)[34.18%(27/79 cases) vs.18.18%(10/55 cases), χ2=4.151] in MP+ bacteria group were higher; the course of fever[10(7, 12) d vs.8(6, 10) d, Z=-2.706] and duration of antibiotic use[16(13, 19) d vs.12(9, 16) d, Z=-3.747] in MP+ bacteria group were longer (all P<0.05). The level of WBC in MP+ SP group[12.20(7.80, 17.30)×10 9/L] was higher than that in MP+ HI group [6.75(5.37, 9.44)×10 9/L], and the differences were statistically significant( Z=11.574, P<0.05), and the incidence of lobar pneumonia in MP+ SP group [56.67%(17/30 cases)]was higher than that in MP+ CTA group [0(0/3 cases)]and MP+ HI group[18.75%(3/16 cases)], and the differences were statistically significant( χ2=9.770, P<0.05). Conclusions:Bacterial colonization or infection is more likely to occur in the oropharynx of children with MPP.When WBC, CRP, and LDH are significantly increased and the image shows a large consolidation or pleural effusion, it may indicate mixed bacterial infection, longer course of fever and higher incidence of RMPP, and the common mixed bacteria is SP.

Objective:To understand the pathogen distribution of children with influenza in North China in the past 2018-2019 years, and compare the accuracy of influenza virus antigen test results with that of influenza virus nucleic acid test results, provide reference data for clinical use good influenza virus pathogen detection methods.Methods:Five hundred and eighty throat swab samples of influenza-like children in 10 hospitals, northern China, were collected from December 2018 to January 2019.Each sample was tested by rapid influenza diagnostic test and reverse-transcription polymerase chain reaction(RT-PCR).Results:Of all 580 clinical samples, 256 positive samples (256/580 cases, 44.14%)were detected by the influenza rapid influenza diagnostic test, of which 235 were pure influenza A(235/256 cases, 91.8%), 21 cases were pave influenza B(21/256 cases, 8.2%), and 324 case were negative samples(324/580 cases, 55.86%). No cases were detected positive A and B at the same time.Of all 580 samples were detected using the A /B influenza virus RT-PCR, and a total of 353 cases(353/580 cases, 60.9%) were positive (of which 242 cases were influenza virus antigen-positive), of which 311 were pure A influenza(311/353 cases, 88.1%) and 41 were pure B influenza(41/353 cases, 11.6%), 1 case of mixed infection of A and B(1/353 cases, 0.3%), and 227 cases were negative(227/580 cases, 39.1%). In 324 cases of influenza virus antigen negative samples, 111 cases(111/324 cases, 34.3%) were positive for influenza virus nucleic acid.The detection rate of influenza A in Taiyuan was 23.2% (22/95 cases), and the detection rate of influenza B was 43.2% (41/95 cases), which was significantly different from other regions.With reverse-transcription polymerase chain reaction detection as the standard, the diagnostic value of influenza pathogen detection reagents was evaluated.The sensitivity, specificity, missed diagnosis rate, misdiagnosis rate, positive predictive value, negative predictive value, positive likelihood ratio, negative likelihood ratio, Youden index and area under the receiver operating characteristic curve were 68.56%, 93.83%, 31.44%, 6.17%, 94.53%, 65.74%, 11.12, 0.335, 0.624 and 0.812.Conclusions:From December 2018 to January 2019, the majority of children′s influenza in northern China is influenza A virus.Except Taiyuan which is dominated by influenza B. Influenza virus nucleic acid detection has high sensitivity and specificity for diagnosing influenza, and also has the ability to distinguish virus subtypes.Influenza virus antigen detection has a certain diagnostic value, a good specificity (93.83%), sensitivity (68.56%) which needs to be further improved, and a certain rate of missed diagnosis (31.44%) needs to be paid attention to possible missed diagnosis.Detecting positive cases of influenza virus antigens should be given a fast and effective anti-viral treatment, while the negative cases, especially those at high risk for influenza complications, should be confirmed influenza virus RT-PCR as soon as practical.

