Table of Content

30 September 2020, Volume 35 Issue 9

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Mixed infection status of human papillomavirus,Ureaplasma urealyticum,Chlamydia trachomatis and Neisseria gonorrhoeae in Changning district of Shanghai

XUAN Binbin, TAN Meiyu, SUN Hanxiao, SHENG Huiming

2020, 35(9):  859-863.  DOI: 10.3969/j.issn.1673-8640.2020.09.001

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Objective To investigate the mixed infection status and correlations of human papillomavirus（HPV） with Ureaplasma urealyticum（UU）,Chlamydia trachomatis（CT） and Neisseria gonorrhoeae（NG） among female patients in Changning district of Shanghai. Methods Totally,4 496 samples,including cervical exudate cell and cervical swab samples,of female patients from Shanghai Jiao Tong University School of Medicine Tongren Hospital were collected from June 1st 2018 to May 31st 2019. A total of 113 healthy subjects were enrolled as healthy control group. CT DNA,UU DNA and NG DNA were determined by fluorescence probe-polymerase chain reaction（PCR）,and PCR-reverse dot blot was used to determine HPV DNA subtypes. The positive rate and subtype distribution characteristics were analyzed and compared with those of healthy control group. The determination rates of CT DNA,UU DNA and NG DNA were investigated by classification according to the negativity or positivity and infection types of HPV. Results The determination rate of UU DNA was the highest（65.9%）,followed by HPV DNA（23.9%）,and the determination rate of NG DNA was the lowest（0.9%）. Compared with healthy control group,the determination rates of UU DNA,CT DNA and NG DNA were 62.8%（71/113）,3.6%（4/113）and 0.9%（1/113）,and the determination rate of HPV was 22.1%. The subtype with the highest determination rate of HPV DNA was high-risk HPV52. Compared with the other types,the determination rates of CT,NG and UU were also high（23.2%,2.9% and 20.9%,respectively）. The determination rates of UU,CT and NG in HPV positive group were higher than those in HPV negative group（P<0.001）. With the increasing number of HPV DNA subtypes,the positive rates of CT,NG and UU were increased. The positive rates of CT and UU in patients with 4 or more subtypes of HPV infections（20.00% and 87.7%） were higher than those in patients with single subtype of HPV infection（10.8% and 76.9%）（P<0.05）. Conclusions The determination rate of UU is higher than those of HPV,NG and CT. With the occurrence of multiple infection of different subtypes of HPV,the infection rates of CT,UU and NG increase,and mixed infection occurs. It should pay attention to the occurrence of multiple infection of HPV.

Analysis on drug resistance and toxin gene carrying status of Clostridium difficile clinical isolates in a hospital of Beijing

SHAO Donghua, SONG Yan, LIANG Guowei

2020, 35(9):  864-867.  DOI: 10.3969/j.issn.1673-8640.2020.09.002

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Objective To understand the drug resistance and toxin gene carrying status of Clostridium difficile clinical isolates in Aerospace Center Hospital. Methods The stool specimens of clinical unexplained diarrhea patients were detected by TaqMan-MGB probe real-time fluorescence quantitative polymerase chain reaction（PCR） to identify genus gene and toxin gene. Anaerobic culture,isolation and drug susceptibility test were conducted for the positive specimens of tpi gene. The clinical records were analyzed retrospectively,in order to compare the difference between A⁺B⁺ isolates and A^-B⁺ isolates. Results The tpi gene of 56 specimens was positive in 335 stool specimens,the positive rate was 16.7%,in which 10（17.9%） isolates were A^-B⁺ isolates,and 46（82.1%） isolates were A⁺B⁺ isolates. The drug resistance rates to amoxicillin,metronidazole,clindamycin,vancomycin,moxifloxacin,ampicillin-sulbactam and meropenem were 7.1%,10.7%,75.0%,0,57.1%,5.4% and 8.9%,respectively. There were 29（51.8%） isolates with drug resistance to both clindamycin and moxifloxacin. There was no statistical significance in patient history,clinical drug history and laboratory data（P>0.05）. Conclusions The multiple drug resistance of Clostridium difficile in Aerospace Center Hospital is common. The clinical characteristics are not different between A⁺B⁺ isolates and A^-B⁺ isolates.

Distribution and seasonal prevalence of pathogens causing diarrhea in a hospital of Shenzhen

