Objective To investigate the effect of age on the incidence of cirrhosis and liver cancer in patients with chronic hepatitis B.Methods 279 patients with chronic hepatitis B were divided into the senior group and the younger group according to the age of the patients.The cumulative incidence of cirrhosis and liver cancer during 25 years of follow-up was calculated by using SPSS and R language through the long-term follow-up of HIS system,and the risk factors were analyzed by multivariate logistic regression.Results During follow-up,24 cases developed cirrhosis and 12 cases developed liver cancer.The cumulative incidence of liver cirrhosis was 1.5%,2.1%,5.4%,11.6%and 15.5%in the 5-year,10-year,15-year,20-year and 25-year group,and 5.5%,9.8%,22.9%,29.0%and 52.1%in the elderly,respectively.The difference between the younger age group and senior age group was statistically significant(P<0.001).A total of 2 risk factors(age and follow-up time)were included in the regression model.Two cases in the younger group developed into liver cancer after 17 and 21 years of follow-up,respectively.The cumulative incidence rates at 5,10,15,20 and 25 years were 1.8%,3.8%,18.5%,21.8%and 26.7%.A total of five factors(initial age,HBV-DNA load,HBV-DNA turned negative before the end-point,follow-up time,and sex)were included in the regression model.Conclusions The incidence of cirrhosis and liver cancer in CHB patients aged≥40 years,especially in male patients,is significantly higher than younger CHB patients.Timely initiation of antiviral therapy can delay disease progression and reduce the incidence of termi-nal liver disease.Whether antiviral therapy should be initiated for people aged 30 to 40 years remains to be studied.

Objective To establish and optimize a method for simultaneous determination for the contents of sodium,potassium,calci-um and magnesium ions in serum using microwave digestion combined with ion chromatography,and evaluate its performance.Methods Serum samples were pretreated by microwave digestion,and four cations were quantitatively determined using ion chromatography.The linearity,precision,recovery and accuracy of the established method were verified with reference to the EP6-A and EP15-A3 docu-ments issued by the Clinical and Laboratory Standards Institute(CLSI).Results The imprecisions of determinations for sodium,po-tassium,calcium and magnesium ions expressed as coefficient of variation(CV)were 0.10％to 0.36％,0.15％to 0.48％,0.22％to 0.87％and 0.21％to 0.73％,respectively.The recoveries with adding standard of the four cations were between 97.3％and 103.0％.Referred to RELA2020 and RELA2021,the deviations of the analysis results were-0.57％to-0.14％,-0.84％to-0.16％,-1.82％to-0.37％,and-0.60％to 0.34％for sodium,potassium,calcium,and magnesium,respectively,and all the comparisons were pas-sed.The limits of detection/quantification were 0.017 5/0.058 3 mmol/L,0.000 5/0.001 7 mmol/L,0.009 9/0.032 9 mmol/L and 0.001 1/0.003 5 mmol/L for the four cations,indicating that after pretreatment,no significantly interfere from other impurities for the results of determination was found.Conclusion A method for simultaneous determination of sodium,potassium,calcium and magnesi-um ions in serum was successfully established and optimized using microwave digestion combined with ion chromatography.The method showed good precision and accuracy,simple operation,short time consumption and low cost,thus it could be used for quality improve-ment of clinical electrolyte testing in serum.

Objective:To develop a candidate reference method for cortisol in human serum by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS).Methods:An isotope-labeled internal standard was added to samples, followed by alkalizing with sodium carbonate solution and liquid-liquid extraction with n-hexane/ethyl acetate. Isoelution with a methanol aqueous solution was employed for liquid chromatographic separation, while ESI in the positive ion and multiple reaction monitoring mode were used for mass spectrometry. The volume of sample, standard, and internal standard solutions were all controlled by weighing, and the results were calculated by bracketing method. The accuracy, precision, specificity, linearity, LOD and LOQ of this method were evaluated referring to the CLSI C62-A and C50-A guidelines and the domestic expert consensus documents.Results:The method demonstrated excellent accuracy and precision with the bias of ERM-DA192, ERM-DA193 and RELA 2021-2023 all being less than 2%. The recovery of added cortisol ranged from 98.2% to 101.1%. Both intra- and inter-assay imprecisions was <2%. The method was free from interference by structural analogue and showed a good linearity in the range of 25-1 600 nmol/L. The LOD and LOQ were 0.5 nmol/L and 1.0 nmol/L, respectively. A cortisol assay kit (chemiluminescence immunoassay) traced to this candidate reference method was used to determine 46 clinical serum samples concurrently, and the two methods exhibited good correlation.Conclusions:A candidate reference method for the determination of cortisol in serum was established, demonstrating high sensitivity, good repeatability and accuracy. This method can serve as a reference for the measurement traceability and accuracy evaluation of routine methods.

