Objective:To analyze the mutation of T-cell and B-cell epitopes derived from HBV PreS-S protein in occult hepatitis B virus (OHBV) and investigate the biological mechanisms of occult hepatitis B virus infection (OBI) and HBsAb positive OBI.Methods:The PreS-S region of OBI samples were amplified by nested PCR, the products were sequenced and HBV genotypes were determined. The mutations of T-cell and B-cell epitopes derived from HBV PreS-S protein were analyzed and compared among groups of HBV genotypes and the presence of HBsAb. The affinity of the high frequency of T-cell epitope substitutions were analyzed by SYF PEITHI, the changes of antigenic characteristics of high frequency of B-cell epitope substitutions were analyzed by Ab Designer, Expasy ProtParam tool, Epitope Prediction and Analysis Tools.Results:The PreS-S region of HBV was amplified in 21 samples, including 4 HBsAb+ OBI B, 6 HBsAb-OBI B, 6 HBsAb+ OBI C, 5 HBsAb-OBI C. The mutation rates in PreS-S region of OBI were significantly higher than wild type HBV strains(OBI Bvs. WT B: 2.64%: 0.66%, P<0.001; OBI Cvs. WT C: 3.67%: 1.19%, P<0.001). The mutation rates of the immunoreactive area were significantly higher than non-immunoreactive area in OBI (OBI B: 3.57%: 1.86%, P=0.005; OBI C: 4.78%: 2.65%, P<0.001). The mutation rates of the immunoreactive and non-immunoreactive area in OBI C were higher than OBI B, but there was no statistically significant difference (immunoreactive area: 4.78%: 3.57%, P=0.107; non-immunoreactive area: 2.65%: 1.86%, P=0.142). The mutation rates of T-cell and B-cell epitopes of HBsAb-OBI were higher than HBsAb+ OBI, although there was no significant difference (HBsAb-OBI Bvs. HBsAb+ OBI B: 4.17∶3.01, P=0.303; HBsAb-OBI Cvs. HBsAb+ OBI C: 5.65∶4.26, P=0.207). The affinity analysis of 4 high frequency T-cell epitope substitutions, including T47A/K, S174N, L175S, V177A, showed that the changes of affinity of most mutation sites were not obvious; the antigenicity analysis of 3 high frequency B-cell epitope substitutions, including G73S, K122R, I126M/T, did not show noticeable changes and the hydrophilicity, surface accessibility of some mutation sites were even better than wild strain. Conclusions:The mutation rates in PreS-S region of OBI were significantly higher than wild type HBV strains. The mutation rates of the immunoreactive area were higher than non-immunoreactive area in OBI. The variant activity of OBI C was higher than OBI B. The mutations of OBI might occur randomly and were not selected by antibody pressure. Single epitope and multi-epitopes combinational mutations might be a reason for OBI.

【Objective】 To analyze the composition of urinary calculi in Longnan, Gansu province, and the relationship between the composition and clinical characteristics of patients, so as to provide reference for the prevention and treatment of this disease. 【Methods】 The composition of 500 cases of urinary calculi hospitalized in our department during Apr. 2021 and Feb.2023 were analyzed using the infrared spectrum stone composition analyzer. The clinical characteristics of the patients were evaluated and analyzed. 【Results】 The male-to-female ration of patients was 2.70∶1. Most patients aged 21 to 60 years (437, 87.4%). Most cases were ureteral calculi (72.8%), followed by renal calculi (22.2%), and bladder calculi (5.0%). There were 166 cases of calcium oxalate calculi, 293 cases of calcium oxalate + carbonated apatite calculi, 24 cases of calcium oxalate + carbonated apatite + hydroxyl apatite calculi, 4 cases of calcium oxalate + calcium bicarbonate calculi, 7 cases of urate calculi, 6 cases of carbonated apatite + struvite calculi. Oxalate calculi were the most common in all age groups, and urate calculi were the most common in the 21 to 40 age group. Calcium oxalate calculi were most common in the ureter (127, 76.5%), significantly higher than in other sites (χ²=3.222, P=0.020). Calcium oxalate + calcium hydrogen phosphate calculi was the least common in the bladder, significantly different from the other parts (χ²=2.092, P=0.037). Magnesium ammonium phosphate hexahydrate and/or calcium carbonate or calcium oxalate calculi were the most common in the kidney (50.0%), significantly different from the other parts (χ²=9.448, P=0.007). 【Conclusion】 In Longnan area, the incidence of urinary calculi is significantly higher in male than in female. Ureteral calculi are mainly composed of calcium oxalate + carbonated apatite and calcium oxalate. According to different risk factors, individual prevention programs should be developed.

