Objective To explore the efficacy of sequential or combined amphotericin B (AmB) and fluconazole (FCZ) therapy on a 5 flucytosine based regimen in non-acquired immunodeficiency syndrome (AIDS)-related cryptococcal meningitis.Methods A tatal of 117 cases of non-AIDS-related cryptococcal meningitis treated with 5-flurocytosine-based regimens were retrospectively divided into five groups:AmBgroup (n=38),FCZ group (n=25),FCZ and AmB sequential group (n=18),AmB and FCZ sequential group (n=15),AmB and FCZ combination group (n=21).The number in cerebrospinal fluid (CSF) of the five groups were compared.Statistical analyses included t test,oneway analysis of variance,K-independent samples test and chi-square test.Results Intracranial pressure of AmB group,FCZ group,FCZ and AmB sequential group,AmB and FCZ sequential group,AmBandFCZ combination group were (208.6±75.1),(191.5±94.5),(185.0±76.3),(201.9±69.7) and (223.1±89.3) mm H2O (1 mm H2O=0.0098 kPa),respectively,and the differences were not statistically significant (F=0.611,P =0.656).Median cryptococcus counts in CSF of the five groups were 0,10,0,3 and 0/mL,respectively,with no statistical significance (x2 =7.638,P-0.090).CSF protein levels of the five groups were 0.55,0.69,0.67,0.53 and 0.96 g/L,respectively,with no significant differences among groups (F=7.063,P=0.133).The cure rates of the five groups were 55.3％ (21/38),32.0％ (8/25),9/18,6/15 and 47.6％ (10/21),respectively;progression rates or mortality of the five groups were 28.9％ (11/38),44.4％ (11/25),5/18,4/15and 23.8％ (5/21),respectively; and the differences among cure rates (x2 =3.638,P=0.457) and progression rates or mortality (x2-2.785,P =0.604) were not statistically significant.Conclusion FCZ or AmB alone,sequential or combined therapy were all effective in the treatment of cryptococcal meningitis.

ObjectiveTo investigate the therapeutic effects of patients with non-acquired immunodeficiency syndrome ( AIDS )-related cryptococcal meningitis treated with fluconazole and flucytosine.MethodsA retrospective study was conducted including 52 cases of non-AIDS-related cryptococcal meningitis,which were divided into two groups:the amphotericin B combined with flucytosine group (n =32) and the fluconazole combined with flucytosine group (n =20 ). The comparison between groups was done by t test and chi-square test.ResultsThe intracranial pressure of the amphotericin B combined with flucytosine group and the fluconazole combined with flucytosine group were (221.9±76.2) and (213.4±99.2) mm H2O,respectively (t=0.302,P＞0.05) ; the cryptococcus counts in the cerebrospinal fluid (CSF) were (351 ± 1180) and (508 ± 943)/mL,respectively (t=- 0.473,P＞0.05) ; the CSF protein levels were (0.754 ± 0.726) and (0.649 ±0.308) g/L,respectively (t=0.578,P＞0.05) ; the CSF white blood cell (WBC) counts were (16±25) × 106/L and (28±32) × 106/L,respectively (t=-1.348,P＞0.05).The cured rates were 59.38％ (19/32) and 35.00％ (7/20),respectively in the amphotericin B combined with flucytosine group and the fluconazole combined with flucytosine group,and the improved rates were 6.25％(2/32)and 25.00％ (5/20),respectively,the uncured or mortality rates were 34.38％ (11/32) and 40.00％ (8/20),respectively,and the difference were not significant of cured rates (x2 =2.925,P＞0.05) and uncured or mortality rates (x2 =0.168,P＞0.05) between two groups.ConclusionFluconazole combined with flucytosine also has a good effect on the treatment of cryptococcal meningitis.