LIANG Cuiqiong, TANG Meiling, XIE Zhihua

2020, 35(9):  868-871.  DOI: 10.3969/j.issn.1673-8640.2020.09.003

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Objective To investigate the distribution and seasonal prevalence of pathogens（rotavirus,norovirus,adenovirus and astrovirus） causing diarrhea in Longhua District Center Hospital of Shenzhen. Methods A total of 1 050 children aged 1 month-5 years with acute diarrhea admitted in Longhua District Center Hospital of Shenzhen were enrolled. The stool samples were collected and determined by enzyme-linked immunosorbent assay（ELISA） for 4 common kinds of intestinal virus（rotavirus,norovirus,adenovirus and astrovirus） antigens,which were confirmed by multiple polymerase chain reaction（PCR） for 4 virus nucleic acids. Results In 1 050 children with acute diarrhea,483 cases of positive virus antigen samples were determined,the determination rate was 46.00%. Totally,595 cases were positive for nucleic acid,and the determination rate was 56.67%. The consistency rate was 75.86%. The determination of nucleic acid positive by multiple PCR was used as the standard. The most common virus was rotavirus（23.24%）,followed by norovirus（17.50%）,adenovirus（9.62%） and astrovirus（6.76%）. Viral infection was mainly in the form of single infection（45.05%）,and mixed infection was less（11.62%）. There was no statistical significance in positive determination rate with different sex（χ²=0.210,P=0.647）,and there was no statistical significance in positive determination rate between children <3 years old and children 3-5 years old（χ²=46.663,P=0.001）. The incidence of rotavirus infection peaked in April,norovirus infection peaked in July,and the incidence rates of adenovirus and astrovirus infections increased in November and December. Conclusions Rotavirus and norovirus are the important viruses in children with viral diarrhea in Longhua District Center Hospital of Shenzhen from 2015 to 2017,and the intestinal infections of the 4 main viruses are characterized by seasonal epidemics.

Correlation between urinary protein and disease progression in COVID-19 patients

ZHANG Tengfei, QI Weiqiang, LING Yun, ZAI Shubei, WANG Yan, FAN Jian, ZHANG Bei, ZHU Zhaoqin

2020, 35(9):  872-874.  DOI: 10.3969/j.issn.1673-8640.2020.09.004

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Objective To investigate the correlation between urinary protein and disease progression in patients with corona virus disease 2019（COVID-19）. Methods The fresh urine samples from 268 COVID-19 patients（246 cases of light/common type and 22 cases of severe/critical type） within 3 d of admission and 155 discharged patients were collected. Dry chemical analysis was used for routine urine test. Totally,125 suspected patients without severe acute respiratory syndrome coronavirus 2（SARS-CoV-2）infection were enrolled as control group. Results The positive rates of urinary protein in severe/critical type group and light/common type group were higher than that in control group at admission （P<0.05）. The positive rate of urinary protein in severe/critical type group was higher than that in light/common type group（P<0.000 1）,and there was no statistical significance for the positive rate between the 2 groups at dismission（P>0.05）. There were 23.53%（20/85） patients in 20-39 years old group with positive urinary protein at admission and negative at dismission. There were 38.20%（34/89）patients in 40-59 years old group with positive urinary protein at admission and 5.26% patients with still positive urinary protein at dismission. Creatinine and urea nitrogen in severe/critical type group were higher than those in light/common type group（P<0.000 1）. Conclusions Urinary protein has an indicative effect on the progress of COVID-19.

Role of ctDNA KRAS mutation determination for therapeutic effectiveness assessment of neo-adjuvant therapy in patients with colorectal cancer

FANG Yongyun, LU Daru, SHEN Lisong

2020, 35(9):  875-880.  DOI: 10.3969/j.issn.1673-8640.2020.09.005

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Objective To investigate the role of circulating tumor DNA （ctDNA） V-Ki-ras2 Kirsten ratsarcoma viral oncogene homolog （KRAS） mutation determination for therapeutic effectiveness assessment of neo-adjuvant therapy in locally advanced colorectal cancer patients （cT3N1-2M0 or cT4N0-2M0）. Methods Totally,53 patients with KRAS-mutated locally advanced colorectal cancer who received neo-adjuvant therapy were enrolled. The ctDNA KRAS mutation mutant-allele frequencies （MAF） before and after neo-adjuvant therapy were determined by droplet digital polymerase chain reaction （ddPCR）. The results of computed tomography （CT） of the chest,abdomen and pelvis were collected,and serum carcinoembryonic antigen （CEA） and carbohydrate antigen 19-9 （CA19-9） were determined. Results The MAF values of ctDNA KRAS mutation before neo-adjuvant therapy had statistical significance compared with that after neo-adjuvant therapy in the 53 locally advanced colorectal cancer patients （P<0.05）. With MAF difference （MAF before treatment–MAF after treatment） as dependent variable,the comparative analysis using least-significant difference （LSD） demonstrated that MAF differences were statistically significant between-in the grading groups classified by therapeutic effectiveness assessment with both radiography and pathology （P<0.05）. The changes of serum CEA and CA19-9 （levels before treatment–levels after treatment） were also statistically significant between-in the grading groups of radiographic and pathological therapeutic effectiveness assessment（P<0.05）. Conclusions Peripheral blood ctDNA KRAS mutation allele frequency can be used as a new molecular marker to evaluate the therapeutic effectiveness of neo-adjuvant therapy,which supplements the traditional therapeutic effectiveness assessment of locally advanced colorectal cancer.

