Table of Content

30 July 2023, Volume 38 Issue 7

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Expert consensus on the clinical application of AFP，AFP-L3% and DCP using GALAD and GALAD-like models in hepatocellular carcinoma

Clinical Laboratory Society of Chinese Association for Rehabilitation Medicine , Molecular Diagnostics Society of Shanghai Medical Association , Tumor Immunology Branch of Shanghai Society for Immunology , Yueyang Hospital of Integrated Traditional Chinese and Western Medicine of Shanghai University of Traditional Chinese Medicine , Shanghai Center for Clinical Laboratory, Clinical Laboratory Society of Chinese Association of Integrative Medicine , Clinical Laboratory Society of Shanghai Anticancer Association , Tumor Markers Society of Shanghai Anticancer Association

2023, 38(7):  607-623.  DOI: 10.3969/j.issn.1673-8640.2023.07.001

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Primary hepatocellular carcinoma （HCC） is a prevalent world-wide malignancy. Half of the newly developed HCC occurs in China. Optimizing the strategies for high-risk surveillance and early diagnosis is pivotal for improving 5-year survival. Constructing the scientific non-invasive determination technologies feasible for medical and healthcare institutions is a key route for elevating the efficacies in HCC diagnosis and follow-up. Based on the international and Chinese guidelines，expert consensus statements，literatures and evidence-based clinical practice experiences，this consensus published by the Clinical Laboratory Society of Chinese Association for Rehabilitation Medicine puts forward the clinical implications，application subjects，determination techniques and result interpretations for GALAD and GALAD-like models based on alpha-fetoprotein （AFP），alpha-fetoprotein heterogeneity-L3%（AFP-L3%），and des-gamma-carboxy prothrombin（DCP）. The compile of this consensus statement aims to address and push the reasonable model application of the triple-biomarker（AFP， AFP-L3%， DCP）determinations thus to maximize the clinical efficacies and help improving HCC high-risk surveillance，early diagnosis and prognosis.

Role of serum exosomal miR-23b-3p and miR-4429 in patients with hepatocellular carcinoma

CHEN Wenju, ZHOU Yong, XU Jiajia, WANG Pan

2023, 38(7):  624-628.  DOI: 10.3969/j.issn.1673-8640.2023.07.002

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Objective To investigate the roles of serum exosomal microRNA（miR）-23b-3p and miR-4429 expression levels in diagnosing hepatocellular carcinoma（HCC）. Methods Totally，51 HCC patients，45 chronic hepatitis B（CHB） patients and 40 healthy subjects from Taizhou Central Hospital were enrolled from November 2019 to October 2022. The exosomes in serum were extracted by size exclusion chromatography. Identification was performed by western blot. The expression levels of miR-23b-3p and miR-4429 in the exosomes were determined by real-time fluorescence quantitative polymerase chain reaction. Alpha-fetoprotein（AFP） was determined as well. Receiver operating characteristic（ROC）curve was used to evaluate the diagnostic value of miR-23b-3p，miR-4429 and AFP single and combined determinations in HCC. Results The relative expression levels of serum exosomal miR-23b-3p and miR-4429 and AFP levels in HCC group were higher than those in healthy control group and CHB group（P<0.05）. Serum AFP level in CHB group was higher than that in healthy control group（P<0.05），and there was no statistical significance in the relative expression levels of serum exosomal miR-23b-3p and miR-4429 between the 2 groups（P>0.05）. ROC curve analysis results showed that the areas under curves（AUC） of serum exosomal miR-23b-3p，miR-4429，AFP for the diagnosis of HCC were 0.911，0.876 and 0.857，respectively，and the AUC of combined determination was 0.989. The sensitivity and specificity were 92.2% and 98.0%，respectively. Conclusions Serum exosomal miR-23b-3p and miR-4429 can be used as potential biomarkers for the diagnosis of HCC，and the combined determination is helpful to improve the diagnostic efficiency.

