Table of Content

30 June 2013, Volume 28 Issue 6

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Research on interleukin-18 gene promoter polymorphisms and genetic susceptibility of nasopharyngeal carcinoma

PAN Guogang，LUO Bin，TENG Yuanji，LIANG Lina.

2013, 28(6):  457-461.  DOI: 10.3969/j.issn.1673-8640.2013.06.001

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Objective　To investigate the relationship of the single nucleotide polymorphisms and haplotypes of interleukin-18（IL-18）gene promoter with genetic susceptibility of nasopharyngeal carcinoma（NPC）. Methods　The single nucleotide polymorphisms of IL-18 gene promoter -137G/C（rs 187238 G/C）and -607C/A（rs 1946518 C/A）in 190 patients with NPC and 200 healthy controls were determined by polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）. The frequency of haplotypes and linkage disequilibrium of IL-18 gene in different groups were analyzed. Results　The distributions of IL-18 gene -607C/A polymorphism had no statistical significance between NPC and control groups（P＞0.05），but the distributions of IL-18 gene -137G/C polymorphism were significantly different between them（P＜0.05）. The relative risk suffered from NPC group found that of C allele was 1.730 times of the G allele [odds ratio（OR）＝1.730，95%confidence interval（CI）: 1.227-2.439）]. Being consistent with the results of the genotyping analysis，IL-18 gene -137G/C and -607C/A single nucleotide polymorphisms showed strong linkage disequilibrium，and the frequency of the -137C/-607A haplotype in NPC group was significantly higher than that in control group（P＜0.05）. The -137C/-607A haplotype was associated with a significantly increased risk of NPC（OR＝1.699，95%CI：1.174-2.457）. Conclusions　IL-18 gene -137G/C polymorphism and its -137C/-607A haplotype are associated with NPC，and -137C allele is an important genetic susceptibility gene for NPC.

Investigation on the diagnosis significance of G6PD activity in screening of thalassemia by ROC curve evaluation

YUAN Mingsheng 1，YI Wangyun 2，YE Guojuan 2.

2013, 28(6):  462-465.  DOI: 10.3969/j.issn.1673-8640.2013.06.002

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  Objective　To evaluate the application significance of glucose-6-phosphate dehydrogenase（G6PD）activity detection in screening of thalassemia. Methods　A total of 229 blood samples of pregnant women were detected by polymerase chain reaction（PCR）and reverse dot-blotting as golden standard. They were classified into 2 groups according to genetic testing results: thalassemia group（163 cases）and control group（66 cases）. The G6PD activities were detected，and the results were compared with those of PCR. The sensitivity，specificity，positive predictive value（PPV），negative predictive value（NPV），positive likelihood ratio（+LR）and negative likelihood ratio（-LR）were analyzed. The results of G6PD activity detection in screening of thalassemia were analyzed and evaluated by receiver operating characteristic（ROC）curve. Results　G6PD activity in thalassemia group was（13.16±3.23）U/gHb，which was higher than that in control group[（10.63±2.18）U/gHb]（t＝5.83，P＝0.000）. The area under the ROC curve was 0.757. The optimal cut-off value was 11.95U/gHb. The sensitivity，specificity，PPV，NPV，+LR and -LR were 0.693，0.773，0.88，0.50，3.05 and 2.52，respectively. Conclusions　G6PD activity detection has some screening significance in the diagnosis of thalassemia. The increasing of G6PD activity should be highly suspected as thalassemia to further confirm by gene detection.

Preliminary evaluation of the prognostic value of combining eGFR and Hb in CKD patients with coronary artery disease after percutaneous coronary artery intervention

TANG Hongxia，TANG Xin，JIA Kegang.

