BACKGROUNDNowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase II beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E. coli).

METHODSThe ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E. coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot.

RESULTSA 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase II beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E. coli JM109. The recombinant protein could be recognized by the S. japonicum infected rabbit serum.

CONCLUSIONThe full-length cDNA sequences encoding S. japonicum casein kinase II beta subunit were firstly sequenced, cloned, and expressed in E. coli.

Amino Acid Sequence ; Animals ; Base Sequence ; Blotting, Western ; Casein Kinase II ; chemistry ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; isolation & purification ; Escherichia coli ; genetics ; Molecular Sequence Data ; Rabbits ; Schistosoma japonicum ; enzymology ; genetics

OBJECTIVETo investigate the effect of ultrasonic wave on extracting Konjac Glucomannan(KGM) in Konjac refined powder.

METHODFree reduced sugar in Konjac refined powder was removed and Konjac refined powder in the aqueous solution was processed by ultrasonic wave and KGM content was measured by spectrophotometry.

RESULTKGM content in the Konjac refined powder aqueous solution by ultrasonic process at fixed 40 kHz, 100 W, 30-45 min was equal to that by routine method at 4 h; whereas, by 1 h of ultrasonic process, KGM content was significantly enhanced than that by 4 h of routine method(P < 0.01), enhancement rate was 6.5%. Linearity of standard glucose was good (r = 0.9996) in range of 0.2-1.6 mg. The average recovery was 97.8%, RSD of repeatability was 1.27%.

CONCLUSIONUltrasonic extraction in aqueous solution is a reliable and rapid method that can enhance extraction efficiency of KGM in Konjac refined powder.

Amorphophallus ; chemistry ; Mannans ; analysis ; isolation & purification ; Powders ; Ultrasonics

Angiostrongyliasis, caused by Angiostrongylus cantonensis infection, is a food-borne parasitic disease. Its larvae evoke eosinophilic inflammation in the central nervous system, but can also cause pathological changes in the eyes. Among ocular angiostrongyliasis cases, the incidence of optic neuritis is low and only few sporadic reports exist. Some patients with optic neuritis developed obvious hypopsia or even vision loss, which would seriously influence the quality of life of patients. Prompt treatment of optic neuritis caused by A. cantonensis is the key factor for minimizing the incidence of serious complications of this disease. In this review, we first provide a comprehensive overview of ocular angiostrongyliasis, and then focus on the clinical features of optic neuritis caused by A. cantonensis.
Angiostrongylus cantonensis/*isolation & purification ; Animals ; Asia/epidemiology ; Eye Diseases/*epidemiology/*parasitology/pathology ; Humans ; Incidence ; Optic Neuritis/*epidemiology/*parasitology/pathology ; Strongylida Infections/*epidemiology/*parasitology/pathology

OBJECTIVETo study the expression of plasma miRNA-497 in children with sepsis-induced myocardial injury and its clinical significance.

METHODSA total of 148 children with sepsis were enrolled. According to the presence or absence of myocardial injury, these children were divided into myocardial injury group (n=58) and non-myocardial injury group (n=90). The two groups were compared in terms of the changes in plasma levels of miRNA-497, cardiac troponin I (cTnI), creatine kinase-MB (CK-MB), N-terminal pro-brain natriuretic peptide (NT-proBNP), procalcitonin (PCT), and C-reactive protein (CRP) and left ventricular ejection fraction (LVEF). The receiver operating characteristic (ROC) curve was plotted to evaluate the value of plasma miRNA-497, cTnI, and CK-MB in the diagnosis of myocardial injury. A Pearson correlation analysis was used to determine the correlation of miRNA-497 with cTnI, CK-MB, NT-proBNP, PCT, CRP, and LVEF.