Objective:To explore the effect of oral nutritional supplements (ONS) during labor on delivery outcomes in low-risk pregnant women and the risk factors of cesarean section.Methods:A retrospective analysis was conducted in a total of 206 full-term pregnant women with singleton and cephalic presentation at the delivery center in our hospital from March 15th to May 15th, 2022. Standard diet education was given to all those women by midwives. Pregnant women who received the enteral nutrient solution prepared by the Department of Clinical Nutrition during labor were in the ONS group ( n = 110), while those who did not were in the control group ( n = 96). The baseline characteristics and delivery outcomes were compared between the two groups, and the risk factors of cesarean section were also analyzed. Results:There were no significant differences in terms of maternal age, height, baseline weight, baseline body mass index (BMI), weight gain during pregnancy, prenatal BMI, gestational week, intraspinal labor analgesia, oxytocin, gastrointestinal intolerance, neonatal length, and weight between the two groups ( P > 0.05). However, the total oral energy intake during labor in the ONS group was significantly higher than that in the control group ([1 349.99± 569.51] kJ vs [249.59 ± 455.19] kJ, P < 0.01). The rate of vaginal delivery in the ONS group was significantly higher than that in the control group (93.6% vs 81.3%, P = 0.01), and the duration of the first stage of labor ([487.06 ± 232.94] min vs [416.17 ± 191.13] min, P = 0.03) was also significantly longer in the ONS group. There were no significant differences between the two groups in terms of the duration of the second and third stages of labor, perineal laceration, cervical laceration, vaginal laceration, amount of bleeding, hospital stay, and Apgar score after birth ( P > 0.05). Multivariate logistic regression analysis showed that maternal age was a risk factor for changing to cesarean section in women with low-risk pregnancies ( OR 1.20, 95% CI 1.03 to 1.40, P = 0.02), while ONS during labor ( OR 0.31, 95% CI 0.11 to 0.85, P = 0.02) and intraspinal labor analgesia ( OR 0.10, 95% CI 0.04 to 0.26, P < 0.01) were protective factors. Conclusions:ONS during labor can significantly increase the vaginal delivery rate, without increasing the incidence of gastrointestinal intolerance in low-risk pregnant women. Maternal age is a risk factor for cesarean section while ONS during labor and intraspinal labor analgesia are protective factors in women with low-risk pregnancies.

Objective:To investigate the effect of astragaloside Ⅳ on the apoptosis of thyroid cells in Hashimoto's thyroiditis(HT)rats and Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing kinase 2(ROCK2)pathway.Methods:The HT rat model was induced by subcutaneous injection of thyroglobulin combined with high iodine drinking water and randomly divided into model group,astragaloside(80 mg/kg)group,Rhosin(RhoA inhibitor,40 mg/kg)group,astragaloside Ⅳ(80 mg/kg)+ Rhosin(40 mg/kg)group(12 rats in each group),another 12 SD rats were selected and drank water normally and injected the same dose of saline subcutaneously as control group.After the drugs were grouped and processed,the serum anti-thyroglobulin antibody(TGAb),anti-thyroid peroxidase antibody(TPOAb)levels and the inflammatory factors IL-6,IL-17,IL-1β contents were measured by ELISA kits;the pathological changes of thyroid tissue in each group were detected by hematoxylin-eosin(HE)staining;the apopto-sis rate of rat thyroid cells in each group were detected by TUNEL staining;the expressions of RhoA/ROCK2 pathway proteins in thy-roid tissues of rats in each group were detected by Western blot.Results:Compared with the control group,the thyroid follicles in the model group had abnormal structure,some atrophy or disappearance,disordered arrangement,surrounding inflammatory cell infiltra-tion,and obvious pathological damage to the thyroid tissue,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,and thyroid tissue RhoA and ROCK2 protein expression levels were significantly increased(P<0.05);compared with model group,the pathological damage of the thyroid tissue of rats in the drug intervention group were reduced,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,and thyroid tissue RhoA and ROCK2 protein expression levels were decreased(P<0.05);compared with astragaloside Ⅳ group and the Rhosin group respectively,the pathological damage of the thyroid tissue of rats in the astragaloside Ⅳ+Rhosin group were further reduced,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,thyroid tissue RhoA and ROCK2 protein expression levels were decreased(P<0.05).Conclusion:Astragaloside Ⅳ may down-regulate the expression of RhoA/ROCK2 pathway to reduce the inflammatory injury of thyroid tissue,inhib-it thyroid cell apoptosis,and improve the symptoms of HT in rats.