2013, 28(6):  466-470.  DOI: 10.3969/j.issn.1673-8640.2013.06.003

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Objective　To evaluate the prognostic value of combining estimated glomerular filtration rate（eGFR）and hemoglobin（Hb）in chronic kidney disaese（CKD）patients with coronary artery disease after percutaneous coronary artery intervention（PCI）in hospital and 4 years after hospitalization. Methods　According to the eGFR and Hb levels，1 005 PCI patients with coronary artery disease were classified into 4 groups:18 cases in eGFR＜60 mL/min and anemia group，52 cases in eGFR＜60 mL/min and normal Hb group，37 cases in eGFR≥60 mL/min and anemia group and 898 cases in eGFR≥60 mL/min and normal Hb group.The levels of serum creatinine and Hb of PCI patients were measured，and the basic situations of sex，age，smoking，body mass index（BMI），hyperlipaemia，hypertension，diabetes mellitus，the presence or absence of prior myocardial infarction and perioperative PCI were compared among the 4 groups. All patients were followed up from 30 d to 4 years，and the median was 3 years.The survival analysis based on COX proportional hazard models.Kaplan-Meier survival analysis was used for comparing survival curves among the 4 groups.The Log-rank statistic was used for comparing survival rate among the groups. Results　Sex，BMI and perioperative PCI had no statistical significance among the 4 groups（P＞0.05）. Smoking，hyperlipaemia，diabetes mellitus，the presence or absence of prior myocardial infarction，counts of narrow coronary artery，age，hypertension and left ventricular ejection fraction（LVEF）were detected with statistical significance among the 4 groups（P＜0.05）. The days after PCI had statistical significance（H＝18.399，P＝0.000）. Through multivariate COX regression correcting BMI，smoking，hypertension，hyperlipaemia，diabetes mellitus，the presence or absence of prior myocardial infarction，counts of narrow coronary artery and perioperative PCI，the relative risk（RR）of the increasing counts of narrow coronary artery with cardiac events was 1.333[95％confidence interval（CI）: 1.023-1.738，P＝0.033]. The RR of perioperative PCI was 3.142（95％CI: 1.974-5.001，P＝0.000）.The RR of Hb was 1.006（95％CI: 0.990-1.023，P＝0.446）.The survival rates in the first，second，third and fourth year were different.The each year survival rates of eGFR＜60 mL/min and anemia group were 94.4%，88.9%，83.3% and 83.3%，of eGFR＜60 mL/min and normal Hb group were 98.1%，94.2%，88.5% and 86.5%，of eGFR≥60mL/min and anemia group were 97.3%，94.6%，91.9% and 91.9%，and of eGFR≥60 mL/min and normal Hb group were 99.7%，97.2%，93.7% and 91.6%.The survival rates had statistical significance among the 4 groups（H＝10.440，P＝0.015），and the survival rate of group with eGFR＜60 mL/min and anemia was significantly lower than those of the other 3 groups. Conclusions　It has a value to use combining eGFR and Hb to evaluate the prognosis of CKD patients with coronary artery disease after PCI who have a high morbidity of major adverse cardiac events.

The clinical significance of serum cystatin C in the diagnosis of severe preeclampsia

YU Miao，ZHANG Li，QIN Bing.

2013, 28(6):  471-474.  DOI: 10.3969/j.issn.1673-8640.2013.06.004

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Objective　To investigate the clinical significance of serum cystatin C（Cys C）in the diagnosis of severe preeclampsia. Methods　The serum samples were collected from 797 normal pregnant women in the different stages of gestation following perinatal healthy examination（normal pregnant group），95 pregnant women with severe preeclampsia and 216 healthy non-pregnant women（healthy non-pregnant group）.The serum Cys C and creatinine（Cr）were measured，and the clinical significance of serum Cys C and Cr in the diagnosis of severe preeclampsia was analyzed by receiver operating characteristic（ROC）curve. Results　The serum levels of Cys C elevated from middle-trimester（P＜0.05），and increased significantly after 28 weeks′ gestation compared to healthy non-pregnant group（P＜0.01）.Compared to normal pregnant group in the same gestational period，all serum levels of Cys C increased in early onset and late onset of severe preeclampsia（P＜0.01）.For the early onset of severe preeclampsia，serum level of Cys C was significantly higher than that for the late onset of severe preeclampsia. However，the levels of Cr increased slightly in the early and late onsets of severe preeclampsia（P＞0.05）. ROC analysis on the levels of Cys C showed that an area under the ROC curve（AUC）was 0.96. Conclusions　The serum level of Cys C reflects really the renal damage of pregnant women for severe preeclampsia，and it has a high diagnosis significance.

