OBJECTIVETo develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria.

METHODSType-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products.

RESULTSTwenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification.

CONCLUSIONThe novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.

Bacteria ; classification ; genetics ; isolation & purification ; DNA, Bacterial ; analysis ; Food Contamination ; analysis ; Food Microbiology ; methods ; Foodborne Diseases ; microbiology ; Humans ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.

METHODSTwo pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.

RESULTSThe RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.

CONCLUSIONThis multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza B virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors ; Viral Matrix Proteins ; genetics ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo discuss the mechanisms of electroacupuncture at "Neiguan" (PC 6) on p38 MAPK signaling pathway in rats with cardiac hypertrophy.

METHODSForty SD rats were randomly divided into four groups: a normal group, a model group, a model plus p38 MAPK inhibitor group, a model plus electroacupuncture group, ten rats in each group. The model rats were established by subcutaneous injection 3 mg/(kg x d) of Isoprenaline Hydrochloride; model plus p38 MAPK inhibitor group were injected 0.3 mg/(kg x d) of specific inhibitor SB 203580; model plus electroacupuncture group was treated by electroacupuncture at "Neiguan"(PC 6) with continuous-wave, 2 Hz and 1 mA for 20 minutes, once a day for 14 days. There was no treatment in other two groups. The contents of TNF-alpha and IL-1beta in heart tissue were detected by radioimmunoassay and the p38 MAPK, p-p38 MAPK by western blot.

RESULTSCompared with normal group, the contents of TNF-alpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly increased in model group (all P < 0.01). The contents of TNFalpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly decreased in model plus p38 MAPK inhibitor group and model plus electroacupuncture group, compared with model group, all P < 0.05; compared with normal group, P < 0.05, P < 0.01; but no significant difference between model plus p38 MAPK inhibitor group and model plus electroacupuncture group (P > 0.05).

CONCLUSIONElectroacupuncture at "Neiguan" (PC 6) can prevent the phosphorylation of p38 MAPK of myocardial hypertrophy, and the mechanism maybe adjust p38 MAPK signaling pathway by inhibiting the expression of TNF-alpha and IL-1beta.

Acupuncture Points ; Animals ; Cardiomegaly ; metabolism ; therapy ; Electroacupuncture ; Interleukin-1beta ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.

METHODSPrimers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.

RESULTSThe 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.

CONCLUSIONThe established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.

Astroviridae ; genetics ; isolation & purification ; Diarrhea ; virology ; Feces ; virology ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Norovirus ; genetics ; isolation & purification ; RNA, Viral ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rotavirus ; genetics ; isolation & purification ; Sapovirus ; genetics ; isolation & purification

Objective To compare the effect of the domestic infantile type video intubationscope (VIS) and stethoscope in positioning of double-lumen endobronchial tube (DLT). Methods 100 cases of patients underwent elective thoracic surgery requiring single lung ventilation were randomly divided into two groups: domestic infantile type video intubationscope group (group V) and stethoscope group (group S), with 50 cases in each. After intubating with a DLT, the positions of DLT were judged and adjusted by VIS (group V) and stethoscope (group S) respectively, and then reviewed by fiberoptic bronchoscopy (FOB), the positioning time and accuracy were recorded. Results Comparing with the group S, the positioning time of DLT was significantly shorter and the total positioning accuracy of DLT was significantly higher in group V (P < 0.05). Conclusion It is easy and quickly, high accuracy with domestic infantile type video intubationscope in positioning of DLT, which is worthy of clinical popularization and application.