[摘 要] 目的：检测食管鳞状细胞癌（ESCC）患者血清中高迁移率族蛋白B1（HMGB1）和吲哚胺-2，3-双加氧酶（IDO）的表达水平并探讨两者与临床病理特征及淋巴细胞亚群的相关性。方法：选取2021年3月至2022年8月在河北医科大学第四医院初次住院治疗的95例ESCC患者作为ESCC组，另选取40例健康体检人群作为对照组。ELISA法检测全部研究对象的血清HMGB1和IDO水平及不同组ESCC细胞培养上清中HMGB1、IDO和p65水平，流式细胞术检测全部研究对象外周血淋巴细胞亚群水平。WB法检测仅敲低HMGB1基因表达或敲低HMGB1后再加入NF-κB信号通路激活剂对ESCC细胞HMGB1、IDO和p65表达的影响。结果：ESCC组患者血清HMGB1和IDO水平明显高于对照组（均P<0.01）；血清HMGB1和IDO表达水平升高是ESCC临床进展的独立危险因素（均P<0.01），二者联合检测对ESCC临床进展预测价值更高（P<0.01）；血清HMGB1和IDO与ESCC患者的T分期、N分期和临床分期有明显关联（均P<0.05）； ESCC组患者血清HMGB1与外周血CD3+ T细胞、CD4+ T细胞、B细胞和NK细胞绝对计数值呈显著负相关，而与Treg细胞百分率呈显著正相关（均P<0.05），血清IDO与外周血CD3+ T细胞百分率和绝对计数值、CD4+ T细胞百分率和绝对计数值、CD8+ T细胞和B细胞绝对计数值呈显著负相关，而与Treg细胞百分率呈显著正相关（均P<0.05）；血清HMGB1和IDO表达水平呈显著正相关（P<0.01）。si-HMGB1组KYSE30和ECA109细胞及其培养上清液中IDO和p65表达水平明显低于si-NC组和si-HMGB1+PMA组（均P<0.05）。结论：血清HMGB1和IDO与ESCC临床进展和机体免疫功能密切相关，具有成为ESCC肿瘤标志物和免疫治疗新靶点的潜力。HMGB1可能通过NF-κB信号通路促进IDO表达，进行双靶点联合治疗可能会取得更好的疗效。

