Objective To detect the expression of human telomerase reverse transcriptase(hTERT)and silent information regulatory factor 6(Sirt6)in serum of pregnant women with preeclampsia,and explore the value of hTERT and Sirt6 levels in the evaluation of disease severity and pregnancy outcome.Methods A total of 300 patients with preeclampsia who were treated in Shaanxi Provincial People's Hospital from January 2018 to December 2022 were selected as the preeclampsia group,and all pregnant women met the diagnostic criteria for preeclampsia in the Guidelines for the Diagnosis and Treatment of Hypertensive Disorders in Pregnancy(2015).Meanwhile,300 healthy pregnant women who underwent pregnancy examinations in Shaanxi Provincial People's Hospital during the same period were selected as the control group.Preeclampsia group was divided into mild preeclampsia group(n=180)and severe preeclampsia group(n=120)according to the severity of the disease.The preeclampsia group was divided into normal pregnancy group(n=165)and adverse pregnancy group(n=135)according to the occurrence of adverse pregnancy outcomes.Serum hTERT and Sirt6 levels were detected by enzyme-linked immunosorbent assay(ELISA).Spearman correlation analysis was applied to analyze the correlation between serum hTERT and Sirt6 levels and the severity of preeclampsia in patients.Receiver operating characteristic(ROC)curve was applied to evaluate the value of serum hTERT and Sirt6 levels in the diagnosis of preeclampsia and prediction of pregnancy outcomes.Results Compared with the control group serum levels of hTERT(22.15±5.82 ng/ml vs 30.12±9.56 ng/ml)and Sirt6(5.26±1.62 ng/ml vs 7.06±2.29 ng/ml)in preeclampsia group were decreased,and the differences were significant(t=12.334,11.114,all P＜0.001).Compared with the mild preeclampsia group,the serum levels of hTERT(18.28±4.11 ng/ml vs 24.73±6.96 ng/ml)and Sirt6(4.03±1.17 ng/ml vs 6.08±1.92 ng/ml)in the severe preeclampsia group were decreased,and the differences were significant(t=9.142,10.469,all P＜0.001).Compared with the normal pregnancy group,the serum levels of hTERT(17.75±4.61 ng/ml vs 25.75±6.81 ng/ml)and Sirt6(4.06±0.96 ng/ml vs 6.24±2.16 ng/ml)of preeclampsia pregnant women in the adverse pregnancy group were decreased,and the differences were significant(t=11.639,10.878,all P＜0.001).Spearman correlation analysis showed that the levels of hTERT and Sirt6 in serum were negatively correlated with the severity of preeclampsia in patients(r=-0.562,-0.604,all P＜0.001).ROC curve analysis results showed that the area under the curve(95％confidence interval)[AUC(95％CI)]of serum hTERT and Sirt6 in the diagnosis of preeclampsia were 0.711(0.673～0.747)and 0.727(0.689～0.762),respectively.The AUC(95％CI)of the combined diagnosis of preeclampsia was 0.788(0.753～0.820),which was higher than that of the combined diagnosis of preeclampsia(Z=2.719,2.154,P=0.007,0.031).The AUC of serum hTERT and Sirt6 for predicting adverse pregnancy outcomes of preeclampsia were 0.786(0.735～0.831)and 0.783(0.732～0.829),respectively.The AUC(95％CI)of serum HTERT and Sirt6 for predicting adverse pregnancy outcomes of preeclampsia was 0.849(0.804～0.888).It was higher than predicted by the two alone(Z=1.855,1.861,P=0.032,0.031).Conclusion The serum levels of hTERT and Sirt6 in pregnant women with preeclampsia were low,and they were negatively correlated with the disease severity of preeclampsia patients.They may have certain evaluation values for pregnancy outcomes.

Objective By population survey,to explore the epidemiological characteristics of gastric precancerous lesions in Lishui District of Nanjing and provide objective basis for the prevention and treatment of early gastric cancer.Methods From July 2021 to December 2022,21 977 patients who received endoscopy and/or 13C-UBT in Lishui District People's Hospital and 6 medical community units in Nanjing City were retrospectively analyzed for demography characteristics,detection rate of gastric precancerous lesions,and H.Pylori infection rate.Results(1)590 cases of gastric precancerous lesions were detected(detection rate 2.68%);(2)The total detection rate of precancerous lesions and three pathological types in males were all higher than those in females(all P<0.001);(3)The minimum age for the total detection rate of precancerous lesions in males and the mini-mum age for each pathological type were lower than in females(P<0.001,0.009,0.005,0.002);(4)The popu-lation total H.pylori infection rate was 23.10%,the H.pylori infection rate in patients with precancerous lesions was higher than that in non-precancerous lesions(P<0.001),both H.pylori infection rate of male and female in precancerous lesions were all higher than those of non-precancerous lesions of the same sex(all P<0.001),in addition,the H.pylori infection rate of male whether in precancerous or non-precancerous lesions was higher than that of female(all P<0.001);(5)The precancerous lesions detection rate in male,female,and the overall age range of 20～29 to 70～79 years is positively correlated with age growth(P<0.001),and rapidly decreases after the age of 79,the of H.pylori infection rate was also positively correlated with age growth(P<0.001),and the trend of age change(P<0.001)was parallel to the precancerous lesions detection rate.Conclusions The detec-tion rate of gastric precancerous lesions in this region is above the average level in China;the total H.pylori infec-tion rate is at a relatively low level in China;the H.pylori infection rate is parallel to the age trend of the detection rate of gastric precancerous lesions,and increases with age.