【Objective】 To investigate the distribution of plasma donors with high titer neutralizing antibodies against human cytomegalovirus (HCMV) in the general plasma donor population. 【Methods】 920 plasma samples of Taibang were tested in April 2014 to investigate the distribution of anti-HCMV neutralizing antibodies. After further testing of mixed plasma, the threshold for screening plasma was determined. From October 2019 to May 2020, neutralizing anti-HCMV in 40 078 plasma samples from 11 plasma stations in Shandong province were screened by the microcytopathic method (modified high-flux neutralization test method). The proportion of neutralizing anti-HCMV enriched in high titer and the distribution in the donor population were analyzed by SPSS 26 and Minitab19 analysis software. 【Results】 Among 920 samples, 73.26%, 0.43%, and 8.69% of them had neutralization titer<1∶15, ≥1∶60 and ≥1∶30, respectively. The neutralization titer of mixed plasma was detected, and 1∶30 was determined as the high titer. The yielding rate of high titer neutralizing anti-HCMV in Shandong was 9.06% (3 633/40 078). The proportion of plasma donors with high-titer neutralizing anti-HCMV in the donation population from plasma stations was 4.95%~13.03% (9.06±2.07) %. The proportion of plasma donors with high-titer neutralizing anti-HCMV by gender was 15.67% (2 185/13 951) in women and 5.54% (1 448/26 127) in men(P<0.05). 【Conclusion】 There was a certain proportion of plasma donors wiht high titer neutralizing anti-HCMV in the population of plasma donors in Shandong, and they can constantly serve neutralizing anti-HCMV to ensure the production of anti-HCMV immunoglobulin preparations.

Objective To analyze the current status of exercise and exercise self-efficacy (ESE) and its influencing factors in middle-aged and elderly patients with high blood pressure, and to provide a basis for the intervention of patients with hypertension in the community. Methods The stratified random sampling method was used to select 546 patients with hypertension who had received hypertension management for one year in the community. The general condition, exercise condition, and ESE scores were collected using survey questionnaires. Chi-square test was applied to analyze the difference of exercise efficacy score among different characteristic objects, and analysis of regression method was applied to analyze the influencing factors of exercise self-efficacy. Results A total of 546 people were surveyed in this study, of which 67.03% of hypertension patients participated in exercise. The main exercise program was walking healthily (87.43%). A total of 262 people (47.99%) had strong performance on ESE score, while 284 people (52.01%) had weak self-efficacy. The univariate unconditional logistic regression analysis found that there were significant differences in the distribution of educational level, family monthly income, years of hypertension, whether they received the guidance from family doctors, whether they exercised and whether they warmed up before exercise (P<0.05). The multivariate unconditional logistic regression analysis showed that educational level of junior high school and low average monthly household income were the risk factors affecting exercise efficacy, and the guidance of family doctors, exercise, and hypertension less than 5 years were the protective factors affecting exercise efficacy. Conclusion The exercise self-efficacy of middle-aged and elderly patients with hypertension in this community is at a medium level, and family doctors should carry out targeted interventions based on influencing factors to improve exercise self-management ability.