Predictive significance of parathyroid hormone determination in patients with hypocalcemia after total thyroidectomy

WANG Tiantian1，SHAO Shan2，PENG Zhenxing1，XU Xianfa1

2013, 28(6):  475-478.  DOI: 10.3969/j.issn.1673-8640.2013.06.005

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Objective　To investigate the early predictive significance of serum 1 h intact parathyroid hormone（1h-iPTH）determination in patients with hypocalcemia after total thyroidectomy. Methods　A total of 103 patients undergoing total thyroidectomy were enrolled，and 1h-iPTH and serum 24 h corrected calcium（24h-cCa）levels were determined after total thyroidectomy 1 h and 24 h. Results　Among the 103 patients undergoing total thyroidectomy，the incidence of postoperative hypocalcemia was 37.9%（39/103），and the 1h-iPTH level in patients with hypocalcemia was lower than that in patients with normocalcemia（P＜0.01）. The incidence of hypocalcemia in patients with 1h-iPTH ＜16 pg/mL was higher than that in patients with 1 h-iPTH≥16 pg/mL（P＜0.01）. When 1h-iPTH＜16 pg/mL was as a predictor for hypocalcemia，the sensitivity，specificity and accuracy were 82.1%，90.6% and 87.4%，respectively. Conclusions　1h-iPTH after total thyroidectomy shows a strong correlation with postoperative hypocalcemia，and could be as an indicator for diagnosing hypocalcemia.

Changes and their significance on peripheral blood T lymphocyte subpopulations and hepatitis B virus specific cytotoxic T lymphocytes in patients with severe hepatitis B

PENG Saijiao 1，LI Weilin 1，ZHANG Jin 2.

2013, 28(6):  479-483.  DOI: 10.3969/j.issn.1673-8640.2013.06.006

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Objective　To investigate the changes of peripheral blood T lymphocyte subpopulations and hepatitis B virus（HBV）specific cytotoxic T lymphocytes（CTL）in patients with severe hepatitis B，and to understand the significance of cellular immunity in the progress of severe hepatitis B.  Methods　By flow cytometry，42 patients with severe hepatitis B（8 cases of death and 34 cases of survival），20 healthy controls，20 patients with acute hepatitis B and 40 patients with chronic hepatitis B were determined for T lymphocyte subpopulations and 3 types of CD8+ CTL levels. The T lymphocyte percentages in severe hepatitis B group were compared with those in healthy control group，severe hepatitis B survival group and death group.  Results　Compared with the other groups，the percentages of CD3+ and CD4+，CD4+/CD8+ ratio and the specific CTL levels of core18-27，polymerase 575-583 and envelope335-343 were significantly lower in chronic severe hepatitis B group（P＜0.05）. CD8+ percentage increased significantly. Compared with severe hepatitis B survival group，the changes of T lymphocyte percentages in severe hepatitis B death group were more obvious.  Conclusions　Cellular immunity function in patients with chronic severe hepatitis B has a sharp decline，lacking of HBV specific CTL response，and the cellular immunity involves in the progress of severe hepatitis.

The application of pregnancy test strip for the simultaneous detection of blood and urine HCG in the diagnosis of woman acute abdomen

LI Chaojin，WU Yurong，CHEN Yanjie，ZHANG Li.