Objective To evaluate the efficacy of entecavir treatment on hepatitis B patients with acute-on-chronic liver failure. Methods Eighty-four hepatitis B patients with acute-on-chronic liver failure were treated with entecavir 0.5 mg daily and Other routine drugs. Another 99 hepatitis B patients with acute-on-chronic liver failure were treated with only routine drugs as control. The survival, liver functions, hepatitis B virus (HBV) DNA level, prothrombin time (PT) were observed. The survival rates of patients with early, middle or late stage of liver failure were analyzed. The comparison of rates were done using chi-square test. The numeration data were compared by t test. The survival rates were compared using Kaplan-Meier method. Results Among patients with early stage of acute-on-chronic liver failure, the survival rate in treatment group was 63.3% (31/49), which was significantly higher than that in control group (39.7%, 23/58) (χ2=5.923, P=0.015). Among patients with middle stage of acute-on-chronic liver failure, the surviral rate in treatment group was 63.0% (17/27), which was significantly higher than that in control group (35.1%, 13/37) (χ2=4.854, P=0.028). Among patients with late stage of acute-on-chronic liver failure, four out of eight cases survived in treatment group, while one out of four cases survived in control group. In patients with serum total hilirubin (TBil) level > 342 μmol/L, the survival rate was 56.0% in treatment group, which was significantly higher than that in control group (26.8%) (χ2=9.351,P=0.002). At week 4 of the treatment, the HBV DNA reduction in treatment group was 3. 95 lg copy/mL, which was higher than that in control group (1.78 lg copy/mL) (t=5.847, P=0.001). Conclusions Entecavir treatment could improve the survival rate of hepatitis B patients with early or middle stage of acute-on-chronic liver failure. And the further study with larger population is needed in patients with late stage of liver failure. In addition, entecavir therapy could also improve the survival rate of patients with TBil >342 μmol/L.

Objective To compare the advantages and disadvantages of retroperitoneal laparoscopic sur -gery and open surgery in treatment of adrenal adenoma , in order to provide the clinical evidence for the selection of surgical treatment of adrenal adenoma .Methods 120 cases undergoing surgical treatment from Jan .2005 to Dec.2010 were randomly divided into laparoscopic group and open surgery group .Operation indicators , postop-erative recovery indicators , postoperative clinical manifestations , success of operation , complications and follow-up data were evaluated and compared between the 2 groups.Results 3 cases in laparoscopic group and 4 cases in open surgery group withdrew.The operation time, intraoperative blood loss, and blood transfusion cases were (89.6 ±18.4)min vs (124.2 ±27.3)min, (70.2 ±17.2)ml vs (205.7 ±51.9)ml, and 8 cases vs 20 cases in laparo-scopic group and open surgery group .Extubation time, starting feeding time, time of getting out of bed, and length of hospital stay were (2.8 ±1.3)d vs (4.2 ±2.0)d, (1.7 ±1.1)d vs (2.9 ±1.4)d, (2.6 ±1.2)d vs (5.1 ± 3.1)d and (6.2 ±3.1)d vs (13.5 ±5.4)d in laparoscopic group and open surgery group .Postoperative analge-sic using cases and drainage volume were 11 cases vs 43 cases, and(15.4 ±4.2)ml vs(45.1 ±12.7)ml in lapa-roscopic group and open surgery group .The difference had statistical significance (P＜0.01).The cost was simi-lar between the 2 groups(P>0.05).The open surgery group had no death or relapse within 1 year, while 5 cases in laparoscopic group converted to laparotomy , with 1 case of death and 2 cases of relapse within 1 year.The complications incidence was 3.51%(2/57)in laparoscopic group, obviously lower than 16.07%(9/56)in open surgery group(P＜0.01).Conclusions Compared to the traditional open surgery, retroperitoneal laparoscopic sur-gery has advantages of less trauma, fewer complications, and faster postoperative recovery in treatment of adrenal ade-noma.Experience and skills of surgeon need to be improved to reduce the possibility of failure or recurrence .