Relationship between soluble growth stimulating gene 2 protein and clinicopathological features and prognosis in patients with colorectal cancer

ZHU Rixiang, YIN Jie

2023, 38(7):  629-633.  DOI: 10.3969/j.issn.1673-8640.2023.07.003

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Objective To investigate the relationship between soluble growth stimulating gene 2（sST2） protein and clinicopathological features and prognosis in patients with colorectal cancer. Methods A total of 101 patients diagnosed with colorectal cancer in Hai'an Hospital of Nantong University from May 2017 to May 2020 were enrolled，and serum sST2 level was determined. Clinical data and clinical laboratory determination results were collected. The optimal cut-off value of sST2 for patient prognosis（death） was evaluated by receiver operating characteristic（ROC）curve. Patient survival state was evaluated by Kaplan-Meier survival curve and Log-rank χ² test. Risk factors affecting overall survival（OS）were assessed using Cox proportional hazard regression analysis. Results The optimal cut-off value for sST2 was 459 pg/mL. The patients were classified into high sST2 group（>459 pg/mL）and low sST2 group（≤459 pg/mL）. Compared with low sST2 group，age，the proportion of Eastern Cooperative Oncology Group（ECOG） score ≥1 and the proportion of TNM stage Ⅲ in high sST2 group were increased（P<0.05），and the hemoglobin（Hb） level and tumor differentiation degree were decreased（P<0.05）. Kaplan-Meier survival curve indicated that the OS of high sST2 group was lower than that of low sST2 group（ Log-rank χ²=27.01，P<0.001）. Multivariate Cox proportional hazard regression analysis showed that ECOG score，TNM stage Ⅱ，TNM stage Ⅲ，moderate to poor tumor differentiation and sST2>459 pg/mL were associated with short OS [hazard ratios（HR） were 1.99，2.32，4.09，1.39，1.59，95% confidence interval（CI）1.23-3.01，1.64-4.32，2.32-8.32，1.03-2.00，1.21-2.08，P<0.05]. Conclusions High sST2 level promotes the progression of colorectal cancer and serves as an independent risk factor for poor prognosis. It can be potentially applied for the evaluation of patient prognosis.

Relationship between PFTK1 expression level and microvessel density and postoperative prognosis in non-small cell lung cancer

CHEN Ying, LI Wei, LIN Tao, YANG Li

2023, 38(7):  634-639.  DOI: 10.3969/j.issn.1673-8640.2023.07.004

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Objective To investigate the relationship between the expression of PFTAIRE protein kinase 1（PFTK1） and microvessel density（MVD） and postoperative prognosis in non-small cell lung cancer（NSCLC） patients. Methods A total of 88 NSCLC patients who underwent surgical treatment at Henan Nanyang Central Hospital from January 2013 to January 2015 were enrolled. Cancer tissues and adjacent tissues（>2 cm from tumor edges） were collected，and PFTK1 expression and MVD count were determined. Spearman correlation analysis was used to evaluate the correlation between PFTK1 expression and MVD count. Kaplan-Meier survival curve and Log rank χ² test were used to analyze the survival status of NSCLC patients. Multivariate Cox regression analysis was used to evaluate the risk factors for prognosis（death） in NSCLC patients. Results Among the 88 NSCLC patients，the total positive rate of PFTK1 was 70.45%（62/88）. There was statistical significance in PFTK1 positive rate and MVD count among NSCLC patients with different TNM stages，tumor diameters and lymph node metastasis（P<0.001，P<0.05），while there was no statistical significance in PFTK1 positive rate and MVD count among NSCLC patients with different sex，ages，pathological classification and histological grading（P>0.05）. MVD count in PFTK1 positive group was higher than that in PFTK1 negative group（P<0.05）. Spearman correlation analysis showed a positive correlation between PFTK1 expression and MVD count（r=0.952，P<0.001）. Multivariate Cox regression analysis showed that TNM stage Ⅲ，lymph node metastasis，tumor diameter >3 cm and PFTK1 positivity were independent risk factors for prognosis（death） in NSCLC patients（P<0.05）. Conclusions PFTK1 is highly expressed in cancer tissues of NSCLC patients，and it is closely related to MVD count and patient prognosis.