RESULTSCompared with the non-myocardial injury group, the myocardial injury group had significantly higher plasma levels of miRNA-497, cTnI, CK-MB, NT-proBNP, PCT, and CRP (P<0.05). Plasma miRNA-497, cTnI, and CK-MB when measured alone or in combination had an area under the ROC curve of 0.918, 0.931, 0.775, and 0.940 respectively. At the optimal cut-off value of 2.05, miRNA-497 had a sensitivity of 90.4% and a specificity of 91.2%. The correlation analysis showed that there was a good correlation between plasma miRNA-497 and cTnI in children with myocardial injury (r=0.728, P<0.01).

CONCLUSIONSPlasma miRNA-497 has a similar value as cTnI in the diagnosis of sepsis-induced myocardial injury in children and may be used as a potential marker for early diagnosis of myocardial injury.

OBJECTIVETo Summarize the clinical experience of individual immunosuppressive regime in heart transplantation with high risk.

METHODSFrom September 2001 to December 2006, 51 cases with the complication of Hepatitis B viruses (HBV) infection, diabetes mellitus, renal dysfunction or pulmonary infection in perioperative period were analyzed retrospectively. All cases received daclizumab (Zenapax) induction therapy, and baseline triple immunosuppressive regime was consist of cyclosporine (CsA), azathioprine (Aza) or mycophenolate mofetil (MMF) and prednisone (Pred). Ten cases received HBV infection in preoperative period, the immunosuppressive protocol was emphasized on the use of MMF and the withdraw of Pred one month later in postoperation. Nine cases received diabetes mellitus in pre-operation, 4 cases had post-transplant diabetes mellitus. The immunosuppressive protocol was emphasized on the use of CsA rather than FK506, the use of Pred was less dosage, and the therapy of insulin was necessary. Sixteen cases had renal dysfunction in pre-operation, the use of MMF was routine but the use of CsA was delayed to the time 5 to 19 d postoperative. Twelve cases received pulmonary infection after allograft transplantation. The immunosuppressive agent was to be taped or suspended in therapy time.

RESULTSThe liver function of the 10 cases with HBV infection was stable in 1 year follow-up, and 1 case received acute rejection after 13 months allograft transplantation. In the 6 months follow-up, the blood glucose level of the 13 cases with diabetes mellitus was stable, none of the cases suffered from acute rejection. In the one month follow-up, none of the 16 cases with renal dysfunction suffered from acute rejection, and the renal function was normal. Two of the 12 cases with the pulmonary infection were died of serious infection, others were survival. One case received acute rejection on the 17th day in postoperation.

CONCLUSIONSLow mortality can be realized by selecting appropriately individual immunosuppressive regime and the episode of acute rejection is rare.

Adult ; Female ; Follow-Up Studies ; Graft Rejection ; prevention & control ; Heart Transplantation ; Humans ; Immunosuppressive Agents ; administration & dosage ; therapeutic use ; Male ; Perioperative Care ; Retrospective Studies ; Risk Factors

Dengue ; epidemiology ; Dengue Virus ; genetics ; isolation & purification ; Evolution, Molecular ; Genes, Viral ; Humans ; Viral Envelope Proteins ; genetics

To optimize the belt drying process conditions optimization of Gardeniae Fructus extract from Reduning injection by Box-Behnken design-response surface methodology, on the basis of single factor experiment, a three-factor and three-level Box-Behnken experimental design was employed to optimize the drying technology of Gardeniae Fructus extract from Reduning injection. With drying temperature, drying time, feeding speed as independent variables and the content of geniposide as dependent variable, the experimental data were fitted to a second order polynomial equation, establishing the mathematical relationship between the content of geniposide and respective variables. With the experimental data analyzed by Design-Expert 8. 0. 6, the optimal drying parameter was as follows: the drying temperature was 98.5 degrees C , the drying time was 89 min, the feeding speed was 99.8 r x min(-1). Three verification experiments were taked under this technology and the measured average content of geniposide was 564. 108 mg x g(-1), which was close to the model prediction: 563. 307 mg x g(-1). According to the verification test, the Gardeniae Fructus belt drying process is steady and feasible. So single factor experiments combined with response surface method (RSM) could be used to optimize the drying technology of Reduning injection Gardenia extract.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Gardenia ; chemistry ; Research Design ; Vacuum

