Malaysia first reported H5N1 poultry case in 2004 and subsequently outbreak in poultry population in 2007. Here, a recombinant gene encoding of peptide epitopes, consisting fragments of HA1, HA2 and a polybasic cleavage site of H5N1 strain Malaysia, was amplified and cloned into pET-47b(+) bacterial expression vector. DNA sequencing and alignment analysis confirmed that the gene had no alteration and in-frame to the vector. Then, Histagged truncated HA protein was expressed in Escherichia coli BL21 (DE3) under 1 mM IPTG induction. The protein expression was optimized under a time-course induction study and further purified using Ni-NTA agarose under reducing condition. Migration size of protein was detected at 15 kDa by Western blot using anti-His tag monoclonal antibody and demonstrated no discrepancy compared to its calculated molecular weight.

INTRODUCTIONUltrasound-guided fine-needle aspiration cytology (US-FNAC) of thyroid nodules is an important diagnostic procedure. In most hospitals, patients are referred to radiologists for US-FNAC, but this often results in a long waiting time before results are available. Surgeon-performed US-FNAC (SP-US-FNAC) during the initial patient consultation attempts to reduce the waiting time but it is not known whether this is as accurate as radiologist-performed US-FNAC (RP-US-FNAC). The aim of this study is to compare the clinical efficiency between SP-US-FNAC and RP-US-FNAC.

MATERIALS AND METHODSA retrospective study was performed on patients from the Department of General Surgery, Tan Tock Seng Hospital (TTSH) who underwent an US-FNAC from August 2011 to May 2012. All cases of SP-US-FNAC were performed by a single surgeon. This study compared the rates of positive diagnoses achieved by SP-US-FNAC and RPUS- FNAC as well as the time interval to reach a cytological diagnosis by each group.

RESULTSA total of 40 cases of SP-US-FNAC and 72 cases of RP-US-FNAC were included in the study. SP-US-FNAC resulted in 28 (70%) positive diagnoses and 12 (30%) nondiagnoses while RP-US-FNAC resulted in 47 (65.3%) positive diagnoses and 25 (34.7%) non-diagnoses. These results were comparable (P=0.678). The median time taken to reach a cytological diagnosis was 1 working day for SP-US-FNAC and 29.5 working days for RP-US-FNAC resulting in a shorter interval to reaching a cytological diagnosis for SP-US-FNAC (P<0.001).

CONCLUSIONIn the workup of thyroid nodules, SP-US-FNAC is as accurate as RP-US-FNAC but significantly reduces the time taken to reach a cytological diagnosis. This leads to greater clinical efficiency in the management of patients with thyroid nodules, which in turn leads to other benefits such as decreased patient anxiety and increased patient satisfaction.

Adult ; Biopsy, Fine-Needle ; methods ; Female ; Humans ; Image-Guided Biopsy ; Male ; Middle Aged ; Retrospective Studies ; Specialties, Surgical ; Thyroid Nodule ; pathology ; Time Factors

While perforations, postoperative fistulas, and leaks have traditionally led to surgical or interventional radiology consultation for management, the introduction of the over-the-scope clip has allowed increased therapeutic possibilities for endoscopists. While primarily limited to case reports and series, the over-the-scope clip successfully manages gastrointestinal bleeding, perforations, as well as postoperative leaks and fistulas. Retrospective studies have demonstrated a relatively high success rate and a low complication rate. Given the similarity to variceal banding equipment, the learning curve with the over-the-scope clip is rapid. However, given the higher risk of procedures involving the use of the over-the-scope clip, it is essential to obtain the scope in a stable position and grasp sufficient tissue with the cap using a grasping tool and/or suction. From our experience, while closure may be successful in lesions sized up to 3 cm, successful outcomes are obtained for lesions sized <1 cm. Ultimately, given the limited available data, prospective randomized trials are needed to better evaluate the utility of the over-the-scope clip in various clinical scenarios, including fistula and perforation management.

Objective To identify new recombinant antigens with potential for diagnosis of hemorrhagic fever with renal syndrom (HFRS) and establish reverse transcription-polymerase chain reaction-restriction fragments length polymorphism (RT-PCR-RFLP) for genotyping of hantavirus. Methods One group of primers was used to clone the full-length S genome segment and the partial S genorme segment of the N-terminal. The two cloned genes were both fusionally expressed and non fusionally expressed in the T7 system. The other group of primers was used to establish a RT-PCR method to detect RNAs in 37 virus isolates, 2 positive standard virus strains of hantavirus and 5 negative controls. The method for typing RFLP was set up by digesting the PCR products of 20 virus isolates with Ras Ⅰ and Hind Ⅲ. Results The non-fusionally expressed products with a working concentration of 1:10 000 by chapping enzyme-linked immunosorbent assay (cELISA), presented good biological activity though yields were lower than that of the fusionally expressed products.The specific component of the hantavirus genome (299 bp or 577 bp) wes seen in all viral samples. The genotyping of hantavirus showed that 9 out of the total were hantann (HTN) viruses, 8 were seol (SEO) viruses and 3 were not determined. Conclusions The good working titrer of expressed recombinant antigen showed that it has the potential to replace the natural antigen for detecting hantavirus antibodies. On comparison with cELISA, the detection rates for these two methods were 100% and 84.6%, and the coincidence rate was 84.6%. The former had a 15.4% higher sensitivity than the latter. The typing efficiency of RT-PCR-RFLP and sero-typing method was 85% (17/20) and 55% (11/20), respectively, showing that the former was 30% higher than the latter, while their results were highly consistant.