Effects of miR-24-3p targeting CHD5 on migration，invasion and radiosensitivity of thyroid cancer cells

PAN Xin, LÜ Ying, LI Yang, LI Rongguo

2023, 38(7):  640-646.  DOI: 10.3969/j.issn.1673-8640.2023.07.005

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Objective To investigate the effects of microRNA（miR）-24-3p regulating the expression of chromodomain helicase DNA-binding protein 5（CHD5） on the migration，invasion and radiosensitivity of thyroid cancer cells. Methods A total of 125 patients with thyroid cancer were enrolled in Xiaoshan First People's Hospital from September 2018 to September 2020. Cancer tissues，paracancer tissues （4 cm from tumor margin） and clinicopathologic data were collected. Human normal thyroid cell line Nthy-ori 3-1 and human thyroid cancer cell lines，KTC1，SW579 and TPC-1，were selected. The expressions of miR-24-3p and CHD5 proteins in cancer tissue，paracancer tissue and 4 kinds of cells were determined. SW579 cells were transfected with miR-24-3p inhibitor anti-miR-24-3p（anti-miR-24-3p group），negative control anti-miR-NC （anti-miR-NC group），miR-24-3p mimic miR-24-3p mimics （miR-24-3p mimics group），negative control miR-NC （miR-NC group），co-transfected anti-miR-24-3p and si-NC （anti-miR-24-3p+si-NC group） ，co-transfected anti-miR-24-3p and si-CHD5 （anti-miR-24-3p+si-CHD5 group）. The migration ability，proliferation ability and radiosensitivity of each group were determined. Dual luciferase reporter assay was used to verify the targeting relationship between miR-24-3p and CHD5. Results The relative expression level of miR-24-3p in cancer tissue and thyroid cancer cell lines（KTC1，SW579 and TPC-1） were higher than those in paracancer tissue and normal thyroid cell line Nthy-ori 3-1（P<0.05）. The relative expression level of CHD5 protein was lower than those in paracancer tissue and normal thyroid cell line Nthy-ori 3-1（P<0.05）. SW579 cells had the highest relative expression of miR-24-3p and the lowest relative expression of CHD5 protein. The expressions of miR-24-3p and CHD5 proteins had statistical significance in thyroid cancer patients with or without lymphatic metastasis and different clinical stages（P<0.001）. The inhibition of miR-24-3p expression could inhibit the migration and invasion abilities of SW579 cells，and the radiosensitivity of SW579 cells was enhanced（P<0.05）. The targeting of miR-24-3p regulated CHD5 expression negatively. Silencing CHD5 could reverse the effects of miR-24-3p expression on migration，invasion and radiosensitivity of SW579 cells（P<0.05）. Conclusions The inhibition of miR-24-3p expression can inhibit the migration and invasion abilities of SW579 cells and enhance their radiosensitivity by up-regulating CHD5.

Correlation of peripheral blood monocyte count and serum anti-phospholipase A2 receptor antibody in patients with nephrotic syndrome

