DNA, Circular ; isolation & purification ; DNA, Viral ; isolation & purification ; Hepatitis B ; diagnosis ; Hepatitis B virus ; genetics ; Humans

Objective To evaluate the fludarabine instead of cyclophosphamide in modified busulfancyclophosphamide (mBuCy) regimen as a new myeloablative conditioning regimen for the treatment of acute leukemia patients receiving allogeneic hematopoietic stem cell transplantation (HSCT).Methods The clinic data of 45 acute leukemia patients undergoing allogeneic HSCT were analyzed.Among them,23 patients received mBuCy as conditioning regimen and 22 patients received BuFlu regimen (fludarabine 40 mg·m-2·d-1 for 5 days,instead of cyclophosphamide in mBuCy).Hematopietic engraftment,regimen-related toxicity (RRT),graft-versus-host disease (GVHD),infection condition,non relapse mortality,and overall survival were compared between the two groups.Results All patients achieved hematopoietic reconstitution and complete donor chimerism except for one patient of mBuCy group died of cerebral hemorrhage during conditioning.The incidence of RRT was no significant differences (P ＞ 0.05).In BuFlu group,the incidence of virus infection was higher (P =0.009),and the incidence of Ⅲll-Ⅳ aGVHD were 26.l ％ (6/23) and 4.5 ％ (1/22) (P =0.046) in mBuCy and in BuFlu group respectively.With a median follow up of 41 months,the incidence of non relapse mortality in mBuCy group was 17.4 ％ (4/23) and in BuFlu group was 9.1％ (2/22) (P =0.665).In mBuCy group and in BuFlu group,the relapse rates were 30.3 ％ (7/23) and 40.9 ％ (9/22) (P =0.474),the 5-year overall survival rates were (55.1±11.9) ％ and (61.4±10.8) ％ (P =0.659),and disease-free survival rates were (44.5±12.1) ％ and (22.1±12.3) ％ (P =0.747),respectively.Conclusions Fludarabine instead of cyclophosphamide in mBuCy regimen as a new myeloablative conditioning regimen has well tolerance,lower incidence of sever GVHD,satisfied overall survival,but the risk of infection and replase should be considered.

Adenine ; adverse effects ; analogs & derivatives ; therapeutic use ; Adult ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; drug therapy ; Humans ; Infant ; Infant, Newborn ; Male ; Organophosphonates ; adverse effects ; therapeutic use ; Pregnancy ; Pregnancy Trimesters

Glutamate decarboxylase (GAD) can catalyze the decarboxylation of glutamate into γ-aminobutyrate (GABA) and is the only enzyme of GABA biosynthesis. Improving GAD activity and thermostability will be helpful for the highly efficient biosynthesis of GABA. According to the Ramachandran plot information of GAD 1407 three-dimensional structure from Lactobacillus brevis CGMCC No. 1306, we identified the unstable site K413 as the mutation target, constructed the mutant GAD by site-directed mutagenesis and measured the thermostability and activity of the wide type and mutant GAD. Mutant K413A led to a remarkably slower inactivation rate, and its half-life at 50 °C reached 105 min which was 2.1-fold higher than the wild type GAD1407. Moreover, mutant K413I exhibited 1.6-fold higher activity in comparison with the wide type GAD1407, although it had little improvement in thermostability of GAD. Ramachandran plot can be considered as a potential approach to increase GAD thermostability and activity.
Glutamate Decarboxylase ; metabolism ; Half-Life ; Industrial Microbiology ; Lactobacillus brevis ; enzymology ; Mutagenesis, Site-Directed ; Mutation ; Temperature

OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.

METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.

RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.

CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.

Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea

OBJECTIVETo investigate the spontaneous YMDD mutation rate.

METHODSSerum samples collected from 196 untreated chronic HBV patients were detected by primer-specific real-time PCR.

RESULTSAmong 196 patients, spontaneous YMDD variants were detected in 21 subjects (20 YVDD mutants and 1 YIDD mutant). YMDD variants account for more than 50%, 25% to 50%, 9% to 25% of total virus load in 1, 5 and 15 patients, respectively. Gender, age, HBeAg status, serum viral load, the state of disease and duration of infection were not associated with spontaneous YMDD mutation. Genotype B had higher spontaneous YMDD rate than genotype C (20.00% vs 7.38%, x(2) = 6.28, P < 0.05).