Diagnostic role of combined detection of serum ADA and LDH and the percentage of lymphocyte subsets in peripheral blood in infectious mononucleosis

ZHANG Li, ZHU Hongsheng, MA Jun, JIN Yi, SHEN Donghua

2020, 35(9):  881-884.  DOI: 10.3969/j.issn.1673-8640.2020.09.006

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Objective To investigate the diagnostic role of combined detection of serum adenosine deaminase（ADA）and lactate dehydrogenase（LDH） and CD4⁺/CD8⁺ ratio and CD19⁺B lymphocyte percentage in peripheral blood in infectious mononucleosis（IM）. Methods A total of 78 children with IM were enrolled as experimental group,80 healthy children were enrolled as healthy control group,and 88 children with bronchopneumonia or acute upper respiratory tract infection were enrolled as disease control group. Blood specimens of children from experimental group and disease control group were collected at the first visit. Serum levels of ADA and LDH were determined by peroxidase method and rate method,and the CD4⁺/CD8⁺ ratio and the percentage of CD19⁺B lymphocyte in peripheral blood were determined by flow cytometry. Linear correlation analysis was used to analyze the correlation of serum ADA and LDH levels in IM group and the correlation of peripheral blood CD4⁺/CD8⁺ ratio and CD19⁺B lymphocyte percentage. Receiver operating characteristic（ROC） curve analysis was used,and diagnostic efficiency was evaluated. Results Serum ADA and LDH in experimental group were higher than those in control groups（P<0.01）,and there was a positive correlation between them（r=0.445,P<0.01）. The CD4⁺/CD8⁺ ratio and the percentage of CD19⁺B lymphocyte in experimental group were lower than those in control groups（P<0.01）,and there was a positive correlation between them（r=0.681,P<0.01）. The areas under curves of ADA,LDH,CD4⁺/CD8⁺ ratio and the percentage of CD19⁺B lymphocyte were 0.983,0.911,0.992 and 0.986,respectively. The area under curve of the combined detection was 0.998. Conclusions There are high sensitivity and specificity of the combined detection of serum ADA and LDH and the percentage of peripheral blood lymphocyte subsets in diagnosing IM,and it is beneficial for the early diagnosis of IM.

Correlation and risk assessment of serum 25（OH）D level and multiple thyroid function indicators in patients with thyroid cancer before operation

CHEN Gang, WANG Jing, DENG Qiang, CHEN Xi, HU Dong, ZHUANG Lidong

2020, 35(9):  885-888.  DOI: 10.3969/j.issn.1673-8640.2020.09.007

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Objective To investigate the changes of serum 25-hydroxyvitamin D [25（OH）D] levels in patients with thyroid cancer and the correlation with serum thyroid function indicators and the risk of thyroid cancer. Methods A total of 56 patients with thyroid cancer（thyroid cancer group） and 69 patients with benign thyroid nodules（benign nodule group） were enrolled. Serum 25（OH）D,free triiodothyronine（FT₃） and free thyroxine（FT₄）,thyroid-stimulating hormone（TSH）,thyroglobulin（Tg）,thyroid peroxidase antibody（TPOAb） and thyroglobulin antibody（TgAb） levels were determined. According to the level of 25（OH）D,all the subjects were classified into 3 groups,including vitamin D deficiency（≤20 ng/mL）,vitamin D insufficiency（>20-≤30 ng/mL）and vitamin D sufficiency（>30 ng/mL） groups. Spearman correlation analysis was used to evaluate the correlation among various indicators. Odds ratio （OR） and 95% confidence interval （CI） were used to evaluate the risk of thyroid cancer. Results Serum level of 25（OH）D in thyroid cancer group was lower than that in benign nodule group（P<0.01）,and serum TSH level was higher（P<0.05）. There was no statistical significance in the other indicators between the 2 groups（P>0.05）. In thyroid cancer group,25（OH）D was negatively correlated with TSH and TPOAb（r=-0.750 and -0.872,respectively,P<0.01）,and it had no correlation with FT₃,FT₄,Tg and TgAb levels（r=0.104,0.03,-0.134 and -0.149,respectively,P>0.05）. The risk of thyroid cancer was increased when 25（OH）D≤20 ng/mL （OR=7.990,95% CI 1.640-38.100）. Conclusions 25（OH）D levels are correlated with thyroid cancer,and low 25（OH）D level may increase the risk of thyroid cancer.

Investigation of lipoprotein-associated phospholipase A2 levels among apparently healthy population in Wuhan