TANG Qingqin, QIAO Longwei, YIN Jingping, ZHANG Sheng, FENG Bin, LIANG Yuting

2023, 38(7):  647-652.  DOI: 10.3969/j.issn.1673-8640.2023.07.006

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Objective To investigate the related factors and clinical role of serum anti-phospholipase A2 receptor antibody（PLA2R-Ab）in patients with nephrotic syndrome. Methods A total of 108 patients for the first time diagnosed with nephrotic syndrome and determined positive for serum PLA2R-Ab in the First Affiliated Hospital of Soochow University from August 2019 to September 2021 were enrolled，and their clinical data and laboratory determination results，including 51 items such as biochemistry，immunity，routine blood test and blood coagulation，were collected. Linear regression analysis was used to analyze the relationship between each index and serum PLA2R-Ab level. According to serum PLA2R-Ab level，the patients were classified into low-level （≤60 RU/mL）group and high-level（>60 RU/mL）group. Logistic regression analysis was used to evaluate the influencing factors of high-level PLA2R-Ab. Results PLA2R-Ab was positively correlated with creatine kinase and fibrinogen（β values were 0.001 and 0.057，respectively），and it was negatively correlated with blood glucose，aspartate aminotransferase，potassium and monocyte count（β values were -0.084，-0.007，-0.247 and -0.662，respectively）. According to the quartiles of monocyte count，all the patients were classified into 4 groups，≤0.35×10⁹/L group（28 cases），（0.36-0.45）×10⁹/L group（26 cases），（0.46-0.55）×10⁹/L group（27 cases） and >0.55×10⁹/L group（27 cases）. There was no statistical significance in serum PLA2R-Ab levels among the 4 groups（P>0.05）. After adjusting confounding factors，such as creatine kinase，blood glucose，aspartate aminotransferase，fibrinogen and potassium，compared with monocyte count ≤0.35×10⁹/L group，lgPLA2R-Ab levels in the other 3 groups were decreased by 0.190，0.117 and 0.281（β values were -0.190，-0.117 and -0.281，P=0.008），and serum lgPLA2R-Ab levels were decreased by 1.55，1.31 and 1.91 RU/mL，respectively. Creatine kinase could increase the risk of high-level PLA2R-Ab [odds ratio（OR）=1.012，95% confidence interval（CI）1.003-1.021，P=0.007]，while monocyte count could decrease the risk of serum high-level PLA2R-Ab in patients with nephrotic syndrome（OR=0.047，95%CI 0.004-0.602，P=0.019）. Conclusions The correlation between peripheral blood monocyte count and serum PLA2R-Ab suggests that peripheral blood monocyte may be involved in the pathogenesis of nephrotic syndrome，especially membranous nephropathy.

BISAP score combined with serum sRAGE for predicting hyperlipidemic acute pancreatitis severity and short-term prognosis

WANG Haiping, LI He

2023, 38(7):  653-658.  DOI: 10.3969/j.issn.1673-8640.2023.07.007

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Objective To investigate the efficacy of bedside index for severity in acute pancreatitis（BISAP） score combined with serum soluble receptor for advanced glycation end product（sRAGE） for predicting the severity and prognosis in hyperlipidemic acute pancreatitis（HLAP）. Methods A total of 142 HLAP patients admitted to the Second Affiliated Hospital of Anhui Medical University from March 2017 to August 2022 were enrolled. The acute physiology and chronic health evaluation Ⅱ（APACHE Ⅱ），sequential organ failure assessment（SOFA） and BISAP were scored. The clinicopathologic data and clinical laboratory determination results were collected，and serum sRAGE levels were determined. The patients were classified into severe HLAP（severe group，54 cases） and non-severe HLAP（non-severe group，88 cases） according to BISAP score. According to the 30 d survival status after admission，the patients were classified into death group（42 cases） and survival group（100 cases）. Logistic regression analysis was used to evaluate the risk factors for 30 d admission death in HLAP patients. Receiver operating characteristic（ROC） curve was used to evaluate the differential diagnosis of severe HLAP and death at 30 d after admission. Results The levels of APACHEⅡ score，SOFA score and sRAGE were increased in severe group compared with non-severe group（P<0.05）. Compared with survival group，C-reactive protein（CRP），sRAGE，BISAP score≥3 points，APACHEⅡ score and SOFA score in death group were increased（P<0.05）. Multivariate Logistic regression analysis showed that increased APACHE Ⅱ score and increased sRAGE level were risk factors for severe HLAP [odds ratios（OR） were 1.17 and 1.01，95% confidence intervals（CI） were 1.03-1.34 and 1.00-1.03，respectively]. BISAP score ≥3 and serum sRAGE level were risk factors for death at 30 d after admission in HLAP patients（OR=1.99 and 1.01，95% CI 1.53-2.98 and 1.00-1.01，respectively）. The area under curve（AUC） of sRAGE for differential diagnosis of severe HLAP was 0.73. The AUC of BISAP score，sRAGE single and combined determinations for HLAP patients were 0.81，0.67 and 0.86，respectively. The AUC of CRP，SOFA score，APACHE Ⅱ score and sRAGE in the differential diagnosis of severe HLAP was 0.77. The AUC of combined determination of CRP，SOFA score，APACHEⅡ score，sRAGE and BISAP score for death in 30 d among HLAP patients after admission was 0.89. Conclusions The sRAGE plays a role in assessing HLAP severity. The combined determination of sRAGE and BISAP score can improve the assessment efficacy of short-term prognosis in HLAP.

