Objective To establish human choriocarcinoma JeG-3 cell line resistant to floxuridine (FUDR)and describe the characteristics of this FUDR-resistant subline.The thymidylate synthase (TS) expression level in FUDR-resistant subline was also discussed.Methods The FUDR-resistant sub-line JeG-3/FUDRA was established by intermitted exposure to grads increased FUDR.Reversed microscope was used to observe the morphological changes in FUDR-resistant sub-line.Population doubling time was calculated and compared based on the growth curve of these two cell lines,cell cycles and chromosomal ploidy were assayed with flow cytometry methods.The chemo-luminescence assay was used to detect the hormone secretion by two kinds of cell lines.The resistant index (RI) was measured by cell counting kit-8 (CCK-8)assay.Quantitative RT-PCR was used to detect the mRNA expression level of TS and we also detected the TS mRNA expression level in different doses exposed subline.Results The RI of JeG-3/FUDRA was 31.62.Compared with the JeG-3 cell,the FUDR-resistant cell line had gross changes in morphological,cell growth,cell cycles and chromosomal numbers.The ability of human chorionic gonadotrop(hCG) and progesterone secretion was lower in JeG-3/FUDRA subline.The trend of TS mRNA expression was:while exposed to low concentration of FUDR,the TS mRNA expression level was downregulated,then followed the increasing dose of the drug,the expression level of TS mRNA ascended gradually.When the terminal concentration was reached,the expression level of TS mRNA in JeG-3/FUDRA subline was higher than that of JeG-3 cell line (P＜0.05).Conclusions We established the FUDR-resistant subline of JeG-3 successfully.The TS mRNA expression level is stage-related to the different concentration and different phase in FUDR exposure.Our data suggested that TS mRNA expression level may not be used as a biomarker to predict the chemosensitivity in FUDR-based chemotherapy.

Objective To explore the clinical characteristics and medicine treatment of patients with severe ulcerative colitis (UC),the efficacy of rescue treatment in patients with glucocorticoid (GCS) resistant severe UC,and the clinical risk factors in patients with GCS-refractory severe UC.Methods From January 2001 to December 2012,clinical,laboratory,endoscopy,imaging data and medication of treatment of 106 patients with severe UC were retrospectively analyzed.Then the patients were followed up,and the clinical efficacy and under endoscopic presentation of Mayo score were evaluated.Logistic regression analysis was performed to analyze the high risk factor of GCS-refractory severe UC.Results Among 106 patients with severe UC,95 were chronic relapse type accounting for 89.6 ％.The percentage of patients with defecation times over six was 73.6％ (78/106),with severe purulent bloody stool was 51.0％ (54/106),and with moderately or severe abdominal pain was 83.0％ (88/106).The percentage of diffuse colon type was 83.0％ (88/106),endoscopic presentation of Mayo score over two was 87.7％ (93/106).Hemoglobin decreased in 65.1％ (69/106) patients,blood platelet increased in 48.1％ (51/106) patients,C-reaction protein elevated in 88.7％ (94/106) patients,and hypoalbuminemia decreased in 42.5％ (45/106) patients.Account to 89.6％ (95/106) of patients with severe UC received GCS treatment,and the percentage of induced remission was 64.2％ (61/95),effective rate was 16.8％ (16/95),and ineffective rate was 18.9％ (18/95).The percentage of GCS refractory was 35.8％(34/95).There were 23 patients with GCS resistance and 11 patients with GCS dependence.Ten patients with GCS resistant severe UC accepted medicine rescue therapy.Five cases were treated with cyclosporin A,of which two cases induced remission,one case was effective,and two cases were ineffective.Another five cases were treated with infliximab,of which three cases induced remission,and two cases were ineffective.The results of Logistic regression analysis showed that severe anemia (OR=6.750,95％CI:2.656 to 17.152,P＜0.01),elevated blood platelets (OR=4.032,95％CI:1.226 to 13.261,P=0.015) and albumin level less than 25 g/L (OR =3.022,95 ％ CI:1.236 to 7.390,P =0.022) were risk factors of GCS-refractory severe UC.Conclusions GCS resistant or dependent occurred in part of patients with severe UC.Patients with G-CS resistant severe UC receive rescue treatment of cyclosporin A or infliximab.Severe anemia,elevated blood platelets,albumin less than 25 g/L may be clinical predicting factors in patients with GCS-refractory severe UC.