2013, 28(6):  484-486.  DOI: 10.3969/j.issn.1673-8640.2013.06.007

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Objective　To evaluate the application significance of pregnancy test strip for the simultaneous detection of serum and urine human chorionic gonadotrophin（HCG）in the diagnosis of ectopic pregnancy（EP）in woman acute abdomen.  Methods　A total of 362 cases diagnosed as EP in patients with woman acute abdomen and 308 non-pregnant patients with woman acute abdomen were enrolled. Their serum and urine samples were collected. The samples were detected for serum and urine HCG by pregnancy test strip. The performance results of the single detection of serum or urine HCG and the simultaneous detection of serum and urine HCG were compared and analyzed.  Results　In woman acute abdomen，the sensitivity of the simultaneous detection of serum and urine HCG was 100%，which was obviously higher than that of the single detection of urine HCG（73.2%，P＜0.01），and there was no statistical significance with that of the single detection of serum HCG（P＞0.05）. When the results for serum and urine HCG were all positive，the positive predictive value for the diagnosis of EP was 100%. When the results for serum and urine HCG were all negative，the negative predictive value for the diagnosis of EP was 100%.  Conclusions　In the differential diagnosis of woman acute abdomen，the simultaneous detection of serum and urine HCG by pregnancy test strip could improve the detection rate of EP，and could make up for the insufficiency of single detection of serum or urine HCG effectively. It has convenience and reasonable price，and is suitable for the hospitals without the quantitation detection of serum HCG.

Influence of seminal oxidative stress on sperm DNA integrity and other parameters in males with infertility

JIAO Ruibao1，FENG Hengxiao2，TANG Jibin1，FANG Jun3，FENG Junrong4，ZHAN Sanhua5，ZHOU Ping1，CHEN Ran1.

2013, 28(6):  487-491.  DOI: 10.3969/j.issn.1673-8640.2013.06.008

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Objective　To investigate the sperm dynamic parameter，shape parameter and function parameter of seminal oxidative stress in males with infertility.  Methods　Sperm parameters in 81 males with infertility were detected，and they were classified into oligospermia group（sperm concentration＜15×106/mL，5 cases），weak sperm group [progressive rate（PR）＜32%，31 cases，Group Ⅰ]，sperm prior to the movement group（32%＜PR＜50%，25 cases，Group Ⅱ，dynamic forward PR group（PR＞50%，11 cases，Group Ⅲ）and leucocyte sperm disease group（referred as white essence disease group，sperm white blood cell count ＞1×106/mL，9 cases）. A total of 21 healthy subjects were as healthy controls. The semen，sperm concentration and motility rate，PR，sperm membrane function，sperm shape normality，malonaldehyde（MDA），total antioxidant capacity（TAC）and sperm DNA integrity parameters were determined.  Results　In comparison with healthy control group，modified MDA concentration of each group was significantly higher than that in healthy control group（P＜0.05 or P＜0.01），and with the decline of sperm motility，sperm MDA increased. The MDA concentrations in Group Ⅰ，Ⅱ and Ⅲ had statistical significance with those in oligospermia group and leucoyte sperm disease group（P＜0.01）. Semen indices of oxidative stress with sperm concentration，total number of sperm，MDA content and sperm DNA fragmentation index（DFI）had positive correlation，and those with sperm motility rate，PR，hypoosmotic swelling rate（HOS），fast linear motion concentration，fast linear motion number，path velocity（VAP），linear velocity（VSL），TAC and sperm shape normality had negative correlation.  Conclusions　Seminal oxidative stress can cause the changes of sperm membrane function，dynamic parameter，shape parameter and sperm nuclear DNA damage，and may be one of the mechanisms causing male infertility.

The significance of coagulation and fibrinolysis related parameters in the third trimester for predicting the postpartum hemorrhage among low-risk pregnant women

LIU Xiaohua，CHENG Weiwei.

