Objective To develop a method for detecting the genotype of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) alleles. Methods According to the sequence characteristic of PvMSP-1, nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to amplify the polymorphic region of ICB5-ICB6 which contains Q repeats and PvuII restriction site (Sal-1 type). The PCR product was digested by PvuII restriction endonuclease and the digested fragments were observed by 2% agarose gel electrophoresis. The allelic type was determined according to the banding pattern. Results Bands in size of 400 bp (Belem type ) and/or 470 bp (Sal-1 type ) appeared in all 98 P. vivax isolates, no band was found in negative control. After PvuII digestion, two Sal-1 type fragments (120 bp and 350 bp) were obtained from 45 samples of 470 bp. Single-band of 400 bp appeared in 3 of 40 samples with 400 bp as Belem type, two bands of 120 bp and 280 bp appeared from other 35 samples as recombination type III, and another 2 bands with 120 bp and 240 bp as Korean isolate. Conclusion The result showed that the nested PCR-RFLP may be applied in the detection and identification of the three PvMSP-1 allelic types in China.

Objective To provide the basis for the differentiation with similar species of intestinal flukes through observing the figure of Haplorchis pumilio. Methods Adults of H. pumilio were collected from the intestine of the cat which was infected with the encysted cercariae of H.pumilio for 45 days. The worms were observed after staining. Eggs and metacercariae of H.pumilio were collected and examined for their shape, size and morphological characteristics. Pseudorasbora parva, the fish host, was examined for the parasitized sites of metacercariae. Results The principal characteristics of the adults is the acetabulum degradation. There are only the genital sucker with 44-48 hamuli. The average measurement of eggs is 31.2?16.7 ?m with a smooth shell. Its aceromion is not evident. The average diameter of metacercariae is 168.5 ?m. There are squamous spines on metacercaria. The metacercariae only parasitize in the muscle between the basis of the fin and the fish body. The average measurement of metacercaria cyst is 445?95?m, with squamous spines on the body surface. Hamuli are found on the genital sucker of metacercaria cyst. Conclusion The morphological figures and parasitic sites of metacercaria, the genital sucker of the adult, and the number and form of the hamulus on the genital sucker provide basis for distinguishing H. pumilio from other intestinal flukes.

Objective To develop methods of extracting DNA from malaria parasites on Giemsa-stained blood smears. Methods Improved Na2HPO4 method and Chelex-100 ion-exchange technique were used to extract DNA from Giemsa-stained or unstained blood smears. Nested PCR was employed for amplification and identification of allelotypes in the Plasmodium vivax merozoite surface protein-1(PvMSP-1). Results Target DNA bands appeared in all samples of unstained thick blood smears, while no DNA bands were visible in the fixed and stained thin smears. Both methods identified PvMSP-1 alleles from smears with parasitemia of ≥0.01%. Conclusion It is feasible to identify PvMSP-1 alleles from Giemsa-stained blood smear.

A simple, rapid, and sensitive gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the simultaneous determination of two fatty acids, methyl hexadecanoate (MH) and methyl stearate (MS), to allow the evaluation of packaging-drug compatibility. The two migrants were quantified in selective ion-monitoring (SIM) mode, with limits of detection (LOD) of 0.0030 μg/mL and 0.0121 μg/mL. Linear calibration curves for MH and MS were obtained in the concentration ranges of 0.1011–5.0570 μg/mL and 0.2015–10.0740 μg/mL, respectively. The developed method was successfully applied to estimate the safety of the injection of recombinant antitumor-antivirus protein (RAAP). The results showed that the possible maximum daily intake was 3.0 ng and 12.1 ng for MH and MS, re-spectively. As these values were both below the permitted daily exposure, the migrants can be con-sidered as having low safety risk and do not affect the quality of the injection.

A simple, rapid and sensitive method based on an ultra-performance liquid chromatography–tandem mass spectrometry(UPLC–MS/MS)has been developed and validated for the determination of pimavanserin in rat plasma.The analyte was extracted by protein precipitation with methanol and separated on an ACQUITY BEH C18column(100 mm×2.1 mm,1.7μm;Waters,USA),with an isocratic elution of acetonitrile-water containing 10 mM ammonium acetate (70:30, v/v), at a flow rate of 0.2 mL/min for 2.5 min. The analyte and clarithromycin (the internal standard) were detected and quantified in positive ion mode using multiple reaction monitoring transitions at m/z 428.2 → 223.0 for pimavanserin and m/z 748.5 → 589.5 for clarithromycin. Relative coefficient (r) for the calibration curve was more than 0.9980. The intra-day and inter-day precisions(relative standard deviation,RSD%)were less than 13.3% and 10.5%,respectively,and the accuracy(relative error,RE%)was within ± 11.5%.The analytical method was successfully applied to a routine pharmacokinetic study of pimavanserin in rats after oral administration at the dose of 10 mg/kg.

