Readmission rate and related risk factors of ischemic stroke patients after 3 months of discharge in Southwest China

@#Background: Readmission after stroke is common, but limited data is available in West China. This study aimed to assess the frequencies and influencing factors of unplanned readmissions within 3 months after hospital discharge. Methods: This was a retrospective study in a single center. In our study, 596 ischemic stroke patients who were admitted to the Department of Neurology of West China Hospital from November 2011 to August 2012 were enrolled. Demographic data, disease data and follow–up data were collected at first admission and after 3 months of discharge. The readmission rate and risk factors were calculated. Results: Of 596 ischemic stroke patients, the readmission rate was 19.30% (115/596) within three-months, the top three reasons for readmission were needs of rehabilitation (74/115, 64.35%), recurrence of stroke (14/115, 12.17%), complications (11/115, 9.57%). The readmission was associated with older age, whether patients have indwelling catheter and endotracheal tube and pressure sores. (P < 0.05) Conclusions: The rate of readmission within 3 months in ischemic stroke patients was 19.30%. Greater attention should be paid to the elderly patients and patients with endotracheal tube on discharge to reduce readmission. Extended nursing care is also needed to reduce the readmission rate of patients with ischemic stroke.

Effect of the preoperative Geriatric Nutritional Risk Index on the prognosis of patients with esophageal squamous cell carcinoma after radical resection / 中国肿瘤临床

Objective: To evaluate the prognostic value of the preoperative Geriatric Nutritional Risk Index (GNRI) in patients with esophageal squamous cell carcinoma after radical resection. Methods: Clinicopathological and laboratory data of 315 elderly patients with esophageal squamous cell carcinoma who were older than 60 years and underwent radical resection in Tianjin Medical University Cancer Institute and Hospital from January 2008 to December 2012 were retrospectively analyzed. The GNRI formula was as follows:1.489×serum albumin (g/L)+41.7×(current body weight/ideal body weight). According to the GNRI, patients were divided into the normal and abnormal GNRI groups. The χ2 test was used to analyze the relationship between the GNRI and the clinicopathological char-acteristics of patients. The Kaplan-Meier method was used to analyze the survival rate, and survival analysis was conducted using the Log-rank test. Multivariate survival analysis was conducted using the Cox proportional risk regression model. Results: There were 259 patients in the normal GNRI group (GNRI>98) and 56 patients in the abnormal GNRI group (GNRI≤98). The GNRI was closely correlated with age, tumor location, tumor diameter, serum albumin level, body mass index (BMI), and lymph node metastasis (all P<0.05). The 5-year survival rates in the normal and abnormal GNRI groups were 41.2% and 27.0%, respectively, with statistical significance (P=0.002). Univariate analysis showed that age, tumor diameter, serum albumin level, BMI, GNRI, platelet-lymphocyte ratio, tumor invasion depth, and lymph node metastasis were risk factors for the prognosis of patients with esophageal squamous cell carcinoma (all P<0.05). Multivariate analysis showed that the preoperative GNRI (hazard ratio=0.687, 95% confidence interval: 0.487-0.968, P=0.032) was an independent factor affecting the prognosis of patients with esophageal squamous cell carcinoma. Subgroup analysis showed that the survival rates in the normal GNRI group were significantly higher than those in the abnormal GNRI group (P=0.036 and 0.010, respectively), regardless of lymph node metastasis. Conclusions: The preoperative GNRI is associated with malignant biological behav-ior in elderly patients with esophageal squamous cell carcinoma and can be used as a useful indicator for predicting survival after radi-cal resection.

The impact of pain, anxiety and depression on sleep quality in Chinese patients with neuromyelitis optica spectrum disorders

Background & Objective: Sleep quality in neuromyelitis optica spectrum disorders (NMOSD) were investigated in two recent studies. However, factors affecting sleep quality have not been studied in NMOSD. This study aimed to investigate the prevalence of sleep disorders in Chinese outpatient clinics with NMOSD and its clinical correlates. Methods: We administered Chinese validated self-questionnaires on HRQOL (MSQOL-54), sleep (PSQI), pain (SF-MPQ-2), anxiety (HARS) and depression (HDRS) to 42 patients followed up in our outpatient department. We assessed the relationships between sleep quality with pain, anxiety, depression, gender, age, disability, disease duration, NMO-antibody status and explored the determinants of poor sleep quality. Results: Sixty four percent of NMOSD patients were poor sleepers. Significant correlations were found between duration, disability, pain, anxiety, depression and sleep quality. Disability, depression and the domain of affective descriptors of pain were the three main predictors of poor sleep in NMOSD. Conclusion: This study reveals that poor sleep in NMOSD is common and it decreases physical function of quality of life. It is worthwhile considering exploring adjuvant strategies aimed at controlling pain associated affect, and treatment of depression may help to improve sleep quality in NMOSD.