LIANG Tao, LIU Feng, ZHANG Detai, WANG Lin, HU Lihua

2020, 35(9):  889-894.  DOI: 10.3969/j.issn.1673-8640.2020.09.008

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Objective To investigate the distribution of lipoprotein-associated phospholipase A2（Lp-PLA2） among apparently healthy population in Wuhan. Methods Totally,1 503 apparently healthy subjects were enrolled. The levels of serum amyloid A（SAA）,Lp-PLA2 and homocysteine（Hcy） were determined. The data fo sex and age and the determination result data of total bilirubin （TB）,direct bilirubin （DBil）,alanine aminotransferase （ALT）,aspartate aminotransferase （AST）,alkaline phosphatase （ALP）,gamma-glutamyltransferase （GGT）,blood urea nitrogen （BUN）,creatinine （Cr）,uric acid （UA）,cystatin C （Cys C）,glucose （Glu）,total cholesterol （TC）,triglyceride （TG）,high-density lipoprotein cholesterol （HDL-C）,low-density lipoprotein cholesterol （LDL-C）,apolipoprotein A （apo A）,apolipoprotein B （apo B） and lipoprotein（a）[Lp（a）] were collected. The correlation between Lp-PLA2 and the other indicators was analyzed by Spearman correlation. Results Lp-PLA2 ≥200 ng/mL was used as increasing（positive）. The Lp-PLA2 level and positive rate of males were higher than those of females（P<0.001）. The positive rate of Lp-PLA2 in ≥61-year-old group was higher than those in 41-60-year-old group and ≤40-year-old group,and serum Lp-PLA2 level was the highest in 41-60-year-old group. In increased Lp-PLA2 group,the age,SAA,ALT,AST,ALP,GGT,BUN,UA,Cr,Cys C,Glu,TG,apo B and Hcy were higher than those in normal Lp-PLA2 group（P<0.05）. TB,DBil,HDL-C and apo A were lower than those in normal Lp-PLA2 group（P<0.05）. There was no statistical significance in TC,LDL-C and Lp（a） between the 2 groups（P>0.05）. The levels of Lp-PLA2,SAA,GGT and Glu in Lp-PLA2 highly increased（>400 ng/mL） group were higher than those in Lp-PLA2 moderately increased（301-400 ng/mL） and slightly increased（200-300 ng/mL） groups（P<0.05）. There was no statistical significance for the other indicators among the 3 groups（P>0.05）. Spearman correlation analysis showed that Lp-PLA2 was positively correlated with SAA and GGT（r=0.422 and 0.162,P<0.001 and P=0.001,respectively）,and it had no correlation with the other indicators（P>0.05）. Conclusions Lp-PLA2 is high among apparently healthy population in Wuhan. Lp-PLA2 screening is necessary in apparently healthy population.

Roles of dynamic monitoring of SARS-CoV-2 antibodies in the diagnosis of COVID-19

HUANG Shaokun, ZHONG Qingyang, ZHU Haipeng, LUO Rundi, ZHANG Jianming

2020, 35(9):  895-899.  DOI: 10.3969/j.issn.1673-8640.2020.09.09

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Objective To investigate the roles of severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） immunoglobulin（Ig） M and IgG antibodies in corona virus disease 2019（COVID-19）. Methods A total of 80 patients with COVID-19 were enrolled,and 20 patients without COVID-19 were enrolled as control group. Immunochromatography was used to determine SARS-CoV-2 IgM and IgG antibodies. A total of 23 samples from COVID-19 patients were collected at different time points for the determination of SARS-CoV-2 antibodies. Results The specificities of SARS-CoV-2 IgM antibody,IgG antibody and IgM+IgG antibodies were all 100%,and the sensitivities were 76.3%,81.3% and 90.0%,respectively. The overall consistency rates were 81.0%,85.0% and 92.0%,respectively. Within 7 d of onset,the sensitivities of SARS-CoV-2 IgM antibody,IgG antibody and IgM+IgG antibodies were 65.0%,70.0% and 70.0%,on 8-14 d of onset,the sensitivities were 72.7 %,95.5% and 95.5%,and after 15 d of onset,the sensitivities were 84.2%,97.4% and 97.4%,respectively. SARS-CoV-2 IgM antibody showed a tendency to become positive on the 5th to 13th d after onset,and the tendency to turn negative was observed after the 18th d of onset. SARS-CoV-2 IgG antibody showed a tendency to turn positive on the 5th d and continued to be positive thereafter. Conclusions Serum SARS-CoV-2 IgM and IgG antibodies can be used as auxiliary diagnostic indicators of COVID-19,and the continuous monitoring of them can help with judging disease process.

Roles of the detections of 8 pathogen IgM antibodies in patients with different pathogens of respiratory tract infection

SHEN Xiaohua, ZHU Lijie

2020, 35(9):  900-902.  DOI: 10.3969/j.issn.1673-8640.2020.09.010

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Objective To investigate the roles of the detections of 8 pathogen IgM antibodies in patients with different pathogens of respiratory tract infection. Methods From June 2018 to June 2019,3 600 patients with respiratory tract infection were enrolled from Jiaxing Hospital of Traditional Chinese Medicine of Zhejiang Chinese Medical University. Indirect fluorescence immunoassay was used to detect 8 pathogen IgM antibodies,including adenovirus（ADV）,Legionella pneumophila（LP）,Chlamydia pneumoniae（CP）,respiratory syncytial virus（RSV）,influenza A virus（IFA）,Mycoplasma pneumoniae（MP）,influenza B virus（IFB） and parainfluenza virus（PIVS） 1,2 and 3. Results A total of 522 cases of pathogen IgM antibodies were detected in 3 600 patients with respiratory tract infection,with a total positive rate of 14.50%. IFB-IgM,IFA-IgM and MP-IgM accounted for 6.11%,4.36% and 2.17%,respectively. There was a statistical significance for the positive rates of IgM antibody detection in MP,IFA,IFB and PIVS in different seasons（P<0.05）. The infection rate of MP in autumn and winter was higher than that in spring and summer,and the infection rates of IFA and IFB in winter and spring were higher than those in summer and autumn. The infection rate of PIVS in spring and summer was higher than that in autumn and winter（P<0.05）. The infection rates of LP,CP,ADV and RSV in different seasons had no statistical significance（P>0.05）. The positive rate of single infection was the highest among all the patients with respiratory tract infection,and the positive rate of infection decreased gradually with the increase of the number of pathogens（P<0.05）. There was no statistical significance in the probability of infection of different pathogens among all the groups（P>0.05）. Conclusions The detection of IgM antibodies of 8 pathogens is of significance for the early diagnosis of respiratory tract infection,and it has guiding significance for the prevention of multiple infections in multiple infection season.