Correlation between blood lipid levels and cardiac function indexes in patients with coronary heart disease

MA Jianguo, LI Hongchun

2023, 38(7):  659-664.  DOI: 10.3969/j.issn.1673-8640.2023.07.008

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Objective To investigate the correlation between blood lipid levels and cardiac function indexes in patients with coronary heart disease（CHD）. Methods A total of 200 CHD patients（CHD group） and 200 healthy subjects（control group） in Jiangsu Shuyang Zhongxing Hospital from January 2021 to January 2022 were enrolled. Serum total cholesterol（TC），triglyceride（TG），low-density lipoprotein cholesterol（LDL-C），high-density lipoprotein cholesterol（HDL-C），small and dense low-density lipoprotein cholesterol（sd-LDL-C），apolipoprotein（apo）A and apo B，apo E，lipoprotein（a） [Lp（a）] and lipoprotein-associated phospholipase A2（Lp-PLA2） levels were determined. Cardiac function indexes [end-diastolic volume（EDV），end-systolic volume（ESV），left ventricular ejection fraction（LVEF）] were determined. Pearson correlation analysis，multiple linear correlation analysis and Logistic regression analysis were used to evaluate the relationship between blood lipid levels and cardiac function indexes. Results The levels of apo B，apo E，Lp（a），Lp-PLA2，LDL-C，TG and TC levels and EDV and ESV in CHD group were higher than those in control group（P<0.001），and serum HDL-C and apo A levels and LVEF were lower than those in control group（P<0.001）. There was no statistical significance in serum sd-LDL-C levels between the 2 groups（P>0.05）. Pearson correlation analysis showed that apo B，apo E，Lp（a），Lp-PLA2，LDL-C，TG and TC were positively correlated with EDV and ESV（P<0.001），and they were negatively correlated with LVEF（P<0.001）. HDL-C and apo A were negatively correlated with EDV and ESV（P<0.001），and they were positively correlated with LVEF（P<0.05）. EDV and ESV increased by 0.130 and 0.110 mm for each 1 mmol/L increase of LDL-C（β=0.130 and 0.110）. EDV and ESV increased by 0.220 and 0.189 mm for each 1 mmol/L increase of TC（β= 0.220 and 0.189）. For each 1 mmol/L increase of TC，LVEF decreased by 0.310%（β=-0.310）. EDV and ESV of LDL-C>3.40 mmol/L group and TC>5.17 mmol/L group were higher than those of LDL-C≤3.40 mmol/L group and TC≤5.17 mmol/L group，respectively（P<0.001），and LVEF was lower than LDL-C≤3.40 mmol/L group and TC≤5.17 mmol/L group，respectively（P<0.001）. Logistic regression analysis showed that after adjusting for age，sex，body mass index and diabetes mellitus history，LDL-C>3.40 mmol/L and TC>5.17 mmol/L were risk factors for increasing EDV and ESV and decreasing LVEF（P<0.001）. Conclusions TC and LDL-C are correlated with cardiac function in patients with CHD. Abnormal lipid metabolism may be involved in the pathological process of heart failure in patients with CHD.