Objective:To analyze the etiological characteristics and molecular epidemiological correlation of five cases of typhoid fever during the same period in yantai city.Methods:Six S. Typhis strains were isolated from 5 typhoid patients and epidemiological samples in Yantai city in 2018. The onset time of the cases were from May 26, 2018 to July 24, 2018, distributed in Shuidao Town of Muping District, Dengzhou Street of Penglai District, Donglai Street of Longkou District, Wenhua Street of Muping District and Fulaishan Street of zhifu District. S. Typhis strains were analyzed by conventional bacterial isolation method and XbaⅠ/BlnⅠ double-enzyme digestion pulse-field gel electrophoresis (PFGE). Meanwhile, ViaB virulence gene detection and 27 common antibiotics sensitivity tests were conducted to study the etiology of S. Typhis.Results:Six strains of S. Typhi were isolated from 5 patients and the domestic egg of one patient, which were divided into 4 PFGE patterns by PFGE-XbaⅠ and PFGE-BlnⅠ and among which 3 strains had the same PFGE patterns.One multi-drug resistant strain (foreign patient), one single-drug resistant strain (patient with a history of provincial retention), and one completely sensitive strain were detected. The three strains of the same PFGE pattern exhibit the same drug-sensitive phenotype which were intermediate against aminoglycosides and quinolones and susceptibility against the other antibiotics.All of the strains carried the ViaB virulence factor except the strain from the foreign patient.Conclusion:Local S. Typhi is susceptibility or intermediate against antibiotics commonly used in clinic.Sporadic cases of typhoid fever and typhoid imported infections still need attention.

Objective:To analyze the etiological characteristics and molecular epidemiological correlation of five cases of typhoid fever during the same period in yantai city.Methods:Six S. Typhis strains were isolated from 5 typhoid patients and epidemiological samples in Yantai city in 2018. The onset time of the cases were from May 26, 2018 to July 24, 2018, distributed in Shuidao Town of Muping District, Dengzhou Street of Penglai District, Donglai Street of Longkou District, Wenhua Street of Muping District and Fulaishan Street of zhifu District. S. Typhis strains were analyzed by conventional bacterial isolation method and XbaⅠ/BlnⅠ double-enzyme digestion pulse-field gel electrophoresis (PFGE). Meanwhile, ViaB virulence gene detection and 27 common antibiotics sensitivity tests were conducted to study the etiology of S. Typhis.Results:Six strains of S. Typhi were isolated from 5 patients and the domestic egg of one patient, which were divided into 4 PFGE patterns by PFGE-XbaⅠ and PFGE-BlnⅠ and among which 3 strains had the same PFGE patterns.One multi-drug resistant strain (foreign patient), one single-drug resistant strain (patient with a history of provincial retention), and one completely sensitive strain were detected. The three strains of the same PFGE pattern exhibit the same drug-sensitive phenotype which were intermediate against aminoglycosides and quinolones and susceptibility against the other antibiotics.All of the strains carried the ViaB virulence factor except the strain from the foreign patient.Conclusion:Local S. Typhi is susceptibility or intermediate against antibiotics commonly used in clinic.Sporadic cases of typhoid fever and typhoid imported infections still need attention.