Changes and clinical significance of liver function and oxidative stress in patients with nonalcoholic fatty liver

WANG Yongbin, LU Kuiying, SU Yong, ZHANG Jun, LIU Fudi

2020, 35(9):  903-907.  DOI: 10.3969/j.issn.1673-8640.2020.09.011

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Objective To investigate the changes and clinical significance of liver function and oxidative stress in patients with nonalcoholic fatty liver,and to provide a reference for clinical diagnosis and treatment. Methods A total of 120 patients with nonalcoholic fatty liver were enrolled as study group,and 100 healthy subjects were enrolled as control group. The liver function and oxidative stress were compared between the 2 groups. The correlation of liver function and oxidative stress with nonalcoholic fatty liver was analyzed by Pearson single factor analysis,and the diagnostic value of liver function,oxidative stress and the combined determination in patients with nonalcoholic fatty liver was analyzed by receiver operating characteristic（ROC）curve. Results The levels of alanine aminotransferase（ALT）,aspartate aminotransferase（AST） and gamma-glutamyltransferase（GGT） in study group [（43.69±6.12） U/L,（36.37±5.04） U/L,（48.07±6.41） U/L] were higher than those in control group [（23.36±3.37） U/L,（21.64±3.15） U/L,（20.85±3.08） U/L]（P<0.05）. The levels of glutathione（GSH） and malondialdehyde（MDA） in study group were （321.84±42.75）μmol/L and （6.30±2.02）U/mL,which were higher than those in control group [（270.61±30.37） μmol/L,（2.25±1.40） U/mL]. The levels of catalase（CAT） and total superoxide dismutase（SOD） were （40.72±6.20）U/mL and （80.27±11.78）U/mL,which were lower than those in control group [（53.42±6.91） U/mL,（92.82±12.27） U/mL]（P<0.05）. Pearson correlation analysis showed that liver function,GSH and MDA levels were positively correlated with nonalcoholic fatty liver（r=0.835,0.722 and 0.741,P<0.05）,and CAT and SOD levels were negatively correlated with nonalcoholic fatty liver（r=-0.685 and -0.712,P<0.05）. ROC curve analysis showed that the area under curve（AUC）,sensitivity and specificity of the combined determination were 0.862,86.79% and 93.24%,respectively,which were higher than those of the single determinations of liver function and oxidative stress（0.670,62.03%,81.82%;0.754,77.64%,64.55%）（P<0.05）. Conclusions Nonalcoholic fatty liver patients have severe liver function and oxidative stress damage. The higher the liver function,GSH and MDA levels are,and the lower the CAT and SOD levels are,the greater the risk of nonalcoholic fatty liver is. The combined determination of liver function and oxidative stress can effectively improve the sensitivity and specificity in the diagnosis of nonalcoholic fatty liver,and it is worthy of clinical application.

Correlations of serum adipsin and Vit D levels with islet beta cell function and insulin sensitivity in patients with type 2 diabetes mellitus

ZHANG Qianjin, HU Jin'e, HU Yichuan, DING Ying, SHEN Yanqiu, CHEN Huiqin

2020, 35(9):  908-911.  DOI: 10.3969/j.issn.1673-8640.2020.09.012

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Objective To investigate the correlations of serum adipsin and vitamin D（Vit D） with islet beta cell function and insulin sensitivity in patients with type 2 diabetes mellitus（T2DM）. Methods A total of 60 patients with T2DM and 60 patients with impaired glucose tolerance（IGT） were enrolled,and 60 healthy subjects were enrolled as control group. Serum adipsin,fasting plasma glucose（FPG）,fasting insulin（FINS） and Vit D were determined. The acute insulin response（AIR）,the homeostasis model assessment beta（HOMA-β） and insulin sensitivity index（ISI） were calculated. Multivariate Logistics regression analysis was used to study the influencing factors of serum adipsin and Vit D,and Pearson correlation analysis was used to study the correlations of serum adipsin and Vit D with AIR,HOMA-β and ISI. Results The levels of serum adipsin and Vit D in T2DM group were lower than those in IGT group and control group,while the levels of FPG and FINS were higher than those in IGT group and control group（P<0.05）. Body mass index（BMI）and waist-to-hip ratio had statistical significance among the 3 groups（P<0.05）,and sex and age had no statistical significance（P<0.05）. HOMA-β,ISI and AIR were the risk factors for serum adipsin and Vit D levels（P<0.05）. Serum adipsin and Vit D in T2DM patients were positively correlated with HOMA-β and ISI（P<0.05）,while serum adipsin and Vit D were positively correlated（P<0.05）. Conclusions Serum adipsin and Vit D levels are correlated with islet beta cell function and insulin sensitivity in patients with T2DM.