Correlation of UGT1A1 gene polymorphism with pregnancy complications and adverse pregnancy outcomes among gestational hypothyroidism patients in Shenzhen Bao'an area

ZHANG Xuming, YIN Weiming, GAO Jing

2023, 38(7):  665-668.  DOI: 10.3969/j.issn.1673-8640.2023.07.009

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Objective To study the relationship between uridine diphosphate-glucuronosyl-transferase 1A1（UGT1A1） gene polymorphism and pregnancy complications and adverse pregnancy outcomes in patients with gestational hypothyroidism in Shenzhen Bao'an area. Methods Totally，160 patients with gestational hypothyroidism in Shenzhen Bao'an Shiyan People's Hospital from April 2019 to April 2021 were enrolled as study group. Another 160 pregnant females with normal thyroid function were enrolled as control group. UGT1A1 gene polymorphism was determined. The pregnancy complications（hypertension，diabetes and intrahepatic cholestasis） and adverse pregnancy outcomes（preterm birth，fetal distress and neonatal asphyxia） in different UGT1A1 genotypes were compared. Results The proportion of UGT1A1 GG genotype in study group was 16.25%，which was lower than that in control group（51.25%）（P<0.001），while the proportions of GA and AA genotypes were 42.50% and 41.25%，which were higher than those in control group（23.75% and 25.00%，respectively）（P<0.001，P<0.05）. In study group，the incidence rates of hypertension，diabetes and intrahepatic cholestasis in patients with AA genotype were 18.18%，21.21% and 12.12%，respectively，which were higher than those with GG genotype（0.00%，3.85% and 0.00%，respectively） and GA genotype（2.94%，4.41% and 0.00%，respectively）（P<0.05）. In study group，the total incidence rate of adverse pregnancy outcomes in AA genotype was 21.21%，which was higher than those in GG genotype（0.00%） and GA genotype（5.88%）（P<0.05）. Conclusions UGT1A1 genotypes of patients with gestational hypothyroidism in Shenzhen Bao'an area are mainly GA and AA，among which AA genotype patients have high risk of pregnancy complications and adverse pregnancy outcomes.

Role of PLR in ankylosing spondylitis patients with positive antinuclear antibody

DUAN Lili, JIANG Chang, ZHOU Dongmei

2023, 38(7):  669-674.  DOI: 10.3969/j.issn.1673-8640.2023.07.010

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Objective To investigate the clinical role of platelet-to-lymphocyte ratio（PLR）in patients with ankylosing spondylitis（AS）combined with positive antinuclear antibody（ANA）. Methods A total of 132 patients with AS in the Affiliated Hospital of Xuzhou Medical University from January 2018 to November 2021 were enrolled. There were 39 cases of ANA positivity and 93 cases of ANA negativity. Totally，98 healthy subjects during the same period were enrolled as control group. The difference of general clinical data between the 2 groups were compared. Spearman correlation analysis was used to evaluate the correlation between indexes. Multivariate Logistic regression analysis was used to investigate the independent risk factors of ANA positivity in AS patients. Receiver operating characteristic（ROC）curve was used to evaluate the risk predictive value of each index for AS combined with ANA positivity. The index difference of ANA positive and negative patients after treatment with adalimumab was analyzed. Results The differences of lymphocytes，platelets，monocytes，red blood cell distribution width（RDW），PLR，monocyte-to-lymphocyte ratio（MLR），neutrophil-to-lymphocyte ratio（NLR），immunoglobulin（Ig） G，IgA，IgM between AS group and healthy control group were statistically significant（P<0.05）. RDW was positively correlated with erythrocyte sedimentation rate（ESR）（r=0.189，P=0.034）. PLR，MLR and NLR were positively correlated with ESR and C-reactive protein（CRP）（P<0.05）. Mean platelet volume（MPV） was negatively correlated with ESR and CRP（r=-0.382 and -0.385，P<0.001）. There was statistical significance in age，ESR，CRP，PLR，IgG，lymphocyte count and disease course between ANA positive and negative groups（P<0.05）. For ANA positive patients，PLR was positively correlated with ESR and CRP（r=0.579 and 0.691，P<0.001）. PLR and disease course are independent risk factors for AS combined with ANA positivity（P<0.05）. The adalimumab treatment effect of patients with positive and negative ANA was followed up，and there was statistical significance in RDW，PLR，ESR and CRP between the 2 groups（P<0.05）. Conclusions The ANA positive patients with AS may have high disease activity and poor treatment effect. PLR may play a clinical guiding role in AS patients with ANA positivity.