Clinical trials of pemetrexed combined with carboplatin in treating 40 non-small cell lung cancer patients / 中国肿瘤临床

Objective: Pemetrexed (PEM) is a multi-targeted chemotherapeutic agent for antifolate drugs. PEM has become the standard agent for the second-line treatment of advanced non-small cell lung cancer (NSCLC). This study aims to review and analyze the clinical efficacy and adverse reactions of PEM combined with carboplatin with respect to NSCLC treatment. Methods: A total of 40 patients suffering from NSCLC were selected and confirmed by pathology. On the first day of treatment, the conventional 500 mg/m2 dose of pemetrexed disodium was infused intravenously. On the second day, a combined therapy with carboplatin was conducted based on the conventional dose for a 21-day cycle with at least two cycles for each patient. The therapeutic efficacy and adverse reactions were evaluated and were compared with the proposed regimen of gemcitabine (GEM) combined with carboplatin. Results: After two cycles of the treatment, the curative effects of the PEM and GEM groups were 50% and 45%, respectively. The main adverse reactions are bone marrow suppression and gastrointestinal reactions. The incidence rates of bone marrow suppression, gastrointestinal reactions, amisulpride/AST, urea nitrogen, rash, and hair loss were obviously lower in the PEM group than in the GEM group. Statistically significant differences in adverse reaction were found between the two groups (P<0.05). Conclusion: The use of the combination regimen of PEM with carboplatin showed significantly more clinical effects and less adverse reactions for NSCLC treatment.

Effects and mechanism of different doses of rosuvastatin on expression of tissue factor in cultured human monocyte-macrophage cells induced by oxidized low density lipoprotein / 中国医师杂志

Objective To investigate the effects and mechanism of different doses of rosuvastatin on expression of tissue factor(TF) in cultured human monocyte-macrophage cells which were induced by oxidized low density lipoprotein (ox-LDL).Methods The human monocyte-macrophage cells were divided into seven groups:control group,ox-LDL group,poly-insine monophosphate group,different doses of rosuvastatin group(0.01 μmol/L,0.1 μmol/L,1 μmol/L,5 μmol/L).The expression of LOX-1 mRNA and TF mRNA was assayed by RT-PCR.The enzyme-linked immunosorbent assay was performed to determine the protein concentration of TF.Results Effects of different doses of rosuvastatin on expressions of LOX-1mRNA,TF mRNA and TF in cultured human monocyte-macrophage cells induced by ox-LDL:comparison among seven groups,the difference was statistically significant (F =91.334,58.833,103.552,P ＜0.05).Compared with control group,the expressions of LOX-1 mRNA,TF mRNA and TF were increased in the ox-LDL group[(3.25156 ± 0.15772) vs (1 ±0) ;(2.522451 ±0.138967) vs (1 ±0) ;(207.7233± 1.154701)ng/L vs (184.8467 ± 0.871799)ng/L],and they were in a concentration-dependent manner (P ＜ 0.05).Compared with the PolyⅠ group and the different doses of rosuvastatin group,the expressions of LOX-1 mRNA,TF mRNA and TF were in the ox-LDL group,and the different doses of rosuvastatin were decreased by dose-dependent manner.It was in a concentration dependent manner (P ＜ 0.05).Different doses of rosuvastatin were compared between groups (between each group P ＜ 0.05),the difference between each two groups was statistically significant (P ＜ 0.05).Conclusions LOX-1 may be responsible for the expression of TF in Human monocyte-macrophage cells induced by ox-LDL.Rosuvastatin by dose dependent manner and by means of ox-LDL reduced monocyte-macrophage LOX-1 mRNA and TF mRNA expressions,which reduced expression of TF.