Objective To understand the clinical features of hepatitis B virus(HBV)/hepatitis C virus(HCV)coinfected patients with different virological profiles.Methods The clinical data of 186 patients with HBV/HCV coinfection from May 1999 to May 2010 in the Third Affiliated Hospital of Sun Yat-Sen University were analyzed retrospectively.The demographic data,epidemiological data,laboratory results and pathological index were analyzed.The statistical analysis was done using t test and chi square test.Results A total of 186 patients were divided into 4 groups:66(35.5%)in HBV DNA(-)/HCV RNA(-)group,8(4.3%)in HBV DNA(+)/HCV RNA(+)group,68(36.6%)in HBV DNA(+)/HCV RNA(-)group and 44(23.7%)in HBV DNA(-)/HCV RNA(+) group.The gender composition,complication incidence,transmission among drug users,alanine aminotransferase(ALT)level,total bilirubin(TBil)level,prothrombin activity(PTA)and hapatitis B e antigen(HBeAg)negative rate were all significantly different among four groups(F or x2=11.578,8.451,11.738,2.669,5.102,4.254 and 18.413,respectively;all P<0.05).In groups of HCV RNA(-)and HCV RNA(+),the proportions of patients infected through drug abuse were 49.3%and 23.1%,respectively(x2=9.987,P:0.002)and blood transfusion transmission were 29.9%and 46.2%,respectively(x2=4.412,P=0.036).When HBV DNA was negative,the median ALT levels in HCV RNA(-)and HCV RNA(+)patients were 177 U/L and 62 U/L,respectively(t=2.200,P<0.05),median TBil levels were 133 μmol/L and 20μmol/L,respectively (t=3.608,P<0.05)and PTA were 70.6%±27.7%and 83.3%±27.8%,respectively(t=-1.982,P<0.05).The HBeAg negative rate was not affected by HCV RNA levels(t=0.707,P>0.05).The HBeAg negative rate in HBV DNA(-)patients was 85.5%,which was higher than that in HBV DNA(+)patients(59.2%)(x2=16.393,P<0.05).Conclusions HBV DNA(+/-)/HCV RNA(-)profile were major components in HBV/HCV confection.HBV DNA level is related to disease progression and prognosis,but not relate to disease severity.Liver function damage and disease severity are aggravated with HCV RNA level decreases.HBV DNA level is related to HBeAg negative rate,while HCV RNA level is not related to HBeAg seroconversion rate.

Objective To investigate the level of serum macrophage migration inhibitory factor (MIF) and its correlation with serum precollagen Ⅲ peptide (PⅢP) and tissue inhibitor of metalloproteinase (TIMP)-1 in patients with chronic hepatitis B (CHB) and hepatitis B virus (HBV)-related cirrhosis. Methods Forty-four CHB patients (hepatitis B group), 44 patients with HBV-related cirrhosis (cirrhosis group) and 30 healthy controls (control group) were enrolled in this study. The venous blood was collected and MIF level was detected by enzyme-linked immunosorbent assay (ELISA). Correlations between MIF and PⅢP, TIMP-1 were analyzed in observed groups. Comparison between groups was done using t test. The correlations between MIF level and alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), plasma thromboplastin antecedent (PTA), PⅢP and TIMP-1 were analyzed by rectilinear correlation. Results The levels of serum MIF, PⅢP and TIMP-1 in CHB group and cirrhosis group were all significantly higher than those in control group (t=12.87,5.28, 10.98,t=11.22,14.84,11.17;all P<0.05), while there were no significant differences between CHB group and cirrhosis group (t= -1.05,1.52,--2.07;all P>0.05). There was no correlation between MIF level and ALT, AST, TBil and PTA. MIF level in CHB patients with hepatitis B e antigen (HBeAg) positive and high viral load were both higher than that in patients with HBeAg negative and low viral load. MIF level was both positively correlated with PⅢP level in CHB group and cirrhosis group (r=0. 603, P<0.05 and r=0. 415, P<0. 05, respectively). MIF level was also positively correlated with TIMP-1 level in CHB group (r=0. 458, P<0.05), while not correlated in cirrhosis group (r=0. 210, P>0.05). Levels of PⅢP and T1MP-1 were both correlated in CHB group and cirrhosis group (r=0. 849, P< 0.05 and r=0. 424, P<0.05, respectively). Conclusions The levels of serum MIF are significantly increased both in patients with CHB and cirrhosis. The early production of MIF might be related with viral replication, but not with liver function. MIF participates in formations of hepatitis, liver fibrosis and cirrhosis, which could reflect the degree of liver cirrhosis.