Objective To retrospectively analyze the diagnostic value of a noninvasive score system based on transient elastography (TE),serological test and imaging examination on esophageal variceal bleeding (EVB) in patients with hepatitis B virus (HBV)-related cirrhosis.Methods Between April 2011 and December 2012,172 patients with HBV-related cirrhosis including 120 males and 52 females who visited clinic or hospitalized at the Department of Hepatology,Tianjin Third Central Hospital,were retrospectively enrolled.The mean age was (52.9 ± 10.6) years.Patients underwent upper gastrointestinal endoscopy to evaluate esophageal varices (EV) and were further categorized into three stages of mild,moderate and severe according to the morphology of EV and the risk of bleeding.Liver stiffness and spleen stiffness measurement were performed using Fibroscan.Portal vein width,splenic width and spleen thickness were measured using color Doppler ultrasound.All the patients were tested for white blood cell counts and platelet counts.With endoscopy as the gold standard,receiver operating characteristic (ROC) curves and the areas under curves (AUC) were used to assess the performance of the noninvasive score system in predicting EV by liver stiffness,spleen stiffness,portal vein width,spleen thickness and platelet counts.Student's t-test was performed to determine differences between continuous variables.Pearson's correlation was used to evaluate the association between EVB and these parameters.Results All these 172 patients underwent endoscopy.Among them,41 were EVB patients and 131 with no bleeding of EV.Among 172 EV patients,39 without EV,30 were mild EV,47 were moderate EV and 56 were severe EV.EVB was all positively correlated with liver and spleen stiffness,portal vein width,spleen thickness,splenic vein width (r=0.224,0.771,0.214,0.425 and 0.364,respectively; all P＜0.05).EVB was negatively correlated with platelet counts (r=-0.408,P=0.000).Liver stiffness,spleen stiffness,portal vein width,spleen thickness and splenic vein width in EVB patients were significantly higher than those in EV patients (P＜0.05).In contrast,platelet counts level was lower in EVB patients with difference of statistical significance (P＜0.05).AUC of non-invasive score system for EV and EVB were 0.953 and 0.882,respectively (P＜0.05).The optimal cut-off level of noninvasive score system for prediction of EV and EBV were 7 (sensitivity:96 ％,specificity:85 ％) in EV patients and 10 (sensitivity:78％,specificity:89 ％) in EVB patients.Conclusion Non-invasive score system based on liver stiffness,spleen stiffness,spleen thickness,width of splenic and portal vein and platelet counts is of clinical importance in assessing the presence of EV in patients with HBV-related cirrhosis,which is higher clinically valuable in the diagnosis for EV.

Objective To investigate the value of spleen stiffness measured by transient elastography (FibroScan,FS) for diagnosing esophageal-gastric varices in liver cirrhosis patients.Methods A total of 259 cirrhotic patients in Tianjin Third Central Hospital from Apr 2011 to Apr 2012,and 30 healthy controls were enrolled.All the patients and controls were evaluated for spleen and liver stiffness by FS and 201 cirrhotic patients also underwent gastroscopy for the diagnosis of esophageal-gastric varices.By using gastroscopy as the gold standard,the receiver operating characteristic (ROC) curves of three parameters including spleen stiffness,liver stiffness and platelet/thickness of spleen were delineated for different disease stage.The areas under curves (AUC) were used to evaluate the value of these parameters in the diagnosis of esophageal-gastric varices.Results The spleen and liver stiffness values in cirrhotic patients were (44.64 ± 22.27) kPa and (24.27 ±18.89) kPa,respectively,while those in healthy controls were (20.94± 14.78) kPa and (6.12±5.77) kPa,respectively,which were both lower than cirrhotic patients (P＜0.05).The stiffness values of liver and spleen both increased with higher Child-Pugh scores.And the liver stiffness values were different among groups (F=0.068,P =0.000),while the spleen stiffness values in patients with Child-Pugh A and B were different from that in patients with Child-Pugh C (P＜0.05).In patients with moderate or serious esophageal-gastric varices,the spleen and liver stiffness values were significantly higher.The ROC curve analysis showed that the AUC of spleen stiffness,liver stiffness and platelet/thickness of spleen in the patients with moderate to serious esophageal-gastric varices were 0.918,0.749 and 0.743,respectively.The corresponding optimal cut-off values were 50.7 kPa,20.1 kPa and 1.65.The AUC,sensitivity and specificity of spleen stiffness were all higher than liver stiffness and platelet/thickness of spleen.Conclusion Spleen stiffness measured by transient elastography is a valuable parameter for non-invasive diagnosis of esophageal-gastric varices in cirrhotic patients.