Molecular epidemiology of group A rotavirus diarrhea in children in Shanghai / 中华传染病杂志

ObjectiveTo explore the molecular epidemiological characteristics of group A rotavirus diarrhea in children in Shanghai and to provide the background data for the implementation of rotavirus vaccination.MethodsA total of 910 stool samples were collected from the outpatient children with acute diarrhea from August 2008 to July 2009.Group A rotavirus was detected by usingcommercial colloidal gold device.Rotavirus strains were characterized for G and P genotypes using the nested reverse transcription polymerase chain reaction (RT-PCR).ResultsGroup A rotavirus was detected in 268(29.4％) out of 910 stool samples.Rotavirus infection was found year-round and the peak season was from October 2008 to January 2009,with the detection rates ranging from 38.3 ％ to 70.5％.Ninety-one percent of children (244 cases) with rotavirus-associated diarrhea occurred in children ＜3 years of age.The detection rate of rotavirus was highest (36.6％) in children aged 12-23 months.Among the 268 group A rotavirus-positive strains,G1 was the most common G genotype (65 strains),accounting for 24.3％,followed by G3 (40 strains,14.9％),G mixed genotypes (37strains,13.8 ％),G2 (27 strains,10.1％),G9 (14 strains,5.2％),G4(5 strains,1.9％),other G types (5 strains,1.9％),and unclassified G type (75 strains,28.0％).P[8] and P[4] were the most common P genotypes,accounting for 54.9％ (147 strains) and 11.9％ (32 strains),respectively,followed by P mixed genotypes (6 strains,2.2％) and other P genotypes (4 strains,1.5％),unclassified P type (79 strains,29.5％).The G/P genotype combinations were found as follows:G1P [8] (13.4％),G3P[8] (13.4％),GmixP[8] (10.1％),G1P[4] (8.2％),G9P[8] (2.2％),G2P [4] (1.9％),G1Pmix (1.9％).ConclusionsGroup A rotavirus is a major causative agent of diarrhea in infants and young children in Shanghai.The peak season of rotavirus infection appears in fall and winter.The currently licensed rotavirus vaccines cover the majority of genotypes of rotavirus strains prevailing in Shanghai.

Comparison of drug resistance mutations associated with hepatitis B virus covalently closed circular DNA and relaxed circular DNA / 中华传染病杂志

Objective To detect nucleos(t)ide-resistance mutations in hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) isolated from hepatocytes of patients with chronic HBV infection and to analyze the correlation between the mutations found in cccDNA and relaxed circular DNA (rcDNA). MethodsForty patients with chronic HBV infection were investigated. Preoperation serum samples and non-tumor liver tissues were collected.Intrahepatic HBV cccDNA and rcDNA were selectively extracted by co-precipitation of sodium dodecyl sulphate-protein and QIAamp DNA Mini Kit, and further purification with plasmid-safe ATP-dependent DNase (PSAD).Thereafter,cccDNA were amplified by selective polymerase chain reaction (PCR) or nested PCR using the primers spanning both the two gaps in HBV genome and covering the common mutations associated with nucleoside analogues resistance (rt169- rt250).Intrahepatic HBV rcDNA and pre-operation serum HBV rcDNA were also extracted and then amplified by PCR.The PCR products were then purified and sequenced.Results Among the 40 patients,intrahepatic HBV cccDNA were detected in 31 patients,and HBV rcDNA were detected in liver samples of 35 patients and pre-operation serum samples of 21 patients. The PCR products amplified from these samples were all successfully sequenced.rtM204Ⅰ mutation was detected in intracellular HBV cccDNA,rcDNA and serum HBV rcDNA in 2 patients.Both rtM204Ⅰ and rtQ215H were detected in intrahepatic HBV cccDNA and rcDNA in 2 patients,while no corresponding mutation was observed in serum HBV rcDNA of these two patients.Three variants including rtM204V,rtM204V and rtV173L-rtL180M-rtM204V were detected in serum HBV rcDNA in 3 patients,while corresponding mutants were not detected in intracellular HBV cccDNA or rcDNA of these patients.Condsions The results suggest that antiviral nucleos(t) ide resistance mutations can be found in HBV cccDNA in chronic hepatitis B patients. The dominant resistant mutation found in intrahepatic HBV cccDNA/rcDNA may be different from that in serum HBV rcDNA.