2013, 28(6):  492-495.  DOI: 10.3969/j.issn.1673-8640.2013.06.009

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Objective　To investigate the significance of coagulation and fibrinolysis related parameters in the third trimester for predicting the postpartum hemorrhage among low-risk pregnant women. ?Methods　A total of 2 562 singleton uncomplicated primiparous women were enrolled，and their peripheral blood was taken during 32-33 gestational weeks. The platelet（PLT）count，hemoglobin（Hb），fibrinogen（Fbg）and D-dimer（D-D）levels were determined. These women were followed up till delivery. The 48 of 2 562 women developed postpartum hemorrhage，including 16 of 48 women had severe postpartum hemorrhage. ?Results　The Fbg level in severe postpartum hemorrhage group[（4.3±1.2）g/L] was lower than that in the normal pregnant group[（4.8±0.8）g/L，P＝0.007]. When the participants were divided into 2 groups（normal pregnant group and postpartum hemorrhage group），the postpartum hemorrhage group had higher D-D level than the normal pregnant group[（1.8±1.3）g/L and（1.3±0.9）g/L，P＝0.001]. When the participants were divided into 3 groups（normal pregnant group，postpartum hemorrhage group and severe postpartum hemorrhage group），the D-D level in the severe postpartum hemorrhage group was the highest with statistical significance[（2.4±1.7）g/L and（1.5±0.8）g/L，P＝0.000]；[（2.4±1.7）g/L and（1.3±0.9）g/L，P＝0.000]. The PLT count in severe postpartum hemorrhage group was less than that in normal pregnant group[（172.4±38.9）×109 and（197.8±50.9）×109，P＝0.044）. The Fbg level determined in the third trimester had significance to predict postpartum hemorrhage among low-risk pregnant women [odd ratio（OR）＝ 1.89，95% confidence interval（CI）: 1.68-2.03]. The D-D level had the prediction significance（OR＝2.09，95%CI: 1.78-2.30）. The decreasing PLT count in the third trimester had no obvious prediction significance（OR＝1.50，95%CI:0.68-1.93）. ?Conclusions　The D-D and Fbg levels in the third trimester could predict postpartum hemorrhage.

The correlation analysis between urine conductivity by UF-1000i urine analyzer and urine osmotic pressure

WU Peng 1，WENG Aihan 2.

2013, 28(6):  496-498.  DOI: 10.3969/j.issn.1673-8640.2013.06.010

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Objective　To investigate the correlation between urine conductivity and urine freezing-point osmotic pressure determined by UF-1000i urine analyzer.  Methods　（1）The conductivity levels of urine samples of 1 064 healthy subjects were determined. The reference range of urine conductivity in healthy subjects was obtained.（2）A total of 137 random samples were determined for the urine conductivity by UF-1000i and classified into normal group and normal reference range exceeding group，and the urine freezing-point osmotic pressure was determined by Model 3300 Micro-Osmometer. The results of correlation were analyzed.  Results　（1）The reference range of urine conductivity was 5.56-30.29 ms/cm.（2）The conductivity of normal reference range group was correlated to the osmotic pressure（r＝0.942）. The linear regression equation of osmotic pressure against conductivity was Y＝34.18X－42.29. The conductivity of normal reference range exceeding group was correlated to the osmotic pressure（r＝0.979）.The linear regression equation of osmotic pressure against conductivity was Y＝32.34X＋4.12.  Conclusions　It is more convenient to determine the urine conductivity by UF-1000i together with routine urinary examination in daily work than testing the urine osmotic pressure，which is beneficial for estimating renal concentration and dilution function properly.

The clinical significance of sputum smear Gram staining microscopy

CHEN Xianfeng，ZHOU Tingyin.