Objective:To explore the clinical characteristics, genotypes and long-term outcomes of individuals with 3-methylglutaconic aciduria.Methods:The clinical features, biochemical data, genetic test results and treatment outcomes of six children with 3-methylglutaconic aciduria admitted to the Department of Endocrinology, Genetics and Metabolism, Xinhua Hospital from February 2017 to February 2019 were retrospectively analyzed and the Gesell developmental diagnosis schedule was performed to evaluate the development of four patients.Results:Among 6 children with 3-methylglutaconic aciduria 2 were males and 4 were females.Four cases had 3-methylglutaconic aciduria type Ⅰ and 2 cases had 3-methylglutaconic aciduria with deafness,encephalopathy, and Leigh-like syndrome. Five of 6 patients were detected by newborn screening among whom 4 remained asymptomatic, and only one had a postmortem diagnosis. Among them, 4 patients remained asymptomatic, while two presented with clinical symptoms such as jaundice and dyspnea and the age of disease onset was 1 and 2 days respectively. The concentration of 3-methylglutaconic acid in urine of all affected individuals was between 22.38 and 77.09 mmol/molCr, which was above the normal value. Genetic tests were performed for all patients. Eleven variants were identified in 2 genes, of which 10 variants were novel and only c.442C>T p.(R148X) has been previously reported; Seven variants (c.656-2delA, EX5-EX6 Del, c.942+3A>G, c.373C>T p.(R125W), c.895-3C>G, c.667C>T p.(R223X) and c.894+5G>A) were in AUH gene. The others (c.548G>A p.(R138Q), c.442C>T p.(R148X), c.1339C>T p.(R447X) and c.973dupA p.(M325Nfs*5) were in SERAC1 gene. After being treated with leucine diet restriction and L-carnitine, 4 patients with AUH gene variation who were from asymptomatic phase developed normally, whereas those 2 patients with SERAC1 gene variation had a poor prognosis. During the follow-up, 2 patients exhibited varying degrees of psychomotor retardation, the rest had normal course of development.Conclusions:There are significant clinical heterogeneities among individuals with 3-methylglutaconic aciduria. The most common pathogenic variants are splicing variations, followed by nonsense, missense and frameshift mutations. Leucine-free diet and oral L-carnitine therapy are effective for some patients. Newborn screening is essential for early diagnosis and improvement of prognosis.

Objective To analyze the visual acuity and refractive status of kindergarten children and primary and middle school students in Xinhong Community of Shanghai from 2015 to 2018, and to provide basis and reference for the prevention and control of students^' myopia. Methods The data of 3 904 person-times of complete visual acuity and refraction records from 2015 to 2018 was derived from the ^"Shanghai Residents^' Eye Health Information Service System^". Results From 2015-2018, the total detection rate of poor vision of kindergarten children and primary and middle school students was 60.86%, and the detection rate of poor vision in kindergarten, primary school and middle school was 29.90%，62.63% and 87.26% respectively，showing an increasing trend with the education stage (χ²=727.206，P<0. 05). The detection rate of poor vision in boys was 57.31%, and that of girls was 64.29% (χ²=19.949, P<0.05). According to the classification of the degree of visual impairment, the peak stage of mild poor vision was in grade one to three of primary school, the peak stage of moderate poor vision was in grade four and five of primary school, the peak stage of severe poor vision was in middle school, and the highest peak of poor vision in second grade of middle school, accounting for about 70.85%. The total detection rate of poor vision with glasses reached 18.21%. The total detection rate of myopia in refractive state was 54.79%, and the detection rate of myopia in kindergarten, primary school and middle school were 16.27%,56.61% and 88.28% respectively, showing an increasing trend with the education stage (χ²=1099.978, P<0.05). In terms of growth rate, the increase in myopia in the first grade of primary school was the largest. Conclusion The myopia rate of primary and secondary school students was higher than that of kindergarten children, and showed an increasing trend with the education stage. The critical period of myopia prevention and control should be advanced to the kindergarten stage.