OBJECTIVETo evaluate the feasibility of oral cytokine gene therapy against tumor using live attenuated Salmonella as a vector.

METHODSA live attenuated AraA- autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1 and pCMVmIL-12 administered orally in BALB/c and C57BL/6 mice. After 6 weeks, the mice were challenged with 4T1 or Lewis tumor cells, respectively, and flow cytometry and confocal microscopy were used to detect the expression of green fluorescence protein (GFP) in the tissues. PCR and ELISA were performed to detect the integration and expression of mIL-12 gene, and the survival time of the mice was also recorded.

RESULTSGFP expression and mIL-12 gene integration could be detected in the liver, spleen, intestinal, kidney and tumor tissues of the mice. The serum level of mIFN-gamma, mIL-12 increased significantly in mice with oral mIL-12 administration (P<0.05), which resulted in the survival prolongation of the mice as compared with the control mice (P<0.05).

CONCLUSIONOral gene therapy using live attenuated Salmonella can be potentially a simple, effective and above all, safe means for tumor treatment.

Administration, Oral ; Animals ; Carcinoma, Lewis Lung ; therapy ; Flow Cytometry ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Interleukin-12 ; genetics ; metabolism ; Mammary Neoplasms, Animal ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Microscopy, Confocal ; Neoplasms, Experimental ; therapy ; Salmonella typhimurium ; genetics ; immunology ; metabolism ; Vaccines, Attenuated ; immunology

Objective To investigate the therapeutic effects of recombinant yeast hepatitis B virus(HBV)vaccine combined with interferon(IFN)α-1b and determine the rational dosage of HBV vaccine for the further clinical study with larger sample.Methods Two hundreds and sixteen patients with chronic hepatitis B(CHB)were enrolled in this randomized,multi-center,double-blinded and placebo-controlled clinical trial.All the subjects were not treated with antiviral drugs within 6 months and randomly divided into 90μg,60μg and placebo groups with a ratio of 1:1:1.All the patients were intramuscularly administrated with 90μg or 60μg recombinant HBV vaccine or placebo at week 0,2,6,10,14,18,22,respectively.Meanwhile,they were also treated with IFNα-1b 50μg,3 times a wcek for 24 weeks.All patients were followed up for 24 weeks after withdrawal of anti-HBV therapy.Serum HBV DNA level,HBeAg titer and liver function were monitored frequently.Interferon-γ secreting lymphoeytes were detected by Enzyme-linked immunospot(ELISPOT)in part of the patients.Results The serum HBV DNA levels were(6.03±1.79),(5.52±1.82)and(6.29±1.70)log10 copy/mL at week 24 of treatment in high dose,low dose and placebo groups,respectively (P=0.458).And the serum HBV DNA levels were(5.92±1.98),(5.49±1.99)and(6.16±1.76)log10 copy/mL at weck 24 after withdrawal of treatment,respectively(P=0.720).The rates of patients whose HBV DNA＜1×105 copy/mL in these three groups were 30.4%,39.4% and 20.8% at week 24 of treatment,respectively and there was significant difference between high dose group and low dose group(P=0.015).The rate of patients whose HBV DNA＜1×105 copy/mL at week 24 after withdrawal was highest in low dose group,but no significant differences before and after treatment in these three groups(P=0.257).The HBV DNA negative rates were 17.4%,25.4% and 6.9% in these three groups,respectively,which were significantly different(P=0.012).At week 24 of treatment and week 24 after withdrawal of treatment,the alanine aminotransferase normalization rate,HBeAg seroconversion rate were highest in low dose group,but no significant differences in these three groups.ELISPOT positive rates at week 24 of treatment and week 24 after withdrawal of treatment in high close and low dose groups were higher than that in placebo group(P＜0.05).The incidence of adverse events was similar and there was no drug related severe adverse events in each group.Conclusion Recombinant HBV vaccine maybe contribute to anti-HBV therapy and 60μg of dosage seems to be rational.