2013, 28(6):  499-502.  DOI: 10.3969/j.issn.1673-8640.2013.06.011

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Objective　To compare the results of sputum smear Gram staining microscopy with their culture results in 492 cases of sputum specimens in order to evaluate the clinical significance of sputum smear Gram staining microscopy.  Methods　A total of 492 sputum specimens were collected，and their characters and colors of sputum specimens were observed visually. The sputum smear Gram staining microscopy was performed. According to the results，the sputum specimens were classified into A，B and C categories，and were cultured for bacterial identification. The results of microscopy were compared with the detection rate of bacteria.  Results　There were 368 qualified sputum smear specimens in 492 sputum specimens. The qualified rate was 74.80%. There were 124 substandard specimens，accounting for 25.20%，and 426 bacteria were detected. The bacterial detection rate was 77.72% in the 368 qualified sputum smear specimens，of which 286 specimens detected 368 bacteria. The coincidence rate of the qualified sputum smear results and culture results was 71.19%. The bacterial detection rate was 35.48% in the 124 qualified sputum smear specimens，of which 44 specimens detected 58 bacteria. According to the visual observation，the qualified rates and bacterial detection rates of the mucus，bloody and purulent sputum specimens were relatively high，and that of the water or foam-like sputum specimen was low. The qualified rates and bacterial detection rates of the colorless sputum specimens were low in sputum specimens with different colors.  Conclusions　The sputum smear Gram staining microscopy is one of the most simple and cheapest rapid diagnosis methods of the respiratory tract infections. It results faster than culture，and can provide important information for patients in early diagnosis and treatment. The sputum smear Gram staining microscopy can screen out qualified sputum smear specimens and increase the bacterial detection rate of sputum culture. Therefore，sputum smear Gram staining microscopy has clinical significance.

The application significance of serum insulin-like growth factor 1 determination in the early diagnosis of non-small cell lung cancer

SHI Yue 1，ZHANG Jun 2，CHEN Peng 3.

2013, 28(6):  503-505.  DOI: 10.3969/j.issn.1673-8640.2013.05.012

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Objective　To study the sigificance of insulin-like growth factor 1（IGF-1）in the diagnosis of non-small cell lung cancer.  Methods　The serum IGF-1 levels in 90 patients with non-small cell lung cancer and 20 healthy controls were determined by immunoradiometric quantitation assay.  Results　The levels of serum IGF-1 were significantly higher in non-small cell lung cancer group than in healthy control group [（122.6±20.3）ng/mL and（82.7±20.5）ng/mL，P＜0.05]. The levels of IGF-1 in all patients with lymphatic metastasis [（133.2±21.3）ng/mL] were higher than those in patients without lymphatic metastasis [（108.2±15.1）ng/mL，P＜0.05]. The levels of IGF-1 in patients at Phase Ⅲ-Ⅳ[（129.3±20.1）ng/mL] were higher than those at Phase Ⅰ-Ⅱ [（115.2±15.2）ng/mL，P＜0.05]. In non-small cell lung cancer group，the levels of IGF-1 had a significantly positive correlation with carcinoembryonic antigen（CEA）and cytokeratin-19-fragment（CYERA21-1）respectively（r＝0.58 and r＝0.49，P＜0.01）.  Conclusions　High level of serum IGF-1 may have a significant correlation with the pathogenesis，development and metastasis of non-small cell lung cancer，and IGF-1 may have an application significance for the early diagnosis of lung cancer.

Meta analysis on advanced NSCLC with EGFR mutations treated randomly by EGFR inhibitors

ZHANG Rong，CHEN Weiya，GAN Jiemin，ZHAO Hu，GU Zhaoxiang.

2013, 28(6):  506-510.  DOI: 10.3969/j.issn.1673-8640.2013.06.013

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Objective　To use Meta analysis to evaluate the efficacy of epidermal growth factor receptor（EGFR）- tyrosine kinase inhibitors（TKIs）as first-line treatment for advanced non-small cell lung cancer（NSCLC）with EGFR mutations.  Methods　Relevant literatures were searched through PubMed and Embase databases from January 2004 to April 2012. The implement standards for literature-selection were used to ensure the homogeneity of research. Two reviewers evaluated independently the quality of the included studies and extracted the data. Meta analysis was performed by RevMan 5.1 software. Risk ratio（RR）as efficacy index was chosen by overall response rate（ORR），and hazard ratios（HR）were calculated by progression-free survival（PFS）and overall survival（OS）times. The 95% confidence interval（CI）was calculated simultaneously.  Results　Six phase Ⅲrandomly controlled trials were included for a total of 1 030 patients with NSCLS. This Meta analysis showed a statistical significance in ORR（RR＝2.19，95% CI＝1.68-2.15，P＜0.000 01）and PFS（HR＝0.43，95%CI＝0.35-0.52，P＜0.000 01）for patients who received EGFR-TKIs compared with patients who received chemotherapy. No statistical significance was observed in OS（HR＝0.98，95%CI＝0.81-1.18，P＝0.80）.  Conclusions　EGFR-TKIs are effective as first-line treatment for advanced NSCLC. The EGFR mutation testing should be made for advanced NSCLC patients to predict efficacy to first-line therapy with EGFR-TKIs.