Objective To compare the value of presepsin,procalcitonin (PCT) and C-reactive protein (CRP) in differentiating different types of pathogenic bacteria in septic patients.Methods A prospective study was conducted to collect 322 septic patients who met the diagnostic criteria of "sepsis 3.0"in the Emergency ICU of the First Affiliated Hospital of Dalian Medical University from July 2016 to January 2018.According to the results of blood culture,patients were divided into the positive blood culture group (n=114) and negative blood culture group (n=208).Patients in the positive blood culture group were further divided into four subgroups:Gram-positive coccus (G+),Gram-negative bacilli (G-),mixed bacteria,and fungi groups.Healthy volunteers were selected as the control group (n=45).The differences in presepsin,PCT and CRP levels were compared among the groups,and the curves of the subjects' working characteristic curve (ROC) were drawn.Results Presepsin,PCT,and CRP were significantly increased in the positive blood culture and negative blood culture groups compared with the control group (all P＜0.05);Presepsin and PCT were significantly higher in the positive blood culture group than those in the negative blood culture group (both P＜0.05).There was no significant difference in Presepsin among the four subgroups in the positive blood culture group (all P＞0.05),but PCT was significantly higher in the G-and mixed bacteria groups than that in the G+ and fungi groups (all P＜0.05).Presepsin predicted a positive blood culture with area under ROC curve of 0.680,which was higher than PCT (AUC=0.599).Conclusions Presepsin is more valuable than PCT in early predicting positive blood culture in septic patients,but only PCT has an ability to differentiate pathogenic bacteria in septic patients with positive blood culture.It suggested that a combination of Presepsin and PCT should be more meaningful in clinical practice.

Objective: To analyze the pathological features and their influence on the clinical outcome of non-nasopharyngeal EBV-associated carcinomas. Methods: One hundred and twenty cases of non-nasopharyngeal EBV-associated carcinoma confirmed by in situ hybridization were identified at Zhejiang Cancer Hospital from January 1, 2006 to May 1, 2018, and the clinicopathological data were collected and analyzed using Kaplan-Meier survival analysis, Cox univariate and multivariate analysis. Results: One hundred and twenty cases were involved in the study; the male to female ratio was 1∶1; patients′ age range was 24 to 89 years (median 50 years). The primary sites were large parotid glands (62 cases), lung(26 cases), stomach(15 cases), and others (oral, oropharynx, larynx, cervix, liver; totally 17cases). Non-nasopharyngeal EBV-associated cancer could be divided into two histological types according to the amount of interstitial lymphocytes: type Ⅰ was "lymphoepithelial-like carcinoma" and rich in stromal lymphocytes; type Ⅱ lacked lymphocytic infiltration. Ninety-eight primary tumor samples could be classified morphologically: 43 cases were as type Ⅰ and 55 cases as typeⅡ; the distribution of type Ⅰ was 57.4% (27/47) in large parotid glands, 20.8% (5/24) in lung, 4/13 in stomach, and 7/14 in other sites. Complete treatment and survival data were obtained for 114 patients. According to the TNM staging criteria of WHO, 52 patients were at early stages (Ⅰ-Ⅱ) and 62 were at advanced stages (Ⅲ-Ⅳ); 102 patients underwent surgery. Seventy-four patients received adjuvant chemotherapy before or after surgery, and 52 patients received local radiotherapy. Kaplan-Meier survival analysis showed that patients with type Ⅱ EBV-associated carcinoma had a worse prognosis than patients with type Ⅰtumors (P=0.010 2). In addition, vascular invasion(P=0.021 8),neural recidivism(P=0.000 1),advanced stage(P=0.017 1),lymph node metastasis (P=0.005 0) and chemotherapy (P=0.013 2) were poor prognostic factors; female patients had better survival than male (P=0.028 4). Cox multivariate regression analysis found that lymph node metastasis (95%CI: 1.489-13.830, P=0.007 6) and neural recidivism (95%CI: 1.228-6.544, P=0.014 7) were independent adverse prognostic factors. Cox multivariate regression analysis after stratification by site revealed that radiotherapy was a preferable prognostic factor for EBV-associated carcinoma of the large salivary glands (95%CI: 0.003-0.569, P=0.016 8). Conclusion EBV associated carcinoma can be divided into two types, for which type Ⅰ was with abundant interstitial lymphocytes and type Ⅱ was lack of interstitial lymphocytes. TypeⅡ EBV-associated carcinoma has a worse prognosis than type Ⅰ. Radiation therapy can prolong the survival time of patients with primary EBV-associated carcinoma of large salivary glands.