Objective To study the correlations of VDR FokⅠgene polymorphism with type 2 diabetes in postmenopausal women of Han nationality in south Sichuan. Methods 160 patients with type 2 diabetes (T2DM) and 190 healthy cases were enrolled in the study. The VDR FokⅠgene polymorphisms were detected using RFLP-PCR and DNA sequencing. Results The FF, Ff and ff genotype frequencies were 32.5%, 47.5%and 20% in the T2DM group and 15.8%, 53.7%, 30.5% in the control group, respectively (P < 0.05). The allele frequencies were 56.3%, 43.8% in the T2DM group and 42.6%, 57.4% in the control group, respectively (P < 0.05). The risk of T2DM in the FF genotype people was 2.568 times higher than Ff/ff genotype (adjusted OR = 2.568, 95%CI = 1.246 ~ 5.292, P < 0.05). The levels of 2 h PG and HbA1C in the FF genotype people were significantly higher than those of the Ff/ff genotype people (P<0.05). Conclusions There was an association between the VDR FokⅠgene polymorphism and type 2 diabetes incidence in the postmenopausal women in south Sichuan area.

Objective To analyze the characteristics of tuberculosis (TB) in renal-transplant recipients from our hospital, and summarize the corresponding experiences in diagnosis and management.Methods A retrospective study was performed on 61 documented post-transplant TB cases out of the 2842 patients who received kidney transplantation in the First Affiliated Hospital of Sun Yat-sen University between Jan.1991 and Dec.2010.Results TB in the post-renal-transplant population in our hospital displayed the following characteristics:(1) High incidence (2.1％ ).54.1％ recipients were diagnosed within the first year post-transplant; (2) Lung was the most common site (77.0 ％).There was high prevalence (60.7 ％) of extra-pulmonary TB (lymphatic TB,23.0 ％; pleuritis,13.1 ％; graft,11.5％); (3) Fever (83.6 ％),cough (55.7 ％),sputum (41.0 ％) were the most common clinical manifestations.There were also emaciation (3.3 ％) and enlargement of lymph nodes (18.0 ％); (4) Chest X-ray and CT were of great value during TB diagnosis while purified protein derivative of tuberculin (PPD) skin test had little diagnostic value with a negative result in 56 cases (91.8 ％) ; (5) Liver function damage ( 16.4 ％),kidney function injury (39.3 ％) and peripheral nerve toxicity (3.3 ％) were the main adverse reactions of anti-tuberculosis chemotherapy,also the major cause of anti-TB failure; (6) Pre-transplant TB (17 cases) increased the probability of TB recurrence (4 cases,23.5 ％) post-transplantation; (7) The post-transplant TB patients were accompanied with cellular immune deficiency,resulting in overlapping infection of bacteria,viruses and fungi (19.7 ％); (8) 1- and 3-year patient/graft survival rate of patients with post-transplant TB was 85.2 ％/78.7 ％ and 85.2 ％/75.4 ％ respectively. The accumulative mortality rate reached to 14.8％,while overlapping infection was the major cause of death (66.7 ％).Conclusion Chinese renal transplant recipients still face a high risk of TB because of their immunecompromised state and epidemiological prevalence of the disease. For the high mortality rate and associated serious complications,rapid diagnosis and effective anti-TB chemotherapy are of great value for TB population.

Carcinoma, Medullary ; metabolism ; Carcinoma, Neuroendocrine ; Humans ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NADP ; metabolism ; Neoplasm Proteins ; metabolism ; Prognosis ; Thyroid Neoplasms ; metabolism

Based upon the underlying mechanism and pathological evidence of tissue injury of antibody-mediated rejection (AMR) , four etiological and symptomatic therapies were proposed for managing AMR, including etiological treatment of AMR including antibody-targeting, B cell or plasma cell-targeting therapies; strategies for preventing antibody-mediated endothelial damage: an inhibition of complement/antibody dependent cell-mediated pathways; anticoagulant & thrombolytic therapies for thrombotic microangiopathy secondary to endothelial damage ; anti-inflammatory therapies for acute/chronic vascular inflammation secondary to endothelial damage. Etiological treatment is essential for preventing and treating AMR while symptomatic measures, such as anticoagulant, thrombolytic and antiinflammatory therapies, are stressed. Finally the authors devised therapeutic strategies for AMR in 4 different patient groups of non-sensitized allograft recipients, sensitized allograft recipients, individuals with active AMR and those with chronic active AMR.

Herein, we report a novel sensor to detect trypsin using a purpose-designed fluorescein-labelled peptide with negatively charged carbon nanoparticles (CNPs) modified by acid oxidation. The fluorescence of the fluorescein-labelled peptide was quenched by CNPs. The sensor reacted with trypsin to cleave the peptide, resulting in the release of the dye moiety and a substantial increase in fluorescence intensity, which was dose-and time-dependent, and trypsin could be quantified accordingly. Correspondingly, the biosensor has led to the development of a convenient and efficient fluorescent method to measure trypsin activity, with a detection limit of 0.7μg/mL. The method allows rapid determination of trypsin activity in the normal and acute pancreatitis range, suitable for point-of-care testing. Furthermore, the applicability of the method has been demonstrated by detecting trypsin in spiked urine samples.