Changes and roles of sST2,SF and hepcidin levels in patients with CHF and anemia

ZHANG Hongxia, HAN Hong

2020, 35(9):  912-916.  DOI: 10.3969/j.issn.1673-8640.2020.09.013

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Objective To investigate the changes and roles of soluble suppression of tumorigenicity 2（sST2）,serum ferritin（SF） and hepcidin in patients with chronic heart failure（CHF） and anemia. Methods A total of 182 CHF patients were enrolled. According to the presence or absence of anemia,they were classified into CHF-anemia group（89 cases） and CHF-non-anemia group（93 cases）. Totally,43 healthy subjects were enrolled as healthy control group. The clinical data and laboratory determination results [fasting blood glucose（FBG）,C-reactive protein（CRP）,N-terminal pro-B-type natriuretic peptide（NT-proBNP）,hemoglobin（Hb）,hematocrit（HCT） and serum iron] were collected. The levels of sST2,SF and hepcidin were determined. The correlations were evaluated by Pearson correlation analysis. The risk factors of CHF combined with anemia were evaluated by Logistic regression analysis. The diagnostic role of each index for CHF combined with anemia was evaluated by receiver operating characteristic（ROC） curve. Results The levels of CRP,NT-proBNP and sST2 in CHF-anemia group and CHF-non-anemia group were higher than those in healthy control group（P<0.05）. The levels of CRP,NT-proBNP,sST2 and hepcidin in CHF-anemia group were higher than those in CHF-non-anemia group（P<0.05）,while the levels of Hb,serum iron and SF were lower（P<0.05）. Logistic regression analysis showed that the increased levels of CRP,NT-proBNP,sST2 and hepcidin were risk factors for CHF combined with anemia（P<0.05）,while the increased levels of Hb,HCT,serum iron and SF were protective factors（P<0.05）. Pearson correlation analysis showed that Hb and SF were negatively correlated with sST2（r=-0.167 8 and -0.291 4,P<0.05）,while hepcidin was positively correlated with sST2（r=0.353 3,P<0.05）. The results of ROC curve analysis showed that the areas under curves（AUC） of Hb,HCT,sST2,SF and hepcidin for the diagnosis of CHF combined with anemia were 0.640,0.531,0.734,0.789 and 0.845,respectively. The AUC of sST2 combined with SF and hepcidin for the diagnosis of CHF combined with anemia was 0.912. Conclusions sST2,SF and hepcidin are related to CHF combined with anemia,which may be applied as auxiliary diagnostic indexes.

Role of Xpert MTB/RIF system in the rapid detection of Mycobacterium tuberculosis and rifampicin resistance

LIU Jun, LIU Tao, DAI Wanqin, WANG Yixiang, HOU Yanqiang

2020, 35(9):  920-923.  DOI: 10.3969/j.issn.1673-8640.2020.09.015

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Objective To investigate the role of Xpert MTB/RIF system in the rapid detection of Mycobacterium tuberculosis（MTB） and rifampicin resistance. Methods The sputum samples of 1 562 suspected tuberculosis patients were collected,and were determined by Xpert MTB/RIF,liquid culture and acid resistance staining method. The sensitivity and specificity of Xpert MTB/RIF for detecting acid resistance staining positive samples were analyzed using liquid culture method as the standard. The identification and the drug susceptibility test were carried out on 188 cases of positive liquid culture samples to evaluate the sensitivity and specificity of the detection of MTB and rifampicin resistance by Xpert MTB/RIF. Results The consistency rate of Xpert MTB/RIF with liquid culture was 93.92%（χ²=3.55,P>0.05）. That with smear method was 89.44 %（χ²=48.51,P<0.001）. The sensitivity of Xpert MTB/RIF was 82.41%（328/398）,and the specificity was 97.85%（1 139/1 164）,using liquid culture as a standard,with no statistical significance between Xpert MTB/RIF and liquid culture（χ²=3.55,P>0.05）. Totally,188 positive liquid culture samples were confirmed by final identification,the sensitivity of Xpert MTB/RIF was 91.23%（156/171）,the specificity was 88.24%（15/17）,the accuracy was 90.96%（171/188）,and there was no statistical significance between Xpert MTB/RIF and liquid culture（χ²=4.109,P>0.05）. The sensitivity of Xpert MTB/RIF to rifampicin resistance was 82.35%（14/17）,the specificity was 99.35%（1/154）,and there was no statistical significance between Xpert MTB/RIF and traditional proportional method（χ²=0.138,P>0.05）. Conclusions The Xpert MTB/RIF system could detect MTB and rifampicin resistance rapidly,with high sensitivity and specificity,and it is potential in the clinical application.