Objective: To investigate the effects of novel targeted non-viral vector in gene therapy of human glioma. Methods: The EGF-R targeting gene delivery system GE7 was constructed. Human Glioma cell line U251 was transfected in vitro with ?－gal as reportor gene and p21 as therapeutic gene using this gene delivery system. By means of the assay of ?-galactosidase staining, Western blotting, in situ end labeling apoptosis cells and DNA ladder, the transferring of exogenous genes and the apoptosis of the tumor cells were examined.Results: It was showed that gene transfer efficiency is over 80%. When transfected with p21 gene, the growth of cells was inhibited significantly, and the apoptosis was detected in the transfected cell by the methods of in situ end labeling and DNA ladder. Conclusion: The GE7 gene delivery system has the ability to transfer exogenous gene to tumor cells and the expression of the therapeutic gene can inhibit the growth of the cells.

The purpose of this study is to obtain effective and stable bacteriocin-like substance from lactic acid bacteria. Lactobacillus buchneri KLDS1.0364,which was isolated from fermented cream,a traditional dairy product in Inner Mongolia,could produce bacteriocin-like substance. The bacteriocin-like substance produced by KLDS1.0364 was partially purified and the characteristics were studied. The bacteriocin-like substance was purified by SP-Sepharose fast flow cation exchange chromatography. The molecular weight of the bacteriocin-like substance was 21.6kD,as determined by tricine-SDS-PAGE. The bacteriocin-like substance remained stable after 30 min at 121℃ and after 2 h of incubation during pH 2～10. Complete inactivation of antimicrobial activity was observed after treatment of the bacteriocin-like substance with several proteinases. Treatment with catalase and ?-amylase did not result in any changes of antimicrobial activity. The mode of action of the bacteriocin-like substance is bactericidal. It exhibited a broad spectrum of antagonistic activity against various species of Gram-positive bacteria,Gram-negative bacteria and fungi.

Objective To investigate the age distribution and types of human papillomavirus (HPV) cervical infection in patients in Wuhan region, so as to provide scientific basis for the prevention and treatment of HPV infection and cervical cancer.Methods 9 915 exfoliated cervical cell specimens from patients in Wuhan Medical and Health Center for Women and Children between January 2015 and March 2016 were performed HPV genotyping (21 subtypes), clinical data of 1 732 HPV positive cases were statistically analyzed.Results HPV-positive rates in ≤25, 26~, 36~, 46~, and ≥56 age groups were 22.15%, 15.90%, 17.04%, 19.97%, and 17.57%,respectively (χ2=36.587,P<0.01),HPV-positive rates in ≤25 and 46~ age groups were both higher than other age groups.There were significant differences in single infection and multiple infection rates among different age groups respectively(χ2=14.39, 36.51,respectively, both P<0.05),single infection rate was highest in 46~ and ≤25 age groups (15.41% and 15.24% respectively);multiple infection rates was highest in ≤25 and ≥56 age groups (6.90% and 5.86% respectively).The percentage of single infection and multiple infection were 75.58% and 24.42% respectively, the major single infection type was HPV high-risk subtype (84.34%),the major multiple infection types were high-risk and high-risk compound subtype as well as high-risk and low-risk compound subtype, accounting for 60.52% and 38.77% respectively.The main single infection types were HPV high-risk subtypes 33, 68, 31 and 16, as well as and low-risk subtype 11, the ratios of single infection to multiple infection were 3.13, 2.03, 1.71, 1.67 and 2.00 respectively.Conclusion Cervical infection rates in women in Wuhan region is high in ≤25 and 46~ age groups, there are differences in the distribution of different HPV subtypes of single infection and multiple infection.