Objective:To analyze the biological characteristics, phylogeny and the taxonomic status of strain 7712 (=CGMCC 1.17031=NBRC 113783=KCTC 15766) isolated from a clinical blood sample.Methods:Strain 7712 was identified by the cultural properties, cellular and colonial morphology, physiological and biochemical reactions, matrix-assisted laser desorption ionization time-of-flight mass spectrometry system, and genome correlation index analysis. The genomic phylogenetic tree was construct to analyze the taxonomic position. The virulence factors and resistance genes of strain 7712 and related strains were then compared by the online virulence factor database and online comprehensive antibiotic research database respectively.Results:Strain 7712 was urease negative, gram-negative nonfermenters, which was identified as Ochrobactrum anthropi by VITEK GN card. The 16S rRNA gene analysis showed that the strain was closely related to the members of genera Ochrobactrum and Brucella. The phylogenetic tree showed that strain 7712 was clustered together with Ochrobactrum teleogrylli LCB8 T and Ochrobactrum haematophilum CCUG 38531 T, along with genus Brucella and other Ochrobactrum species. The genome relatedness indexes analysis showed that the average nucleotide identity between strain 7712 and Ochrobactrum teleogrylli LCB8 T was 98.16%, which was higher than the threshold for prokaryotic species. Genetic prediction showed that strain 7712 carried several virulence-related genes and resistance-related genes, of which the existence of OCH gene might be responsible to the resistance to cephalosporin. Conclusions:A case of human infection caused by Ochrobactrum teleogrylli is identified, which would help promote the understanding of biodiversity of genus Ochrobactrum.

Objective:To investigate Helicobactor pylori (H. pylori) infection status and interfamilial transmission pattern in Zhengzhou area. Methods:A cross-sectional study was conducted from September 2020 to march 2021, among 731 individual from 266 families randomly selected from 9 communities of Zhengzhou area. H. pylori infection status was determined by serum antibody tests, and 13C-urea breath test was performed in the previously eradicated population to clarify the current infection status. The individual and familial infection rate, infection status for couples and children and adolescent were analyzed. Results:Among 731 individuals from 266 families, 397 of them were H. pylori positive. The individual infection rate was 54.31% (397/731); among infected individuals 77.83% (307/397) were infected with type Ⅰ strain, 22.67% (90/397) were infected by type Ⅱ strain. Annual household income ( χ2=0.419, 0.410, 0.213, all P>0.05), smoking history (χ 2=0.071, P>0.05), drinking history ( χ2=0.071, P>0.05), dining place ( χ2=0.009, P>0.05), gastrointestinal symptoms ( χ2=0.047, P>0.05), family history of gastric disease ( χ2=0.069, P>0.05), and history of gastric cancer ( χ2=0.004, P>0.05) had no significant differences between H. pylori-positive and -negative groups, but the infection rate in individuals with higher education level was lower ( χ2=4.449, P<0.05). The infection rate was significantly higher in≥18 age groups compared with<18 age groups ( χ2=6.531, 23.362, 20.671, 24.244, 37.948, 14.597 and 5.170, all P<0.05). The familial H. pylori infection rate was 87.59% (233/266), and in 61 families all member were infected (26.18%, 61/233). The positive rate was 23.08% (6/26) in 50 families with children under 18 years when both parents were infected. Among 231 coupled families, both couples were infected in 78 families (33.76%), one couple was infected in 113 families (48.92%), and both couples were not infected in 40 (17.32%). With the increase of marriage time, the infection rate of both spouses increased significantly ( χ2=7.775, 12.662, 15.487, all P<0.05). Conclusions:The distribution of H. pylori infection presents a family cluster pattern, and intrafamilial infection is an important transmission rout of H. pylori. The type I strain of H. pylori is the dominate strain in this area.