Correlation of preoperative RPR in peripheral blood with the prognosis of endometrial carcinoma patients

GUO Jie, LI Haixia, LI Xiaoyun

2023, 38(7):  675-679.  DOI: 10.3969/j.issn.1673-8640.2023.07.011

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Objective To investigate the relationship between preoperative peripheral blood red blood cell distribution width-to-platelet count ratio（RPR） and clinicopathological features and prognosis of patients with endometrial carcinoma（EC）. Methods A retrospective analysis was conducted from September 2013 to October 2016 in the First People's Hospital of Shanzhou District. The clinical data of 123 EC patients（including 5-year follow-up results）were collected. Receiver operating characteristic（ROC） curve was used to evaluate the efficacy of RPR in diagnosing EC. Taking 0.193 as the optimal cut-off value，123 EC patients were classified into high RPR group（45 cases） and low RPR group（78 cases），and the clinicopathological features and prognosis of the 2 groups were compared. Cox regression analysis was used to investigate the prognostic factors of EC patients. Results The proportion of non-adenocarcinoma，maximum tumor diameter ≥2 cm，high the Federation International of Gynecology and Obstetrics（FIGO） stage，lymphovascular space infiltration，high differentiation，lymph metastasis，low pathological grade and recurrence cases in high RPR group were higher than those in low RPR group（P<0.05），and the 5-year overall survival rate（40.0%，18/45） was lower than that in low RPR group（73.1%，57/78）（P<0.05）. The 5-year overall survival rate of EC patients with non-adenocarcinoma，maximum tumor diameter ≥2 cm，FIGO stage Ⅲ，lymphovascular space infiltration，high differentiation，lymph metastasis，recurrence and high RPR was decreased（P<0.05）. Cox regression analysis showed that non-adenocarcinoma，recurrence，FIGO stage Ⅲ and high RPR were independent prognostic risk factors for EC patients（P<0.05）. Conclusions RPR is related to clinicopathological progression in EC patients，and high RPR is an independent prognostic risk factor in EC patients.

Evaluation of 7 oscillating mixing methods for SARS-CoV-2 nucleic acid determination

DU Jinghui, HE Xin, LIU Jing, LIU Ruiyan, BAO Zhijun, WANG Jun, ZHAO Yan, LEI Yaorong, LIU Min, LIU Xu

2023, 38(7):  680-685.  DOI: 10.3969/j.issn.1673-8640.2023.07.012

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Objective To evaluate the effect of 7 oscillating mixing methods for severe acute respiratory syndrome corona virus 2（SARS-CoV-2） nucleic acid determination and determine the optimal parameters of effective method. Methods In the first stage，30 samples（small sample） of each oscillating mixing method were used to evaluate effectiveness and determine the optimal time parameters from morphology，flow cytometry and real-time fluorescence polymerase chain reaction（PCR）. In the second stage，large sample was used to verify the effectiveness of multi-tube vortex treatment and ultrasonic shock under the optimal parameters，and the oscillation effects of multi-tube vortex treatment and ultrasonic waves were compared. Results Morphology and flow cytometry results showed that epithelial cells in preservation solution were increased by 6 oscillating mixing methods，except ultrasound（P<0.05）. Real-time fluorescence PCR showed that there was no decrease in internal reference cycle threshold（Ct） in nucleic acid amplification after reverse and cyclotron shock treatments（P>0.05）. There was no decrease in internal reference Ct in nucleic acid amplification after blowing for 6 times and reciprocating shock for 5 min. The internal reference Ct in nucleic acid amplification after single/multi-tube vortex treatment and ultrasonic shock was decreased（P<0.000 1）. Real-time fluorescence PCR results of large sample further verified the effectiveness of multi-tube vortex treatment for 2 min and ultrasonic shock for 5 min（P<0.000 1）. Conclusions For the pre-treatment of SARS-CoV-2 nucleic acid samples，2 min vortex oscillating mixing method can be adopted，which is simple and easy to use and has good oscillation effect.