Objective:To retrospectively analyze the serological, virological, biochemical, liver histological status and clinical outcomes in HBeAg-negative chronic hepatitis B (CHB) patients with low HBV viral load, and to explore the necessity of antiviral therapy for these patients.Methods:A total of 99 HBeAg-negative CHB patients with HBV DNA level < 4 lg copies/ml who performed liver biopsy at the baseline were enrolled from the follow-up cohort. Among them, 23 cases received the second liver biopsy during follow-up. The relationships among the degree of inflammation and fibrosis of liver tissues, the status of HBsAg and HBcAg, age, gender, family history, HBV DNA load, serological markers and other indicators were analyzed. The pathological differences between two liver biopsy examinations were compared. The effect of nucleos(t)ide analogues (NAs) treatment on patient’s clinical outcomes were analyzed. For multivariate analysis, a binary logistic regression model was performed. Log-rank test was used to compare the cumulative incidence of hepatocellular carcinoma (HCC) in NAs-treated and non-NA streated patients.Results:Baseline liver histology status showed that 58.6% (58/99) patients had obvious liver tissue damage in their baseline liver tissue pathology (G≥2 and /or S≥2). Univariate logistic regression analysis showed that a liver cirrhosis (LC) family history, a HBsAg-positive family history, baseline alanine aminotransferase and aspartate aminotransferase levels were positively correlated factors for liver tissue damage. Multivariate logistic regression analysis showed that a LC family history was the main risk factor for liver tissue damage. Twenty-three cases had received a second liver biopsy after an interval of 4.5 years. In 10 untreated cases, the second liver biopsy results showed the rate of obvious liver tissue damage (G≥2 and/ or S≥2) increased from 50.0% to 90.0%. In the other 13 cases who received NAs treatment, the second liver biopsy showed improvement in liver histology, and the rate of obvious liver tissue damage decreased from 61.5% to 46.2%. The 5-year HCC cumulative incidence in non-NAs-treated patients was significantly higher than that of in NAs-treated patients (17.7% vs. 3.8%, P = 0.046). Conclusion:For most HBeAg-negative CHB patients with low viral load, liver tissue pathology result suggests that it meets the indications for antiviral therapy, especially in patients with a LC familial history. Without antiviral therapy, liver tissue damage for these patients will progressively worse with the high incidence of HCC. Therefore, it is suggested that antiviral therapy should be started as soon as possible for the HBeAg-negative CHB patients with low viral load regardless of the alanine aminotransferase level, especially in patients over 30 years-old with a LC or HCC family history.

Objective To investigate the antibacterial effect of Fusidic acid on Mycoplasma pneumoniae and antibiotic resistant Mycoplasma pneumoniae in vitro.Methods Twenty-eight clinical strains of Mycoplasma pneumoniae isolated from patients with respiratory tract infection at Beijing Friendship Hospital Affiliated to the Capital University of Medical Sciences from January to December 2016 and 2 Mycoplasma pneumoniae reference strains were enrolled.The minimum inhibitory concentration (MIC) of Fusidic acid and Azithromycin were determined by using micro-dilution ration method.The chessboard method was used to check the antibacterial effect of combination between Fusidic acid and Azithromycin.The antibacterial activity of the Fusidic acid was evaluated by measuring the antibacterial rate of different concentrations.Results One isolate showed no mutation in 23SrRNA,26 isolates had one point mutation in loci 2063 and 1 isolate had one point mutation in loci 2064 among the 28 clinical isolates.The findings by micro-dilution method results showed that the MIC values of all the clinical isolates with mutations associated with macrolide resistance to Azithromycin were ＞ 1.000 0 mg/L,and the MIC values of all the clinical isolates with no mutations to azithromycin were ＜ 0.500 0 mg/L.The findings by micro-dilution method results showed that the MIC value of Fusidic acid for Mycoplasma pneumonia and drug resistance Mycoplasma pneumoniae was 1.000 0 mg/L.The Fractional Inhibitory Concentration index of Fusidic acid and Azithromycin combination was ≤0.500 0 mg/L.When the concentration of the Fusidic acid was lower than or equal to 32 MIC,the antibacterial effect of Fusidic acid against Mycoplasma pneumoniae increased with its higher concentration.When the concentration of the Fusidic acid was lower than or equal to 8 MIC,the longer the strain was exposed to the drug,the stronger antibacterial effect was against Mycoplasma pneumoniae.Conclusion If the treatment of Mycoplasma pneumoniae infection is not effective or the infection of patient is combined with bacteria,the application or combination of Fusidic acid may inhibit pathogenic bacteria effectively.Of course,how to use Fusidic acid in clinical treatment needs further study and discussion.

Objective To developed A laboratory diagnosis of Moraxella catarrhalis by an laboratories diagnostic method real-time fluorescence quantitative PCR assay. Methods The specific primers and probes were designed based on the sequence of outer membrane protein CopB(copB)gene in Moraxella catarrhalis,and the Taqman probe RT-PCR method was developed to detect the Moraxella catarrhalis.The standard plasmids ex-tracted from the Moraxella catarrhalis standard strains were used to constitute the standard samples,and compared with these standard samples,the sensitivity of the fluorescence quantitative PCR assay was tested by the estab-lished standard curves.The specificity of the fluorescence quantitative PCR assay was tested by the DNA samples of other bacterias in the laboratory.Meanwhile,321 throat swab samples from inpatient and outpatient child pa-tients,with asthma infection were collected as clinical samples to validate the fluorescence quantitative PCR as-say.Results The standard curve was drawn in the real-time PCR by the Taqman fluorescence reporter.During the sensitivity tests,the newly-developed real-time fluorescence PCR could detect at least 10 copies of Moraxella catarrhalis,and could successfully distinguish several DNAs of the pathogens.On the basis of the validation result of the 321 throat swab samples,there are 25 Moraxella catarrhalis with 7.79 % positive rate.Conclusion The fluorescence quantitative PCR assay is of great sensitivity and specificity,and it can be widely used for the detec-tion of Moraxella catarrhalis.