Objective: To analyse the genomic characteristics of Novirus(NoV) GII.4 genotype JN010 strain isolated form Jinan in 2017. Methods: Seven pairs of primers were designed and used to amplify the JN010 genome. Sequence analyses, alignment and phylogenetic trees of ORF1 and ORF2 genes were performed using the software Lasergene7.1 and MEGA5.2. At the same time the major protein VP1 amino acid mutations were analyzed. Results: The 7 516 bp complete genome sequence of JN010 strain was obtained, the most mutation sites were located in P2 subdomain of VP1. Two substitutions I293N and H373N of VP1 were locate neighboring epitope A, and R297H mutation happened within epitope A and the site A that binding with histo-blood group antigens(HBGAs). The JN010 strain was GII.Pe/GII.4 genotype and genetically closest to the strains found in Osaka of Japan(GenBank accession number LC066046) and the strains in Zhongshan city of Guangdong (GenBank accession number KY407064) respectively according to ORF1 and ORF2 gene homologous and phylogenetic analysis. Conclusions The NoV GII.4 variant strain JN010 has occurred mutations in the key site of the epitope A and site A that bind with HBGAs, and maybe affect its antigenicity and interaction with HBGAs.

Objective To detect and analyze the levels of 27 serum cytokines expression in EV71-induced Hand Foot and Mouth Disease(HFMD) in Jinan and revealed their correlations with disease severity based on the platform of liquid chip.Methods 43 serum samples were collected from EV71-infected HFMD patients in Jinan in 2009-2014,including 16 mild cases and 27 severe cases.10 serum specimens from healthy people were also collected as controls.27 serum cytokines were analyzed on Bio-Plex liquid chip platform.Results Compared to healthy controls,22 cytokines expression levels increased in severe groups,including IL-1 ra,IL-2,IL-4,IL-5,IL-6,IL-7,IL-8,IL-9,IL-10,IL-12 (p70),IL-13,IL-15,IL-17,etotaxin,G-CSF,IFN-γ,IP-10,MCP-1,PDGF-BB,MIP-1β,RANTES and TNF-α(P ＜ 0.02).21 cytokines expression levels elevated than healthy controls (P ＜ 0.02).GM-CSF was found decreased levels in both mild and severe groups than in the healthy controls(P ＜ 0.02).The correlation analysis showed that 12 cytokines (IL-1 ra,IL-7,IL-9,IL-10,IL-13,IL-17,etotaxin,IP-10,PDGF-BB,MIP-1β,RANTES and TNF-α) were moderately correlated with the severity of EV71-infected HFMD.GM-CSF was negatively correlated with HFMD.Conclusions Many kinds of serum cytokine changed obviously in the EV71-infected HFMD patients and cytokine levels were closely associated with the severity of HFMD.The analysis of liquid chip for serum cytokines provides an efficiently method.

Objective To observe the expression of forkhead or winged helix transcription 3 (Foxp3) in colon cancer tissue and paracancerous tissue,and detect the level of serum interleukin (IL)-17 in colon cancer patients and healthy human,to explore the changes of regulatory T cell (Treg cell) and Th17 cell in the process of occurrence and development of colon cancer.Methods The expression of Foxp3 in 56 patients with colon cancer and paracancerous tissue and 15 cases with normal colon tissue was measured by immunohistochemical SP method and the level of serum IL-17 was determined by enzyme-linked immunosorbent method.Results The level of serum IL-17 in colon cancer tissue was higher than that in normal colon tissue [(9.1 ± 2.3) ng/L vs.(6.2 ± 1.5) ng/L],and there was significant difference (P =0.007).Foxp3 positive cell number in colon cancer tissue was more than that in normal colon tissue and paracancerous tissue (24.1 ± 6.4 vs.2.7 ± 1.1 and 8.7 ± 2.3),paracancerous tissue was more than normal colon tissue,and there was significant difference (P ＜ 0.01).The level of serum IL-17 in colon cancer tissue with TNM Ⅲ was higher than that in TNM Ⅰ-Ⅱ [(8.5 ± 2.1) ng/L vs.(5.4 ± 0.9) ng/L],Foxp3 was more than that in TNM Ⅰ-Ⅱ (25.8 ± 6.2 vs.18.2 ± 4.4),and there was significant difference (P＜ 0.01).There was no significant difference in the level of serum IL-17 between middle-high differentiated adenocarcinoma and low differentiated adenocarcinoma [(9.4 ± 1.1) ng/L vs.(8.9 ± 1.8) ng/L] (P ＞ 0.05).Foxp3 in middlehigh differentiated adenocarcinoma was more than that in low differentiated adenocarcinoma (26.8 ± 5.5 vs.17.2 ± 3.2),and there was significant difference (P ＜ 0.01).Conclusions Detection of IL-17 and Foxp3 can provide a new way of targeting therapy for colon cancer.IL-17 levels and Foxp3 expression are closely related to the immune status of local tumor tissues,the joint detection is benefitial to the further understanding of the patients with tumor immune state,provide basic information for tumor immunotherapy.