Identification of hepatitis B virus X-interactive proteins in human hepatocellular carcinoma cell lines HepG2 by using proteomics technique / 中华传染病杂志

Objective To identify hepatitis B virus X-interactive proteins by comparative proteomics method and to understand the molecular mechanism of HBx in the development of hepatocellular carcinoma(HCC).Methods Two-dimensional gel electrophoresis(2-DE)was used to separate the total proteins of HBx-transfected human hepatoma cell lines HepG2-Px and its parental cell lines HepG2-P_0.PDQuest software was applied to analyze 2 DE images.The differentially expressed protein spots between the two cell lines were identified by matrix-assisted laser desorptionionization time of flight mass spectrometry(MALDI-TOF-MS)and electrospray ionization tandem mass spectrometry(ESI-Q-TOF).Then,the differential expression levels of some identified proteins were determined by Western blot.The data were compared using t test.Results The well-resolved,reproducible 2-DE patterns of HepG2-Px and HepG2-P_0 total proteins were established.A total of 32 differential proteins were identified in HepG2-Px cell,including 25 up-regulated proteins,such as heat shock protein(HSP)90AB1,Bcl-2 associated athanogene(BAG)-2,nucleophosmin(B23),chloride intracellular channel(CLIC)-1,matrix metalloproteinase(MMP)-3,melanoma antigen(MAGE)-12,and 7 down-regulated proteins,such as Wnt-5a.The differential expression levels of some proteins between the two cell lines were confirmed by Western blot analysis.Conclusions Most of the identified proteins are involved in many processes,such as transcription,signal transduction,cell proliferation,cell cycle regulator,apoptosis,DNA repair,metabolisms and immunity.These differential proteins may play a role in tumor genesis and HC development.The data are valuable for further study on the role of HBx in human hepatocellular carcinoma.

Effects of hepatitis B virus X protein on the induction of cytochrome P450 3A4 in HepG2 cells / 中华传染病杂志

Objective To investigate the effect of hepatitis B virus X protein (HBx) on the induction of eytochrome P450 (CYP) 3A4 by 1α, 25-(OH)2D3 in HepG2 cells in vitro. Methods HepG2 cells were transiently transfected with plasmid pEGFP-N1 (control) or co-transfected with recombinant HBx eukaryotic expression plasmid pcDNA3-X and pEGFP-N1. All HepG2 cells were divided into four groups: control group (without transfection), plasmid pEGFP-N1 transfection group, plasmid pEGFP-N1 transfection plus 1α ,25-(OH)2D3 group, plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α ,25-(OH)2D3 group. The expression of CYP3A4 in HepG2 cell was induced by 0.35 μ mol/L 1α ,25-(OH)2D3 for 72 h, and mRNA levels and protein levels of CYP3A4 in the cells were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and Western-blot assay, respectively. The comparison between groups was done by F test. Results CYP3A4 mRNA level in plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α ,25-(OH)2D3 group was 1.52 folds of control group, while that in plasmid pEGFP-N1 transfection plus 1α, 25-(OH)2D3 group was 3.97 folds (F= 4.72, P<0. 05). Similarly, intracellular CYP3A4 protein expression in plasmid pcDNA3-X and pEGFP-N1 co-transfection plus 1α , 25-(OH)2D3 group increased to 2.1 folds of control group, while that in plasmid pEGFP-N1 transfection plus 1α,25-(OH)2D3 group increased to 5.9 folds (F=4.68, P<0.05). Conclusion HBx interferes with the induction of CYP3A4 by 1α , 25-(OH)2D3 in HepG2 cell line, which suggests that HBx has suppressive effect on the expression of CYP3A4.

Detection of hepatitis B virus covalently closed circular DNA in serum of patients with hepatitis B / 中华传染病杂志

Objective To determine whether hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) could be detected in serum of patients with hepatitis B and evaluate the related factors and clinical significances. Methods Fifty-seven patients, including 26 with mild chronic hepatitis B (CHB) and 31 with severe hepatitis B (SHB) were enrolled. Prothrombin time (PT), hepatic biochemical indexes, serum markers of hepatitis virus, serum total HBV DNA and HBV cccDNA of every patient were detected after hospitalization. Factors associated with the detection rate of serum HBV cccDNA were analyzed using Logistic stepwise regression. Results Serum HBV cccDNA was detected in 13 patients with SHB and 1 with mild CHB, and serum levels of HBV cccDNA were varying from 1.25 × 103 to 4. 88 × 104 copy/mL. The detection rates were significantly different between the two groups (P=0. 0014). The sensitivity and specificity of SHB diagnose by serum HBV cccDNA detection were 41.94% and 96.15 %, respectively. Logistic regression analysis showed that the detection rate of serum HBV cccDNA was associated with PT (X2 = 7. 2192, P= 0. 0072), while not associated with age, sex, total serum HBV DNA, total bilirubin or alanine aminotransferase (ALT). Conclusion Serum HBV cccDNA could be detected in some of the patients with SHB, whic hmay be considered as one of the diagnostic indexes for SHB,