OBJECTIVETo determine the immune repertoires of peripheral CD4+T cell receptor (TCR) Vb CDR3 in primary biliary cirrhosis (PBC) and analyze TCR diversity and preferred usage at sequence-level resolution.

METHODSARM-PCR and high-throughput sequencing were used to obtain millions of TCR Vb CDR3 sequences from peripheral CD4+T cells isolated from 7 patients with PBC and healthy volunteers. All sequencing data were analyzed, together with corresponding clinical information, by bioinformatic software. The Mann-Whitney U test was used for statistical analysis.

RESULTSThe PBC patients showed a lower level of diversity among the peripheral CD4+TCR Vb CDR3 than the healthy volunteers, and patients with higher level progression of the disease showed a greater lack of diversity. In addition, 4 specific preferred-usage amino acid sequences were discovered for the PBC patients: ASSFTGGPVEQY, ASSLISSGNNEQF, ATSRDTLAGGPGDTQY, and SASLEGNTEAF; these sequences were also found in higher frequencies in patients with later stages of PBC.

CONCLUSIONSDecreased TCR Vb CDR3 diversities and specific preferred usage of TCR CDR3 sequences in peripheral CD4+T lymphocytes in PBC suggest that clonal expansion of a large number of CD4+T cells may be an important factor for PBC progression. These data provide a better understanding about the general characteristics of CD4+T cells in PBC patients and related to pathogenesis of the disease, and may provide useful insights into potential targets for immunotherapy.

Amino Acid Sequence ; CD4-Positive T-Lymphocytes ; High-Throughput Nucleotide Sequencing ; Humans ; Liver Cirrhosis, Biliary ; Polymerase Chain Reaction ; Receptors, Antigen, T-Cell

Objective: To clarify the situation of the detection of upper gastrointestinal cancer and precancerous lesions in the Feicheng city and discuss the possible influencing factors. Methods: A cluster sampling method was used to determine the participants. A unified questionnaire was used to investigate the basic information including history of alcohol intake, smoking and chinese tea, as well as other eating habits, medical history of digestive tract and cancer. Endoscopy was used to to screen the patients with upper gastrointestinal cancer and precancerous lesions. Influential factors were explored by non-conditional logistic regression model. Results: 911 of 7 291participants were positive, and the total detection rate was 12.49%. The total positive detection rate of male and female was 17.94% and 8.71%, respectively (P<0.001). Multivariate logistic regression analysis showed that the sex (OR=0.527, 95%CI: 0.440-0.631), age (OR=2.037, 95%CI: 1.849-2.245), smoking (OR=1.240, 95%CI: 1.014-1.516) and alcohol consumption (OR=1.232, 95%CI: 1.012-1.500) , meat and protein intake (OR=0.794, 95%CI: 0.638-0.987) and drink tea (OR=1.233, 95%CI: 1.056-1.440) may be influencing factors of the total detection rate of upper gastrointestinal cancer and precancerosis. Conclusions In Feicheng city, intake of meat and protein is the protective factor of the upper gastrointestinal cancer and precancerous lesions. Men, aged, smoking, drinking and tea will increase the risk of upper gastrointestinal cancer and precancerous lesions.