Objective:To explore the value of Neoseq in screening and diagnosis of neonatal fatty acid oxidation disorders (FAOD).Methods:A retrospective case-control study was conducted on 163 500 live births in Changzhou city from April 2015 to April 2021. The following two models were adopted for FAOD screening and diagnosis. (1) Traditional mode: Heel blood samples were obtained from all subjects for initial screening using tandem mass spectrum (TMS), followed by next-generation sequencing (NGS) and other differential diagnostic testings for those with positive results. (2) Neoseq: Neoseq was performed on the true positive, negative and false positive cases according to the traditional mode screening results. The detection rate, additional discovery, reporting period, and other parameters of the two models for FAOD were described and compared.Results:(1) Detection and diagnosis of FAOD: A total of 18 confirmed cases of FAOD were detected through the traditional model, with an incidence of 1/9 083 in Changzhou city. The positive rate was 0.55% (907/163 500) for initial TMS and 0.04% (73/163 500) for the second. The positive predictive value was 2.0%(18/907), with a false positive rate of 98%(889/907) in the initial screening. (2) The results of Neoseq: ①Pathogenic mutations were detected in 16 of the 18 confirmed cases, and the coincidence rate of mutation sites between the two methods was 16/18. The other two confirmed cases were missed diagnosed by Neoseq, including one β-ketothiolase deficiency with only one detected pathogenic mutation and one medium-chain acyl-CoA dehydrogenase deficiency without any detected pathogenic mutation. ②No pathogenic mutations were detected in the 57 false-positive cases by Neoseq. ③Among the 100 negative cases in initial screening, DUOX2 heterozygous mutation, and MTTL1 hemizygous mutation were detected in one case each. ④The median period of results reporting was 43.5 d (28-104 d) for the traditional mode and 12 d (10-15 d) for the Neoseq mode. Conclusions:Neoseq has a high detection rate for FAOD. Combined with TMS screening, Neoseq reduces the false-positive rate of biochemical screening, rapidly identifies genetic causes by shortening the results waiting time and covers diseases that couldn't be detected by traditional biochemical methods.

Objective:To assess the differences of the 2016 and 2009 edtion guidelines on diastolic dysfunction in sepsis patients.Methods:A single-center, prospective study was conducted. The relevant information of sepsis patients in Intensive Care Unit (ICU) were analyzed from October 2016 to January 2019. Patients’ transthoracic echocardiography at the first 24 h and 3rd day of their admission and left ventricular diastolic dysfunction were stratified according to the 2009 and 2016 edition guidelines. Patients’ characteristics, arterial blood gas analysis, and blood biochemical indexes were recorded at the first 24 h of the ICU admission. Additionally, the following information were retrieved during ICU stay: site of infection, frequency of adrenaline and dobutamine, maximal dose of norepinephrine, use of hydrocortisone, invasive mechanical ventilation and renal replacement therapy. The rank-sum test of two independent samples was used to compare the differences in the diagnosis of left ventricular diastolic dysfunction.Results:A total of 196 patients with sepsis or septic shock were screened, and 86 patients were excluded. Finally, clinical data of 110 patients were included in the analysis.The median time of the first ultrasound examination in ICU was 17 h. Among the patients with different diastolic function severity in baseline data analysis, only age was significantly different. According to the 2016 edition guidelines, 43 (39%) of 110 patients had diastolic dysfunction and another 30 (27%) had indeterminate diastolic dysfunction within 24 h of ICU admission. According to the 2009 edition guidelines, 40 (36%) patients had diastolic dysfunction and 58 (53%) patients had indeterminate diastolic dysfunction. The diagnosis of left ventricular diastolic dysfunction of different grades was significantly different between the 2016 and 2009 edition guidelines ( Z=4.92, P<0.01). According to the 2016 edition guidelines at the 3rd day of ICU admission, 52 (47%) patients were diagnosed with diastolic dysfunction and 18 (18%) were diagnosed with indeterminate diastolic dysfunction; According to the 2009 edition guidelines, 50 (46%) of these patients were diagnosed with diastolic dysfunction and 45 (41%) had indeterminate diastolic function. Similarly, there was a large difference in diagnosis ( Z=4.60, P<0.01). Subgroup analysis of patients with normal systolic function (ejection fraction > 50%) showed that the diagnosis of left ventricular diastolic dysfunction were significantly different at the first 24 h and the 3rd day of ICU admission ( Z=4.34, P<0.01 and Z=5.71, P<0.01). Conclusions:The 2016 edition guidelines identify a significantly higher incidence of dysfunction in patients with severe sepsis and septic shock compared to the 2009 edition guidelines. Although the 2016 edition guidelines seem to be an improvement, issues remain with the application of guidelines using traditional measures of diastolic dysfunction in this cohort.