Schistosomiasis is an important zoonotic parasitic disease, and is categorized as a neglected tropical disease by the World Health Organization. Following the concerted efforts for more than 70 years, great achievements have been made in the national schistosomiasis control program in China, and the prevalence, disability and mortality due to schistosomiasis has remarkably dropped. Nevertheless, the frequent identification of imported schistosomiasis and the resulting potential transmission risk in mainland China have been recently paid much attention following the implementation of the “Belt and Road Initiative” and the China-Africa Cooperation Forum. This review describes the advances in the diagnostic tools for schistosomiasis, including pathogenic techniques, immunodiagnostic techniques and nucleic acid assays, in order to consolidate schistosomiasis control achievements and promote the capability for detection of external biological safety risks.

OBJECTIVETo establish a quantitative method for the content determination and monosaccharide composition analysis of Konjac glucomannan (KGM) in Konjac refined powder by pre-column derivatization high performance liquid chromatographic method (HPLC).

METHODThe two derivatives combined reducing monosaccharides with 1-phenyl-3-methyl-5-pyrazolone (PMP) were separated by reverse-phase HPLC using a developed fragment gradient elution process, and monitored by ultraviolet detector at 250 nm. The broad reagent peak of PMP was separated very well from all the PMP-sugars, and good separation was achieved for derivatives of mannose and glucose. The quantitative methods of two reducing monosaccharides were studied by the method combined internal and external standard; while the KGM content in Konjac refined powder was determined.

RESULTLinearity of glucose was good (r = 0.9990) in range of 1.002-8.016 nmol; while mannose (r = 0.9994) in range of 1.001-8.008 nmol. The average recovery of this method was 98.1%, RSD of repeatability was 1.72%. KGM content in Konjac refined powder was 79.5%, ratio of glucose to mannose in KGM was 1:1.51.

CONCLUSIONThis method is a sample, convenient and rapid method that can determine KGM content and analyze monosaccharide compositions in KGM, which will be helpful to quality assessment of Konjac refined powder.

Amorphophallus ; chemistry ; Chromatography, High Pressure Liquid ; Glucose ; chemistry ; Mannans ; analysis ; chemistry ; Mannose ; chemistry ; Monosaccharides ; analysis ; Plants, Medicinal ; chemistry ; Powders ; chemistry

OBJECTIVEThe aim of this paper was to investigate the factors associated with viral response and HBeAg seroconversion and the relationship between them at different stages of interferon treatment in HBeAg-positive chronic hepatitis B patients.

METHODSPEG-IFN alfa-2a was injected subcutaneously in doses of 180 microg once a week for 48 weeks to HBeAg-positive chronic hepatitis B patients, and the patients were followed for another 24 weeks after the treatment. The serum HBV DNA load was measured by real-time quantitative PCR assay. Microparticle enzyme immunoassay analysis (MEIA) was then carried out by an automatic enzyme immunoassay analysis instrument to measure HBeAg and anti-HBe. Virological response and HBeAg seroconversion rates, and the factors associated with them were analyzed.

RESULTSThe differences in ALT baselines between viral responding and non-responding groups were significant at treatment time and at the end of the follow-up period. These differences were also significant in patients with HBeAg seroconversion at 12 weeks and at the end of the follow-up period compared with the non-conversion group. No significant difference of HBV DNA baseline was observed between the HBeAg seroconversion and non-conversion group. At 12, 24 and 48 weeks, in patients with viral response during the treatment, their HBeAg seroconversion rates were 43.8%, 21.4% and 18.9% respectively; their respective HBeAg seroconversion rates remaining at 72 weeks were 42.9%, 33.3% and 27.6%. HBeAg seroconversion was related to HBV DNA negativity at 48 weeks treatment in the multivariate analysis (OR=2.15, 95.0% CI=1.744-2.664, P less than 0.01).

CONCLUSIONSViral response and early and sustained HBeAg seroconversion were associated with pretreatment ALT levels. HBeAg seroconversion was related to viral response during IFN treatment, but not to the baseline HBV DNA load.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; Young Adult