A healthy 30-year-old woman underwent health screening, and was incidentally found to have a well-defined echogenic lesion in Segment VI of her liver, measuring 4.2 X 1.8cm in her ultrasound scan. She was asymptomatic, has no cutaneous lesions, nor stigmata of chronic liver disease. She has no hepatomegaly. CT scan of her abdomen found multiple liver haemangiomas, varying in size from 21 X 28mm to sub-centimeter lesions. Diagnosis of hepatic haemangioma in this patient is based on typical appearance on contrast imaging. Their appearance is important to recognise as they do not need intervention and attempts at instrumentation for diagnosis or surgery may result in more harm than good.

Illegally paid blood donation was a risk factor for HIV acquisition exclusively in Henan and Hubei Provinces of China, and not in Shanghai. Nucleotide sequences in the gag and env genes of HIV-1 were compared between isolates from Henan and Shanghai regions of China to test whether an expected higher degree of a common source of infections from this unique blood donation transmission risk would be evident as decreased variation among Henan isolates in an exploratory cross-sectional analysis. Among 38 isolates studied, 23 of 23 (100%) from Henan and 8 of 15 (54%) from Shanghai were subtype B. In addition, fewer sequence differences were found in gp41 of subtype B isolates from Henan than from Shanghai isolates. Further studies with additional controls are therefore warranted to confirm the role of the degree of a common source of infections in differences in HIV variation across populations.

OBJECTIVETo assess the relationship between microglia activation and apoptosis of neurons, and the significance of activated microglias in the formation and progression of senile plaques in Alzheimer's disease.

METHODSIL-1alpha and beta-amyloid immunohistochemistry, combined with TUNEL assay were used to assess brain tissue samples from 10 patients with Alzheimer's disease and 4 negative control cases without neurological disease.

RESULTSThe number of resting microglias in the brains of Alzheimer's disease patients was similar to that of the control group (P > 0.05), but the number of activated microglias was significant greater in the Alzheimer's disease patients than that of the controls (P < 0.01). The activated microglias displayed altered size and morphology, and was therefore, categorized into three subtypes as primed, enlarged and phagocytic microglias. The numbers of primed, enlarged and phagocytic microglias were 5.4 +/- 0.87, 11.5 +/- 1.25, and 3.4 +/- 0.32 microglia/mm2 and represented 26.6%, 56.65%, and 16.75% of all activated microglias respectively. The number of TUNEL positive apoptotic neurons was significantly greater in Alzheimer patients than that in the control group (P < 0.05). There was a close relationship between the apoptosis of neurons and the activation of microglias (P < 0.01). The activated microglias were differentially distributed among four different plaque types in Alzheimer patients. Many primed (42.3%) and most of the enlarged and phagocytic microglias (56.2% and 70.6%) were present in the diffuse neuritic plaques.

CONCLUSIONSHyperplasia and activation of microglias are a common phenomena in AD and may play an important role in its pathogenesis. There is a close relationship between the apoptosis of neurons and activation of microglias. The activation of microglias may play a key pathogenic role in senile plaque formation and progression of Alzheimer disease.

Aged ; Alzheimer Disease ; etiology ; pathology ; Amyloid beta-Peptides ; analysis ; Apoptosis ; Cell Count ; Cell Differentiation ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Interleukin-1 ; analysis ; Microglia ; chemistry ; classification ; pathology ; Middle Aged ; Phagocytes ; pathology

OBJECTIVETo study the chemical constituents of the leaves of Naudea officinalis.

METHODThe chemical constituents were separated by column chromatography and semi-preparative HPLC techniques, and their structures were determined by spectral analysis.

RESULTFive compounds were isolated and identified as strictosamide (1), 10-hydroxy strictosamide (2), kaempferol-3-O-rutinoside (3), rutin (4), pumiloside(5).

CONCLUSIONAmong these compounds, 2, 3, 4 are isolated from the leaves of Naudea officinalis for the first time.

Camptothecin ; analogs & derivatives ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Kaempferols ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; methods ; Plant Extracts ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Rutin ; chemistry ; isolation & purification ; Vinca Alkaloids ; chemistry ; isolation & purification

OBJECTIVETo study on the purification of strictosamide from Nauclea officinalis by macroporous resin to provide reference for production.

METHODThe best macroporous resin was selected among 10 kinds of resins according to adsorption and desorption of the static adsorption experiments. The adsorption quantity, elution volume of water, concentration and elution volume of alcohol were determined according to the single factor experiment.

RESULTHPD400 was the best resin, and the best adsorption quantity was 20.23 mg x g(-1), the elution volume of water and 30% alcohol was 6 BV, and the elution volume of 70% alcohol was 4 BV.

CONCLUSIONThis technology is simple, feasible, and it can provide reference for the industrialized production.

Adsorption ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethanol ; chemistry ; Resins, Synthetic ; chemistry ; Rubiaceae ; chemistry ; Technology, Pharmaceutical ; methods ; Vinca Alkaloids ; chemistry ; isolation & purification

To explore the mechanism of needling Ying method for treatment of sore throat. By the analysis of pathogenesis of sore throat, the authors think the key of its pathogenesis is stagnation of pathogenic factors such as hotness and phlegm accumulating, and meridian-vessel obstruction in the throat is its meridian foundation. There are several meridians passing through the throat, so the throat is closely related to viscera and meridians, and stagnation of pathogenic factors such as hotness and phlegm accumulating in the throat lead to sore throat when exogenous pathogenic factors invading or dysfunction of viscera and meridians. The treatment of needling Ying at local throat or combined with corresponding meridian point selection can dredge collaterals, dispel pathogenic factors, remove pathogenic factors to dispel swelling, resolve phlegm and dissipate stagnation and harmonize yin and yang, so as to relieve sore throat. In conclusion, needling Ying method is an important method in the treatment of sore throat.
Acupuncture Therapy ; methods ; Bloodletting ; Humans ; Meridians ; Needles ; Pharyngitis ; etiology ; therapy ; Yin-Yang