Follow-up study on the etiology of acute hepatitis B in adults / 中华传染病杂志

Objective To understand the etiology of acute hepatitis B (AHB) in adults and investigate the mechanisms of hepatic injury and viral clearance in AHB. Methods One hundred and twenty adult AHB patients were enrolled. Epidemiological and clinical data were collected from the case history records or face-to-face inquiry, and serum samples were collected during hospitalization and follow-up. To observe dynamic patterns of AHB etiology, the markers of hepatitis B virus (HBV) were detected by enzyme-linked immunosorbent assay (ELISA); the level of HBV DNA and HBV genotype were determined by real-time polymerase chain reaction (PCR). Enumeration data were analyzed by non-parametric rank sum test. Comparison between groups was done by t test and that between rates of samples was done by Pearson χ2 test. Results Serum HBV DNA was positive in 48.33% of patients at the time of diagnosis with mean level of 9.84×04 copy/mL, and became undetectable after 12.5 days on average. The median levels of serum alanine aminotransferase (ALT) were 1600 U/L and 1490 U/L in HBV DNA positive and negative groups, respectively (z=-0. 678, P=0. 498). However, the mean levels of serum ALT were (2058±123) U/L and (1393±139) U/L in groups of HBV DNA<1×104 copy/mL and>1×104 copy/mL, respectively, which was significantly different (t=-2.17, P=0. 049). Genotype B accounted for 52.5%, genotype C 42.5 and genotype B and C mixed type 5.0% in 58 patients with HBV DNA positive. Eight patterns of serum HBV markers were presented at first visiting. HBsAg(+), HBeAg(+), anti-HBc(+), anti-HBc IgM(+) and HBsAg(+), anti-HBe(+), anti-HBc(+), anti-HBc IgM(+) were the most common patterns, which accounted for 38.3% and 30.0%, respectively. The dynamic patterns of serum HBV markers of 28 AFIB patients were prospectively followed up. The rate of serum FIBsAg loss was 100. 0% and the median time of negative-conversion was 3 weeks. The cumulative positive rate of anti-HBs was 85.7% after 52 weeks of follow-up. The rate of serum HBeAg loss was 100.0%. HBeAg was negative in 53.6% of patients at first visiting and the rest of patients achieved negative within 4 weeks after onset. The positive rate of anti-HBe was 82.1% during 52 weeks of follow-up. Total anti-HBc (including IgG and IgM) was keeping positive in all patients within 52 weeks, and the negative rate of anti-HBc IgM was 39. 3% after followed up for 52 weeks. Conclusions Rapid HBV clearance andserum HBV marker conversion are significantly different between AHB and chronic hepatitis B.

Effects of chronic hepatitis B virus infection on human hepatic cytochrome P450 2C9 / 中华传染病杂志

Objective To investigate the effects of chronic hepatitis B virus (HBV) infection on human hepatic cytochrome P450 2C9 (CYP2C9).Methods Liver tissue samples and blood samples were obtained from 10 patients with chronic HBV infeetion and 10 healthy controls.CYP2C9 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.The activity of CYP2C9 was detected utilizing high performance liquid chromatography (HPLC).The expressions of CYP2C9 mRNA and protein were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blotting.The data were analyzed by t test.Results All the liver samples showed CYP2C9 wild-type (*1*1),while CYP2C9 (*2) and CYP2C9 (*3) were not detected.The maximum velocity (Vmax) of CYP2C9 in patients chronic HBV infection and healthy controls were (263.5±66.4) μmol/L and(284.6±85.9) μmol/L,respectively (t=0.614,P=0.5471).The expression of CYP2C9 mRNA in patients with chronic HBV infection (0.39±0.28) was significantly lower than that of healthy controls (0.65±0.13) (t=2.628,P=0.0171).Accordingly,the protein expression in patients with chronic HBV infection (0.26±0.13) was lower than that of healthy controls (0.60±0.19) (t=4.688,P=0.000 2).Conclusion The expressions of CYP2C9 mRNA and protein are decreased in chronic HBV infection which may down-regulate the enzyme activity.