Influence of dry heat inactive treatment for the determination of inflammatory indicators related with corona virus disease 2019

ZHAO Minghai, LI Xin, WEI Dong, LUO Min, ZHANG Yajie, YANG Yiqi, ZHENG Lei

2020, 35(9):  924-927.  DOI: 10.3969/j.issn.1673-8640.2020.09.016

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Objective To investigate the influence of dry heat inactive treatment（56 ℃ 30 min） for the determination of inflammatory indicators related with corona virus disease 2019（COVID-19）diagnosis and treatment. Methods Totally,332 clinical residual blood samples which C-reactive protein（CRP）,procalcitonin（PCT） and interleukin 6（IL-6） determinations were carried out were collected. All the samples were inactivated by dry heat inactive treatment at 56℃ for 30 min on the day of collection,and the difference of results was compared. Results There was no statistical significance in CRP determination results before and after inactivation（P=0.167）,and there was statistical significance in IL-6 and PCT determination results before and after inactivation（P=0.002,P<0.001）,of which the PCT determination results of 98（79.03%） samples decreased after inactivation. The determination results of the 3 inflammatory indicators before and after inactivation had good consistency,and PCT,CRP and IL-6 had 96.77%,97.11% and 97.11% scatter points in 95% confidence intervals,respectively. The correlation coefficients （r） were 0.992,0.999 and 0.996,respectively. Conclusions Dry heat inactive treatment at 56℃ for 30 min has no effect on CRP determination results,but has effect on PCT and IL-6 determination results. It should pay attention to the pretreatment process of dry heat inactive treatment,in order to reduce the infectious risk of COVID-19 patients' samples.

Prokaryotic expression and serological verification of antigenic epitopes of human papillomavirus type 16 L1 protein

HOU Jianghou, ZHANG Lingxia, SUN Wenna, LIU Yanhua, YANG Bingfen, SUN Weiguo

2020, 35(9):  928-932.  DOI: 10.3969/j.issn.1673-8640.2020.09.017

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Objective To obtain antigenic epitopes of human papillomavirus（HPV） type 16 L1 protein expression and purification in prokaryotic system,and to evaluate its serological reaction. Methods The nucleotide sequence of HPV type 16 L1 protein antigenic epitopes were synthesized and cloned into prokaryotic expression vector pET-DsbC. The constructed recombinant plasmid pET-DsbC-HPV16 L1 was transformed to Escherichia coli BL21 Rosetta（DE3） for expression under the induction of isopropyl-beta-D-thiogalactoside（IPTG）. The L1 protein was expressed and purified by metal chelate affinity chromatography. The antigenicity and immunological activity of fusion protein were evaluated with clinical serum samples by western blotting and enzyme-linked immunosorbent assay（ELISA）. Results The expression vector pET-DsbC-HPV16 L1 was successfully constructed and stably expressed in Escherichia coli Rosetta （DE3）. The recombinant HPV16 L1 protein was purified by affinity chromatography,which had a relative molecular mass of 45 000 and a purity of 95%. The purified rate of recombinant protein was 22%。The results of western blotting showed that the recombinant DsbC-HPV16 L1 protein could produce specific antigen-antibody reaction with HPV16 positive serum,but there was no cross reaction with recombinant DsbC protein or healthy human serum,which had good specificity. ELISA results also indicated that recombinant protein had a higher level of reactivity to serum from patients than serum from healthy or phosphate-buffered saline（PBS）,the A₄₅₀ values were 0.56,0.24 and 0.21,respectively. Conclusions HPV type 16 L1 protein antigentic epitopes are overexpressed solubly in prokaryotic system and have a high immunogenicity and immunological activity.

Analysis of spike protein antigenic epitopes of SARS-CoV-2,HCoV-OC43 and HCoV-HKU1

ZENG Jiantao, LI Quan, ZHOU Lili, DONG Shanshan, ZHENG Zhaobin

2020, 35(9):  933-936.  DOI: 10.3969/j.issn.1673-8640.2020.09.018

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Objective To study the characteristics of spike（S） protein antigenic epitopes of severe acute respiratory syndrome coronavirus 2（SARS-CoV-2）,human coronavirus（HCoV）-OC43 and HCoV-HKU1. Methods The S protein sequences and spatial structures of SARS-CoV-2,HCoV-OC43 and HCoV-HKU1 were obtained from the National Center for Biotechnology Information（NCBI）. Multiple sequence alignment was used to analyze these protein sequences. Protean,IEDB and SYFPEITHI were used to predict epitopes. Results From the 65 complete SARS-CoV-2 S protein sequences included in NCBI,9 sequences of S proteins with no amino acid repeats were selected,and QHZ87592.1 was used as the standard sequence. The other 8 S protein sequences had 8 site mutations and 1 site deletion,but none of them were distributed in receptor binding domain（RBD）. RBD regions of S protein of SARS-CoV-2,HCoV-OC43 and HCoV-HKU1 were different. The amino acid sequences of the S2 subunits of the 3 coronavirus S proteins were highly similar. The B cell antigenic epitopes of S protein were mainly concentrated in S1 subunit,and there was no similarity among the 3 coronaviruses. The T cell antigenic epitopes of the S2 subunit of HCoV-HKU1 and HCoV-OC43 were all similar to that of SARS-CoV-2,and the highest similarity was 70.0%. Conclusions The T cell antigenic epitopes of the S2 subunit may be a common antigen of the 3 coronaviruses. Memory T lymphocytes sensitized by HCoV-OC43 and HCoV-HKU1 might play roles in heterosubtypic immunity to SARS-CoV-2 .