The construction of lentivirus-EZH2 3′UTR vector and the screening for its recombinant

ZHAO Weiwei，PENG You，LI Xiaojiao，LU Xiaojia，WANG Yuping，LIU Cuicui，WANG Lulu，FENG Jing.

2013, 28(6):  511-515.  DOI: 10.3969/j.issn.1673-8640.2013.06.014

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Objective　To construct lentiviral vector carrying human EZH2 3′untranslated region（UTR）and maintain its stable integration of recombinant in MCF-7 cells of breast cancer，and to lay the foundation for the further screening of MicroRNAs（miRNAs）.  Methods　The EZH2 3′UTR was extracted and amplified by real-time fluorescence quantitation polymerase chain reaction（RT-PCR），and was constructed into recombinant lentiviral vector，CTX-lentivirus-EZH2 3′UTR vector. The 293T cells were contransfected with the recombinant lentiviral vector together with lentivirus package plasmid to produce lentiviral particles by liposome method. Cracking liquid cracked virus particles，and virus titer was measured according to the expression level of CMV promoter. MCF-7 cells were infected with recombinant lentiviral vector. Puromycin was used to screen the stable MCF-7 cells. The RT-PCR and Western blotting were determined the integration of EZH2 3′UTR into MCF-7 cells.  Results　EZH2 3′UTR was cloned in lentiviral vector. By RT-PCR，the virus titer reached to 4.3×109 copyies/mL. The optimal concentration for screening stable integration of MCF- 7 cells was 0.2 μg/mL. In infected MCF-7 cells，by RT-PCR and Western blotting，the EZH2 3′UTR mRNA was integrated into MCF-7 genome.The difference between the experimental group and the control group was significant（P＜0.01）.  Conclusions　EZH2 3′UTR lentiviral vector has successfully constructed and maintained stable integration in MCF-7 cells of breast cancer.

The significance of TCT combined with HR-HPV DNA PCR determination for the diagnosis of precancerous lesions

CHEN Yantian，CHEN Qinglong，XIONG Yan，SU Xuetang，HUANG Weigang，SHI Sheng，GUO Yihua，CHEN Rongce.

2013, 28(6):  516-518.  DOI: 10.3969/j.issn.1673-8640.2013.06.015

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Objective　To investigate the application and significance of high risk human papilloma virus（HR-HPV）DNA polymerase chain reaction（PCR）combined thinprep cytology test（TCT）determination for the diagnosis of precancerous lesions.  Methods　A total of 540 patients undergoing TCT determination and showing positive atypical squamous cells of undetermined significance（ASCUS）and above were enrolled and performed HR-HPV DNA PCR，and the results were analyzed.  Results　The number of TCT positive combined HPV positive was 259 cases，including ASCUS 155 cases（59.8％），low-grade squamous intraepithelial lesion（LSIL）81 cases（31.3％），high-grade squamous intraepithelial lesion（HSIL）15 cases（5.8％）and squamous cell carcinoma（SCC）8 cases（3.1％）. In the 259 cases，the positive rates of ASCUS，LSIL，HSIL and SCC were 18.1%（28 cases），24.7%（20 cases），66.7%（10 cases）and 100.0%（8 cases），respectively. The sensitivity of the combination determination of the 2 methods was higher than the sensitivities of HR-HPV determination and TCT determination with statistical significance（P＜0.05）.The positive predictive values of HR-HPV determination，TCT determination and the combination determination of the 2 methods were 25.5%，11.1% and 28.2%，respectively，and the negative predictive values were 68.8%，59.2% and 40.8%，respectively.  Conclusions　TCT combined with HR-HPV DNA PCR in the diagnosis of precancerous lesions is an effective method. It is an approach for the diagnosis of early cervical cancer and precancerous lesions.