Inhibition effect of total glucosides of Picrorhiza on replication of hepatitis B virus in vitro / 中华传染病杂志

Ohjective To observe the inhibition effect of total glucosides of Picrorhiza on hepatitis B virus covalently closed circular DNA (HBV cccDNA) in HepG 2.2.15 cell line. Methods HepG 2.2.15 cells were incubated with culture medium containing 50 mg/L of picrosides or 5 mg/L of adefovir dipivoxil for 2 or 5 days. HBV DNA in the supernatant, intracellular cccDNA, relaxed circular DNA (rcDNA) and pregenomic RNA (pgRNA) were quantified by specific real-time polymerase chain reaction (RT-PCR) and inhibition rates were calculated. The means were compared by t test. Results After treated with picrosides for 2 and 5 days, the inhibition rates of HBV DNA in thesupernatant were 49. 74% (t=4.723, P<0.05) and 79.48% (t = 7.512, P<0.05), respectively. The inhibition rates of intracellular cccDNA were 43.55% (t = 5.216, P<0.05) and 56.43% (t=7.262, P<0.05), respectively, while those of intracellular rcDNA were 43.39% (t=4.137, P<0.05) and 63.86% (t=7.861, P<0.05), respectively, and those of intracellular pgRNA were 54.72% (t=4.532, P<0.05) and 56.08% (t=4.833, P<0.05), respectively. Comparatively, after treatment with adefovir dipivoxil for 2 and 5 days, the inhibition rates of HBV DNA in the supernatant were 25.56% (t=2.874, P<0.05) and 92.44% (t =10.276, P<0.05), respectively. Those of cccDNA were 18.54% (t=2.736, P<0.05) and 47.19% (t=6.852, P<0.05), respectively. Those of rcDNA were 21. 20% (t=3.206, P<0.05) and 71.47% (t=8.332, P<0.05), respectively, pgRNA were 11.14% (t=1.761, P>0.05) and 37.61%(t=3.632, P<0.05) respectively in HepG2.2.15 cells. Conclusions Pierosides may inhibit the replication cycle of HBV, including the formation of cccDNA in HepG 2.2.15 cells. The mechanism of pierosides on cccDNA may differ from adefovir dipivoxil's due to its earlier inhibition time phase.

Effect of oxymatrine on the activity of HBV polymerase in vitro / 中华临床感染病杂志

Objective To determine the effect of oxymatrine on the activity of HBV DNA polymerase in vitro. Methods Hepatitis B virus particles were purified from supernatant of cultured HepG2.2.15 cells by uhracentrifugation, and then were mixed with reaction buffer containing NP-40, β-mercaptoethanol, 32P-labelled nucleoside triphosphate (dCTP), MgCl2, and different concentrations of oxymatrine ( 1000 μg/ml, 800 μg/ml, 600 μg/ml, 400 μg/ml and 200 μg/ml) or adefovir dipivoxil ( 100 μg/ml, 80 μg/ml and 60 μg/ml, 40 μg/ml and 20 μg/ml). After incubation at 37 ℃ overnight, proteinase K was added to the reaction system for digestion and 35 μl of samples were spotted onto DE81 paper. Activities of endogenous polymerase in HBV particles were assessed by determining the radioactivity of 32P-labelled dCTP incorporated in the plus-strain of viral DNA. Results Compared with the blank control, the activity of endogenous polymerase in HBV particles treated with different doses of oxymatrine varied from 103% to 107%, and it varied from 91% to 101% when treated with different doses of adefovir dipivoxil. No significant difference was observed among treated groups and the control (P > 0. 05 ). Conclusion No direct inhibitory effect of oxymatrine on the activity of HBV polymerase was observed in vitro.

Several relationships needed to be well handled during the teaching of"Medical microbiolo-gy"and"Medical immunology"courses for undergraduate students / 中华医学教育探索杂志

Both of "Medical Microbiology"and "Medical Immunology"are two important basic and required medical courses for college students.There is something worth discussing in some respects,including the composing of teaching materials,designing of curriculum,key points of teaching and practical application.It's necessary to deal with the relationships well between the teaching effects and the issues such as arrangements of class period,selection of teaching material and the design of teaching.Particularly,we should emphasize close connection between the basic and clinical course.