Role of apolipoprotein M in the proliferation,migration and invasion of non-small cell lung cancer A549 cells

WANG Zijin, ZHANG Xiaoying

2020, 35(9):  937-942.  DOI: 10.3969/j.issn.1673-8640.2020.09.019

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Objective To investigate the influence of apolipoprotein M（apo M） on the proliferation,migration and invasion of non-small cell lung cancer（NSCLC） A549 cell line and its relationship with matrix metalloproteinase（MMP）-10. Methods A549 cells being transfected with apo M lentivirus were used as apo M-OE group,and those being transfected with A549 empty lentivirus were used as negative control group. CCK-8 method,scratch test and Transwell invasion test were used to determine the proliferation,migration and invasion abilities. The expression levels of apo M and MMP-10 were determined by real-time fluorescence quantitative polymerase chain reaction（qRT-PCR） and immunoblotting. Results Compared with negative control group,the expression level of apo M in apo M-OE group was up-regulated（P<0.01）,and the expression level was about 28 times higher than that of negative control group. The proliferation and invasion rates of apo M-OE group were higher than those of negative control group（P<0.001）. Compared with negative control group,the blank field of vision in apo M-OE group was reduced,the closing rate was accelerated,and the migration rate was greater（P<0.001）. The expression levels of MMP-10 mRNA,apo M protein and MMP-10 protein in apo M-OE group were higher than those in negative control group（P<0.05）. Conclusions Over-expressing apo M can promote the proliferation,migration and invasion of A549 cells,and can make the cells highly expressing MMP-10.

Application of PDCA cycle in promoting mutual recognition of biochemical item determination results

LIU Zhangling, LI Juan, PU Ran, TANG Rongrui

2020, 35(9):  943-945.  DOI: 10.3969/j.issn.1673-8640.2020.09.020

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Objective To investigate the role of PDCA cycle management model in promoting mutual recognition of biochemical item determination results. Methods The quality control,overall passing rate and qualified rate of fresh blood sample comparison of 31 hospitals in Shapingba District of Chongqing before and after PDCA cycle were compared. Results Totally,18（58.0%） units passed 14 biochemical items（passing rate ≥75%） in laboratory departments of 31 hospitals in 2016,and 28（90.3%） units in 2017 and 31（100%） units in 2018 passed. In 2016,the qualified rate of only 1（7.1%） of the 14 biochemical items was >90%. In 2017 and 2018,the qualified rates were 8（57.1%） and 13（92.9%）,respectively. Conclusions With PDCA cycle management model,the overall passing rate of biochemical items and the qualified rate of sample comparison in clinical laboratories have been improved effectively,which provides a reference for promoting the mutual recognition of determination results.

Plasma ctDNA determination in the diagnosis and treatment of colorectal cancer

CHEN Xinning, WU Shengchao, GUO Wei, WANG Beili

2020, 35(9):  948-951.  DOI: 10.3969/j.issn.1673-8640.2020.09.022

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The incidence and mortality of colorectal cancer are increasing. It is because of the lack of suitable means for early screening,and the compliance of colonoscopy among patients is not ideal. Another reason is the limitations of treatment methods,and some patients' lesions can not be resected. There are uncertainties in the efficiency of chemotherapy and radiotherapy,and targeted therapy is only available for certain population. Through molecular diagnostic techniques,plasma circulating tumor DNA（ctDNA） can be determined,and early screening,prognosis and chemotherapeutic drug response can be performed,which can guide clinical treatment. The review focuses on the determination of plasma ctDNA and its application in the diagnosis and treatment of colorectal cancer.

Research progress of circulating tumor DNA in renal cell carcinoma

SHI Yue, DONG Dong

2020, 35(9):  952-956.  DOI: 10.3969/j.issn.1673-8640.2020.09.023

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Renal cell carcinoma（RCC） is the 7th most common cancer in the world. Due to the lack of symptoms in early stage,many patients are diagnosed with advanced disease and with poor prognosis. Circulating tumor DNA（ctDNA） is tumor-derived fragment DNA in blood or body fluid,which can reflect entire tumor genome and is easy to obtain. It is potential in tumor diagnosis,prognosis assessment,disease recurrence monitoring,therapeutic effect monitoring and chemotherapy resistance monitoring. Recent studies have shown that ctDNA is also clinically valuable in RCC. This review describes the recent progress in the application of ctDNA in the clinical diagnosis and treatment of RCC.