Objective To develope a new Real-time quantitative PCR assay using SYBR green as fluorescence reporter, which is rapid, specific, sensitive, cheap and accurate for the detection of mycoplasma pneumoniae(MP), and evaluated its clinical application value.Methods The sequence of the 23S rRNA gene in MP type strain FH was selected as amplified regions, and specific primers were designed.Then the related plasmids were extracted as standards,and the absolute quantitative standard curve was established.The sensitivity ,specificity of the fluorescence quantitative PCR assay was compared with the nest-PCR and kit;To calculate correlation coefficient, coincidence rate and kappa coefficient, clinical samples were detected using above-mentioned methods and cultivation,respectively.Results The detection sensitivity of the new real-time PCR and nest-PCR was 10 copies of FH DNA,while the kit 100 copies.In the specificity tests,the MP sample was positive,while mycoplasma hominis and other four bacteria were all negative.We applied this real-time PCR assay ,nest-PCR, kit and cultivation to 182 clinical specimens, and the detection rates were 55.49％, 52.75％, 47.25％ and 39.01％ ,respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and nest-PCR were 89.6％ ,0.790, respectively;while those of the new method and cultivation were 83.5 ％ ,0.678, respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and the kit were 89.6％ ,0.792,respectively;and the correlation coefficient of these two methods was 0.923,P ＜ 0.001.Conclusion Compared with other methods, the new real-time PCR assay could be used to detect mycoplasma pneumoniae quickly and economically, with high sensitivity and specificity ,revealing great utility value on varied instrumentation platforms.

Objective To study the distribution of antinuclear antibodies ( ANAs) in a healthy population and the significance of using ANAs screening test in medical examination .Methods The ANAs were measured by indirect immunofluorescence assay ( IIF) .The Western blot assay was used to detect fif-teen specific antibodies against auto-antigens .Results 3519 out of all 25 110 subjects showed ANAs titers>1∶100 , and among them male and female subjects were respectively accounted for 1143 and 2376 .1489 out of all subjects had ANAs titers >1∶320 , and among them male and female subjects were respectively accounted for 406 and 1083 .The positive rates of ANAs at different titers showed significant differences be -tween male and female subjects .Among subjects with ANAs titers >1∶320 , the number of male subjects showed a steady increase with the age , while the percentage of female subjects reached to two peaks during the periods of puberty and menopause .The fifteen specific antibodies were detected in 659 out of 1489 sub-jects with ANAs titers>1∶320 and anti-Ro-52 (14.2%) accounted for the majority , followed by anti-M2 (12.7%) and anti-SSA (9.6%).Conclusion ANAs can be detected among healthy population of all ages, but their distribution varied with gender and age .ANAs screening test is necessary for medical exami-nation of healthy population , especially for female during period of puberty or menopause .The population with positive ANAs should be followed-up closely and educated for the prevention of autoimmune diseases .

Objective:To observe the clinical effect of deep insertion at Tianshu(ST 25)for colonic slow transit constipation(STC).Methods:120 cases of STC patients were randomly divided,60 cases in a deep insertion group,30 cases in an electroacupuncture group and 30 cases in a medication group by 2:1:1 ratio.The deep insertion group was treated with deep insertion at Tianshu(ST 25).The electroacupuncture group was treated with routine insertion at Tianshu(ST 25).The medication group was treated with oral administration of Lactulose oral liquid.The first voluntary defecation time,and constipation scores before the treatment,four weeks after the treatment and relevant scores of clinical symptoms were assessed in the three groups of the patients.Results:The scores of the clinical symptoms in improvement of constipation were better in the deep insertion group than in the electroacupuncture group and medication group,with differences in statistical significance(P<0.01).The unsuccessful numbers in the improvement of defecation and abdominal pain were also better in the deep insertion group than in the other two groups,and better in instant effect in the deep insertion.Conclusion:The improvement of STC clinical symptoms was better by deep insertion at Tianshu(ST 25)than by medication and routine acupuncture method at Tianshu(ST 25).