Objective To investigate the prognostic impact of left ventricular diastolic dysfunction in septic shock patients in intensive care unit (ICU). Methods A single-center, prospective tudy was conducted. The relevant information of septic shock patients who underwent an echocardiographic assessment during the initial management were analyzed in ICU of Harrison International Peace Hospital Affiliated to Hebei Medical University from June 2016 to October 2017. Hemodynamics were evaluated using transthoracic echocardiography within the first 24 h of shock, and LV diastolic dysfunction is present if the available parameters meet the cutoff values. Patients' characteristics, arterial blood gas analysis, and blood biochemical indexes were recorded in the first 24h at ICU admission. Additionally, the following information were retrieved during ICU stay: site of infection, frequency of adrenaline and dobutamine, maximal dose of norepinephrine, use of hydrocortisone, invasive mechanical ventilation and renal replacement therapy. A multivariate analysis was performed to determine independent risk factors associated with septic shock mortality, and the receiver operating characteristic curve (ROC) was drawn to evaluate the predictive value of all risk factors on the outcome of patients. Results Among the 138 patients with septic shock hospitalized in the ICU during the study period, 102 patients were enrolled [76 men (75％); aged 64 ± 12 years] and 92 of them (90％) were mechanically ventilated. Thirty patients died in ICU with a mortality rate of 29％. LV diastolic dysfunction was observed in 32 patients (31％). No significant inter-group difference in infection characteristics was found. The death group had significantly higher APACHE Ⅱ score, SOFA score, lactate and creatinine levels than the survival group. Similarly, the death group received a greater dose of vasopressors and required more frequent administration of epinephrine than the survivors. While the dead group exhibited slightly yet significantly larger RV cavity, which was reflected by a significant greater RVEDA/LVEDA ratio (0.77 ± 0.43 vs 0.59 ± 0.17) with no association with paradoxical septal motion. Mean value of lateral e′ maximal velocity was significantly lower in the death group compared with the survival group(10.50 ± 3.80)cm/s vs (12.9 ± 4.6)cm/s. The proportion of LV diastolic dysfunction tended to be higher in the death group than in the survival group (14/30 vs 18/72). In the multivariate analysis, the maximal dose of norepinephrine (OR=1.69, 95％CI:1.05-2.79), the SOFA score(OR=2. 24,95％CI:1.32-3.35) ,e′ lateral maximal velocity (OR=2.23, 95 ％ CI:1.01-5.57), lactate(OR=2.36, 95％CI:1.25-5.43) and LV diastolic dysfunction(OR=1.32,95％CI:1.11-3.15) were significantly associated with septic shock mortality. The area under ROC curve of lateral e′ maximal velocity for predicting death of septic shock patients was 0.63 (95％CI: 0.54-0.78) and the cut-off value was 8.9 cm/s, with a sensitivity and specificity of 76％ and 54％, respectively. Conclusions The present study suggests that LV diastolic function might be associated with ICU mortality in patients with septic shock. A multicenter prospective study assessing a large cohort of patients using serial echocardiographic examinations remains to confirm the prognostic value of LV diastolic dysfunction in septic shock patients.

Objective To examine the effect of adolescent intermittent ethanol exposure (AIE) on ethanol-induced conditioned place preference (CPP) and anxiety-like behavior in adolescent mice. Methods In experiment 1, adolescent male C57BL/6 mice at 4 weeks of age were randomly divided into AIE group and NS group (n=10 for each group). The binge drinking model was established by AIE (3 g/kg, 25%). The alcohol reward was evaluated using the ethanol-induced CPP paradigm (2 g/kg, 20%). In experiment 2, the anxiety-like behavior of adolescent male C57BL/6 mice were assessed using the elevated plus maze (EPM) test, and the animals were then allocated into high-anxiety mouse (HAM) and low-anxiety mouse (LAM) groups based on the percentage of open arm time (OT%). HAM and LAM were randomly divided into AIE group and NS group (n=8～10 for each group) with random number method, respectively. Then, anxiety-like behavior in four groups was measured again using the EPM test. Results In experiment 1. Ethanol preference (116.1± 12.9)s vs. (70.8±14.8)s, P=0.035) was significantly higher in AIE group relative to NS group. However, In experiment 2. The alteration in anxiety-like behaviors was not significant in either HAM-AIE or LAM-AIE groups (all P＞0.05). Conclusions AIE reinforces ethanol-induced CPP but does not affect the anxiety-like behavior in adolescent mice, suggesting that AIE may not play a role in anxiety-like behavior.