This study aimed to investigate viral infections and the prevalence of influenza-like illness (ILI) in Shijiazhuang, China, in 2011 and to provide a scientific basis for the diagnosis and control of respiratory tract infections. Throat swab specimens were collected from 483 cases of ILI who were outpatients in the influenza surveillance sentinel hospitals in Shijiazhuang between January and December 2011. All specimens were examined by multiplex RT-PCR for the following 15 respiratory tract viruses: adenovirus (ADV), human rhinovirus (HRV), human parainfluenza virus (PIV types 1-4), influenza virus A (FluA), influenza virus B (FluB), human enterovirus (HEV), respiratory syncytial virus (RSV-A and -B), human metapneumovirus (HMPV), human coronavirus (HCoV-229E/NL63 and -OC43/HKU1), and human bocavirus (HBoV). Among the 483 cases of ILI, 214 (44.31%) were positive for viruses, including ADV (8.7%), HEV (8.7%), RSV-A (8.07%), HRV (7.45%), FluA (5.38%), HCoV-OC43/ HKU1 (2.9%), PIV-3 (2.9%), HMPV (1.86%), PIV-1 (1.24%), HCoV-229E/NL63 (1.04%), PIV-2 (1.04%), HBoV (0.83%), and FluB (0.41%). Twenty-six (5.38%) of all cases were co-infected with two or more viruses, most commonly HEV/HRV with other viruses. Cases of viral infection were detected throughout the year, with peaks in January and February. ADV and HRV were detected throughout almost the whole year without obvious seasonality. HEV was detected between April and November, with a peak of prevalence in summer and autumn. FluA and FluB reached epidemic levels mainly in winter and spring. All cases of RSV were identified to be subtype A. PIV infection was mainly caused by PIV-3. The positive rate of HCoV-OC43/HKU1 infection was significantly higher than that of HCoV-229E/NL63. The leading five viruses that resulted in ILI Shijiazhuang in 2011 were HEV, ADV, RSV-A, HRV, and FluA, and these viruses have different epidemiological features.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Influenza, Human ; epidemiology ; virology ; Male ; Middle Aged ; Respiratory Tract Infections ; epidemiology ; virology ; Virus Diseases ; epidemiology ; virology ; Viruses ; classification ; genetics ; isolation & purification ; Young Adult

Single clones of Leptospirillum ferrooxidans were obtained on bilayer solid medium plate by incorporating Het-erotroph Acidiphilium SJH into the underlayer broth medium. The morphologies of the bacteria were investigated using electronic microscope.

AIM:To evaluate the effect of microRNA-155(miRNA-155) on the regulation of angiogenesis in diabetic rats with cerebral ischemic injury .METHODS: Adult male Sprague-Dawley rats were randomly divided into 5 groups:sham group, cerebral ischemia group , diabetic cerebral ischemia group , diabetic cerebral ischemia +miRNA-155 inhibitors group and diabetic cerebral ischemia +scramble group .Diabetes model was made by injection of streptozocin and permanent cerebral ischemic model was developed by suture-occluded method .The scores of neurological deficit and infarct volume were estimated at 24 h after cerebral ischemia .miRNA-155 level was detected by real-time polymerase chain reaction.The expression of platelet endothelial cell adhesion molecule-1 ( PECAM-1/CD31 ) and vascular endothelial growth factor ( VEGF) was detected by Western blotting .RESULTS:miRNA-155 inhibitor significantly reduced miRNA-155 levels in the ischemic cortex (P<0.05), improved the scores of neurological deficit , reduced infarction size and up-regulated the levels of CD31 and VEGF (P<0.05).CONCLUSION:miRNA-155 has a critical role in the regulation of angiogenesis in diabetic rats with cerebral ischemia .Down-regulation of miRNA-155 using miRNA-155 inhibitor attenuates brain infarct injury in diabetic rats .