Developing and Emphasizing Harmonious Relationship between Teaching and Learning in Clinical Medical School / 中华医学教育探索杂志

In clinical medical school,disharmony between teaching and learning has been existing which affects the teaching quantity as well as the cultivation of talents.Personal suggestions about the ways to deal with these problems are given in this paper.

3′-terminus shifted bases degeneracy primers increasing sensitivity of polymerase chain reaction / 第二军医大学学报

To reduce the risk of 3′-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU,WD) was designed to amplify a fragment of HBV DNA P gene by PCR. Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3′-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2, MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3′-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70.4%(19/27) and 85.2%(23/27) respectively (P<0.05). (2) The sequencing analysis of the PCR products suggested that the 3′-terminal mismatch of primers caused false negative in the PCR detection. Conclusion: When amplifying the variable region of DNA, the false negative result can be avoided by using 3′-terminus shifted degeneracy primers.

Detection and genetic analysis of TT virus in hemodialysis patients / 第二军医大学学报

Objective: To study the prevalence and pathogenesis of TT virus (TTV) in hemodialysis patients. Methods: Serum TTV DNA was tested in 69 hemodialysis patients from our hospital by nested-PCR using primers from a conservative region of TTV genenome, genetic analysis and detection of hepatitis C virus antibody (anti-HCV) and the levels of alanine aminotransferase (ALT) were also carried out simultaneously. Results: The overall prevalence of TTV viremia was 27.5%. The PCR-amplified gene fragment from one patient was sequenced, and its gene sequence homologies with GH1,TA278, TTVCHN1 and TTVCHN2 ranged from 89% to 100%, its deduced amino acid sequence ranged from 87% to 100%. There was no significant difference of TTV prevalence between anti-HCV positive and negative patients. No significant elevation of ALT was found in all patients. Conclusion: High prevalence of TTV infection is found among hemodialysis patients, and TTV infection has no significant association with HCV infection or elevation of ALT.

FEVER OF UNKNOWN ORIGIN: A RETROSPECTIVE ANALYSIS OF 107 CLINICAL CASES / 解放军医学杂志

To investigate causes and clinical picture of fever of unknown origin (FUO), and to sum up the experiences in diagnosis of FUO, the medical records of 107 patients with FUO were reviewed retrospectively. Specific causes were identified in 99. 1% of these patients, including infections in 50 patients(46. 7%) , rheumatic problems in 22(20. 6%) and malignancies in 17(15. 9%). The main pathogens responsible for the infections were pyogenic bacteria(72. 0% , 36/50) and M tuberculosis (18. 0% , 9/50), mostly extrapulmonary. Lymphatic and he-mopoietic tissue neoplasms were the main forms of malignancy (88. 2%, 15/17), including histiocytosis, malignant lymphoma and leukemia. Drug fever was another common cause of FUO, accounting for 8. 4% in our series.

Quantitative detection of hepatitis B virus cccDNA in infected hepatocytes / 中华传染病杂志

Objective To establish a method for quantitative detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA ) in infected cells. Methods The transfected cell line HepG2.2.15 which can consistently produce Dane particles was maintained in DMEM containing 380 ?g/ml G418 and 10% fetal bovine serum. Cells in the exponential period were harvested from flasks, then intracellular HBV cccDNA was extracted from pellet containing 1?10~6 cells with mini plasmid extraction kit (QIAGEN).The extraction product was further purified by mung bean nuclease to remove HBV relaxed circular DNA possibly remained. HBV cccDNA was quantitatively detected by fluorescent PCR with selective primer set and Taqman MGB probe. Culture medium before exponential period, HBV DNA positive and negative serum samples from patients with chronic hepatitis B (mild) were amplified simultaneously to test the specificity of the fluorescent PCR method. Plasmids containing whole HBV genome were amplified with the same primer set and fluorescent probe to determine the sensitivity of the method. Results HBV cccDNA was detectable in HepG2.2.15, and the average quantity was 18 copies per cell approximately. No detectable fluorescent signal was observed when culture and serum samples were amplified. The detectable HBV cccDNA was as low as 10~3 copies per ml at least by this method. Conclusions This method is convenient, highly specific and highly sensitive. It can be utilized in the quantitative detection of intracellular HBV cccDNA as well as in the screening and evaluation of antiviral agents.