Expressions and roles of lncRNA CRNDE and miR-384 in peripheral blood of patients with acute leukemia

WANG Tengfei, CHEN Shan, AN Hebing, ZHOU Lei

2023, 38(7):  686-691.  DOI: 10.3969/j.issn.1673-8640.2023.07.013

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Objective To determine the relative expression levels of long non-coding RNA（lncRNA）colorectal neoplasia differentially expressed gene（CRNDE）and microRNA-384（miR-384）in peripheral blood of patients with acute leukemia，and to investigate the clinical value. Methods From March 2017 to April 2019，106 patients with acute myeloid leukemia（AML） in the Second Hospital of Hebei North College were enrolled as AML group. Totally，100 healthy subjects were enrolled as control group. The relative expression levels of lncRNA CRNDE and miR-384 in serum of AML group and control group were determined by real-time fluorescence quantitative polymerase chain reaction（PCR）. Pearson correlation was used to analyze the correlation between serum lncRNA CRNDE and miR-384 relative expression levels in AML patients. Kaplan-Meier survival curve was used to analyze the relationship between lncRNA CRNDE，miR-384 and the prognosis of AML patients. Cox regression analysis was used to analyze the risk factors affecting the prognosis of AML patients. Results The relative expression level of serum lncRNA CRNDE in AML group was higher than that in control group（P<0.001），and the relative expression level of miR-384 was lower than that in control group（P<0.001）. They were correlated with white blood cell（WBC）count，platelet（PLT） count，the proportion of bone marrow blasts，the National Comprehensive Cancer Network（NCCN） risk stratification and extramedullary infiltration（P<0.05）. The relative expression levels of serum lncRNA CRNDE and miR-384 in AML patients were negatively correlated（r=-0.635，P<0.001）. The overall survival rate and event-free survival rate of lncRNA CRNDE low expression（<1.72） patients were higher than those of lncRNA CRNDE high expression（≥1.72） patients（χ²=27.316 and 18.056，P<0.001），and the overall survival rate and event-free survival rate of high miR-384 expression（≥0.65）patients were higher than those of low miR-384 expression（<0.65） patients（χ²=8.454，P=0.004；χ²=5.333，P=0.021）. Multivariate Cox regression analysis showed that high expression of lncRNA CRNDE and low expression of miR-384 were risk factors for the poor prognosis of AML patients（P=0.003）. Conclusions The expression level of serum lncRNA CRNDE in AML patients is increased，and the expression level of miR-384 is decreased，both of which are related to the pathological characteristics and prognosis of patients. They may be biomarkers for evaluating the prognosis of patients with AML.

Research progress in influenza viruses

CHEN Danyang, ZHENG Siyu, ZHENG Ruilin, SU Jingyao, ZHU Bing, LI Yinghua

2023, 38(7):  696-703.  DOI: 10.3969/j.issn.1673-8640.2023.07.015

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Influenza is a highly infectious acute respiratory disease caused by influenza virus infection. It has severe pathogenicity and high mortality. In the recent years，the study of influenza viruses has made certain progress，but in the development of drugs and vaccines for influenza viruses it is still inadequate，and the development trend of influenza virus variation situation at present can not be predicted，so this needs scientific research workers focusing on the etiology and pathogenic mechanism of influenza viruses. In this review，the etiology，epidemiology，pathogenic mechanism，influenza-associated pneumonia and acute respiratory distress syndrome were reviewed.