Comparison of five methods for the detection of methicillin-resistant Staphylococcus aureus

CHEN Biying，MA Chenyun，LI Zhilan，CHEN Peihong，LU Zhicheng，LANG Lizhong.

2013, 28(6):  519-521.  DOI: 10.3969/j.issn.1673-8640.2013.06.016

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Objective　To evaluate the rapid and accurate methods for the detection of methicillin-resistant Staphylococcus aureus（MRSA）.  Methods　A total of 158 isolates of Staphylococcus aureus and 12 isolates of Staphylococcus epidermidis were collected from clinical specimens of Shanghai Seventh People′s Hospital，and were determined by MRSA ID chromogenic culture medium，oxacillin minimum inhibitory concentration（MIC），cefoxitin disk diffusion and penicillin binding protein 2a（PBP2a）latex agglutination for the detection of MRSA. The results were compared with those of fluorescence polymerase chain reaction（PCR）for mecA gene.  Results　The MRSA ID chromogenic culture medium detected 124 isolates，and the sensitivity and specificity were 100.0% and 91.9%. The cefoxitin disk diffusion detected 119 isolates，and the sensitivity and specificity were all 100.0%. The oxacillin MIC detected 117 isolates，and the sensitivity and specificity were 97.5% and 98.4%. The PBP2a latex agglutination detected 117 isolates，and the sensitivity and specificity were 98.3% and 100.0%. When the 4 kinds of methods compared with the PCR，Kappa values were ≥0.75，and had a good consistency.  Conclusions　Cefoxitin disk diffusion for the detection of MRSA has the highest sensitivity and specificity. Compared with conventional methods，MRSA ID chromogenic culture medium for the detection of MRSA shortens the report time for 24h，and it is a fast，simple and convenient method for the detection of MRSA with good specificity.

Analysis on the blind point survey for cell morphology by virtual microscopy in Shanghai，2011

SONG Ying 1，WANG Qing 1，XU Lei 1，HU Xiaobo 2.

2013, 28(6):  522-526.  DOI: 10.3969/j.issn.1673-8640.2013.06.017

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Objective　In order to evaluate the applicability of virtual microscopy of cell morphology，Shanghai Center for Clinical Laboratory had investigated the quality of cell morphology by virtual microscopy in hospitals above Level 2 Grade A and some community hospitals from October to November in 2011.  Methods　A total of 15 cases from the established cell library were selected. The difficult degrees of these cases were similar. These cases were given by the World Health Organization haematology center. The double blind marshaling method was used，and the ability of blood cell morphological recognition of these hospitals were evaluated by scene investigation.  Results　The satisfaction of cell image quality investigation reached 97.5%. The survey satisfaction was 96.3%. For white blood cell count，the results of virtual microscopy and the result given by World Health Organization in counting average value，standard deviation and bias % with the paired t test statistics were P＝0.436 0，P＝0.150 5 and P＝0.180 6. There was no statistical significance（P＞0.05）. However，according to Rümke statistical count ranges with the reference given by World Health Organization，there was bias，and should be corrected. For more than 80% of the inspection items，the cell recognition rate was＞60%，which was better than those of previously used blood smear method. In addition to strengthening the recognition of cell morphology，the establishment of a characteristic cell count classification standard should be considered.  Conclusions　Due to the virtual microscopy with image quality and clear cell distinguishment，the method has the advantages of standardization，quality and reliability，and can be accepted for most cell morphology and is worth popularizing.