Objective To observe the prevalence of anionic trypsinogen (PRSS2) gene G191R mutation in patients with acute pancreatitis (AP) and chronic pancreatitis (CP),and to investigate the effect of PRSS2 gene G191R mutation on susceptibility to pancreatitis.Methods The blood samples of 82 patients with acute pancreatitis,73 patients with chronic pancreatitis and 138 healthy subjects were collected,and genomic DNA was extracted.Nest PCR were performed to amplify PRSS2 gene and restriction fragment length polymorphism (RFLP) was followed by using Hpy188Ⅲ to distinguish the G191R mutation.DNA sequencing analysis was performed to confirm the mutation status.Results The size of nest PCR products was 436 bp.RFLP2 produced 309 bp and 127 bp fragments,which were resulted from PRSS2 gene G191R mutation (GGA →AGA).DNA sequencing analysis of the PCR products further confirmed the PRSS2 gene G191R mutation.Five of eighty-two(6.1％) patients with acute pancreatitis had PRSS2 gene G191R mutation (OR=0.682,95％ CI 0.231 ～ 2.010); one of seventy-three (1.4％) patients with chronic pancreatitis had the mutation (OR =0.145,95％ CI 0.019 ～ 1.145),and the corresponding value in healthy group was 8.7％ (12/138).The G191R mutation rate in patients with chronic pancreatitis was significantly lower than that in healthy group (x2 =0.432,P =0.035),but the G191R mutation rates were not significantly different between AP group and healthy group (x2 =0.487,P =0.485).Conclusions PRSS2 gene G191R mutation facilitates the degradation of anionic trypsin,and may reduce the incidence of chronic pancreatitis.

Objective To compare the genotypes distribution of human papillomavirus (HPV ) infection in tissues of cervical cancers and cervical intraepithelial neoplasias (CIN ) and its clinical significance .Methods The polymerase chain reaction (PCR) and the gene-chips technique were utilized for the detection of 23 kinds of HPV genotypes in the tissue specimens from 192 cases of cervical intraepithelial neoplasia (CIN) and 85 cases of cervical cancers .And the related data of all subjects were analyzed .Results In 192 cases of CIN ,the total positive rate of HPV was 82 .29% (158/192) ,the positive rate of single genotype infection was 46 .88% (90/192) and the positive rate of multiple genotypes infection was 35 .42% (68/192);In 85 cases of cervical cancers ,the to-tal infection rate of HPV was 88 .24% (75/85) ,the positive rate of single genotype infection was 65 .88% (56/85) and the positive rate of multiple genotypes infection was 22 .35% (19/85) .Conclusion PCR combined with the gene-chips technique can be used in the detection of the tissue samples of cervical lesions ,once detection can detect 23 kinds of HPV genotypes with high sensitivity and strong specificity ,which has very important significance to the prevention and treatment of cervical cancer and precancerous lesions and the their vaccine research .

ObjectiveTo examine whether DNA extracted from free edge fingernails specimens from patient after hematopoietic stem cell transplantation (allo-HSCT) could be used for short tandem repeat (STR) genotyping and chimerism analyzing,and to observe the chimerism status in fingernails after allo-HSCT.MethodsPeripheral blood,bone marrow,oral mucosa and free edge fingernail specimens were collected from 25 patients which allo-HSCT were performed in Beijing Dao-pei Hospital during Jul.2009 to Sep.2011 and their donor.Genomic DNA was extracted and 15 STR loci genotyping and chimerism analysis were performed.For the first group which including 12 patients,pairs of fingernail and oral mucosa specimens were collected within one month after allo-HSCT and were comparative analyzed.For the second group which including 13 patients,chimerism status in fingernail samples were analyzed 3 months or longer after allo-HSCT,and 3 patients underwent repeated testing at different times.ResultsFor the first group,4 oral mucosa specimens showed donor chimerism with varying degrees,but no donor chimerism was detected.in all of 12 fingernail specimens.For the second group,6.7％ to 82.6％ donor chimerism was detected in fingernail specimens in 5 out of 13 patients.For the 3 patients underwent repeated testing,donor chimerism was continued negative in one cases,but continued positive in the other 2 cases.ConclusionsFree edge fingernail samples of patients within one month after allo-HSCT can be used for STR typing and chimerism analysis,and it is better than oral mucosa samples.There are cells in allo-HSCT donor graft can differentiate into skin cells,donor derived skin cells chimerism can be formed and persist in some patients.Med,2012,35:23-26)