CONCLUSIONSpontaneous YMDD variants exist in chronic hepatitis B patients, Genotype B is associated with higher spontaneous YMDD rate.

Adolescent ; Adult ; Aged ; DNA Primers ; DNA, Viral ; analysis ; blood ; genetics ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Viral Load ; Young Adult

Objective To investigate the pathogenic and clinical features of bloodstream infections in patients with hematological malignancies for improving clinical treatment.Methods A total of 92 patients with hematological malignancy and positive blood culture treated during the period from September 2011 to September 2016 were analyzed,including clinical manifestations,treatment and prognosis.The distribution and antibiotic resistance of pathogens were also investigated.Results Of the 92 patients with bloodstream infection,64.1％ had underlying agranulocytosis.All patients had fever.Septic shock was found in 45.7％ cases.Elevated procalcitonin was detected in 82.6％ cases.The 107 isolates from blood stream included 75 (70.1％) strains of gram negative bacteria,27 (25.2％) strains of gram positive bacteria,and 5 (4.7％) strains of fungi.Escherichia coli showed higher resistance rate to ceftriaxone (80.9％) and levofloxacin (91.3％).Klebsiella pneumoniae (31.2％) and Acinetobacter baumannii (50.0％) strains showed relatively high resistance to imipenem.Gram positive bacteria were still sensitive to vancomycin and linezolid.Overall,35 (38.0％) patients died.The initial empirical treatment regimen had significant impact on patient outcome (P＜0.05).The mortality rate of initial carbapenem-based empirical treatment was slightly lower (28.6％ vs 46.2％) than that of non-carbapenem-based initial regimen,but the difference was not significant (P=0.163).Conclusions The outcome is poor in patients with hematological malignancy complicated with bloodstream infection.The main pathogen is gram negative bacteria in such infections,associated with high antibiotic resistance.The emergence of carbapenem resistance is an issue of concern.The effectiveness of initial empirical therapy may have significant effect on patient outcome.

Objective To investigate expression and significance of decorin(DCN)in liver tissue and serum of liver transplant patients with chronic rejection(CR).Methods Immunohistochemistry(SP method)was used to detect expression of DCN in liver tissue of 16 normal controls, 20 patients with cirrhosis, 46 liver translantion patients without CR and 8 patients with CR.Enzyme-linked immunosorbent assay method(ELISA method)was used to determined the content of DCN in serum of all research subjects.Results The expression of DCN was negative in normal hepatic tissues and with/without CR, cirrhosis tissues showed strong expression of DCN.The positive expression rate and the average optical density value of DCN in liver transplant tissues with CR had significant difference comparing with Cirrhosis tissues(25% vs 55%, 0.1249 ±0.0039 vs 0.2357 ±0.0396, P <0.01,while no statistic siqnificance compared to normal liver tissues and those without CR.The level of DCN in serum was significantly higher in liver transplant patients with CR, with significant difference comparing with normal people, liver cirrhosis and transplant liver patients without CR(54.0833 ± 6.0325)μg/L vs(1.0232 ± 0.9105)μg/L,(12.6202 ± 1.5370)μg/L,(17.7102 ± 2.3562)μg/L, P < 0.01).The concentration of DCN in serum showed a positive correlation with the degree of CR.Conclusions DCN showed negative expression in liver tissue and increased significantly in serum of liver transplantation patients with CR.This suggests that DCN may be involved in occurrence and development of CR.At the same time the determination of DCN in serum maybe become an important indicator of the early diagnosis, development and prognosis of CR for liver transplant patients.

ObjectiveTo study the Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL)of the elderly and explore its clinical characteristics. MethodsThe clinical manifestation,laboratory examination and treatment of two cases of EBV positive DLBCL of elderly were lescribed.Results EBV positive DLBCL of elderly onset was often between 70-79 years,usually presented with lymphadenopathy and may be extranodal,may present with massive splenomegaly,secondary immune hemolytic anemia and often had B symptom.Pathologically,this disease was characterized by a proliferation of atypical large B cells with rich reactive cells,especially T cell.ConclusionImmunohistochemically,the tumor cells of EBV positive DLBCL of elderly present with CD20 or CD79a,EBV-encoded small RNA (EBER) positive.This disease has aggressive clinical course,a poor response to standard treatment.Rituximab may be effective in a short periods,but the long effect is limited.Overall survival is short.The cause of death is mainly respiratory failure due to factor infection.

Antiviral Agents ; therapeutic use ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Male ; Middle Aged