Objective To compare the clinical effect of nasal intubation with domestic video intubationscope and Macintosh direct laryngoscope. Methods One hundred patients scheduled for oro-maxillo-facial operation, American Society of Anesthesiologists (ASA) grade I or II, aged 19 ~ 67 years, were randomly divided into the domestic video intubationscope group (group V) and the Macintosh direct laryngoscope group (group M), with 50 cases in each group. Nasal intubation was respectively performed with domestic video intubationscope (Group V) and Macintosh direct laryngoscope (group M). Cormark-Lehane grade (C-L classification), tracheal intubation time, first-time intubation success rate and tracheal intubation complications were recorded.Mean arterial pressure (MAP) and heart rate (HR) of before induction of anesthesia (T0), after induction of anesthesia (T1), at glottic exposure (T2), at intubation (T3), 1 min after intubation (T4) and 3 min after intubation (T5) were recorded. Results C-L classification in group V was significantly lower than that in group M (P < 0.05), intubation time in group V was significantly shorter than that in group M (P < 0.05), first-time intubation success rate in group V was significantly higher than that in group M (P < 0.05). Compared with T1, MAP was significantly higher at T3~ T4and HR was significantly faster at T3in group V, MAP was significantly higher at T2~ T4and HR was significantly faster at T2~ T3in group M (P < 0.05). Compared with group M, MAP and HR in group V were significantly lower at T2~ T4 (P < 0.05). The incidence of tracheal intubation complications in group V was significantly lower than that in group M (P < 0.05). Conclusions Compared with Macintosh direct laryngoscope,domestic video intubationscope in nasal intubation is better in glottic exposure,it could shorten tracheal intubation time ,increase first-time intubation success rate, maintain stable hemodynamics, has fewer tracheal intubation complications and is worthy of clinical application.

Objective?To compare the clinical effects of domestic video intubationscope (VIS) versus fiberoptic bronchoscope (FOB) for difficult nasotracheal intubation.?Methods?60 ASA Ⅰ or Ⅱ elective patients with difficult airway, Mallampati class Ⅲ or Ⅳ, aged 22 ～ 68 years, weight 53 ～ 82 kg, were randomly divided into domestic video intubationscope group (group V) and fiber bronchoscope group (group F), 30 cases in each group. The nasotracheal intubation was respectively guided by VIS in group V and FOB in group F. Tracheal intubation time, success rate of tracheal intubation and complications of tracheal intubation were recorded. MAP, HR and SpO2 were recorded at before anesthesia induction (T0), after anesthesia induction (T1), at glottic exposure (T2), at intubation (T3).?Results?Compared with T0, MAP and HR were significantly decreased in both groups at T1(P < 0.05). MAP and HR were significantly increased in both groups at T3 than those at T1(P < 0.05), and there were no significant differences between the two groups (P > 0.05). SpO2 during tracheal intubation was no significantly reduced in both groups. The tracheal intubation time were respectively (76.0 ± 18.0) s and (80.0 ± 20.0) s in group V and group F, and the one-time success rate of intubation in group V and group F were respectively 96.7% and 93.3%, but there were no significant differences between the two groups (P > 0.05). There was no significant difference in the incidence of tracheal intubation complications between the two groups (P > 0.05).?Conclusions?Compared with FOB, difficult nasotracheal intubation guided by domestic VIS also is a safe and reliable, fast and effective method with high intubation success rate and less complications of tracheal intubation.

Aim To determine the effect of cornel iri-doid glycoside ( CIG ) on human hepatocyte cell line (L-02) injured by D-galactosamine (GalN) and tumor necrosis factor-α( TNF-α) .Methods Firstly, CIG was extracted , separated and purified . Cell lesion model injured by D-GalN/TNF-αwas tested by MTT method.T-AOC, SOD, MDA and calcium ion concen-tration were taken as indicators to study the effects of CIG on L-02 cell injured by D-GalN/TNF-α.The ex-pression of p-PERK, p-eIF-2α, caspase-3 protein were detected by Western blot .Results 44 mg · L-1 D-GalN and 100 μg · L-1 TNF-αwere suitable for L-02 cell lesion model.CIG high, middle, low concentra-tion group could significantly increase the L-02 cell ac-tivity by 21％, 13％, 8％, respectively and SOD activity and T-AOC ability as well compared with model group.At the same time, they markedly reduced the MDA activity except the low concentration .Three con-centrations of CIG could reduce the expression of endo-plasmic reticulum stress related protein PERK , eIF-2αand apoptosis-associated protein caspase-3. Conclu-sions CIG could protect L-02 cells injured by D-GalN/TNF-α.Increasing the cellular antioxidant abili-ty, reducing the damage of endoplasmic reticulum stress and the expression of apoptosis-associated protein may be the possible mechanism .

OBJECTIVE: To investigate the therapeutic effects and mechanisms of Bianyanning on acute pharyngitis in rats, and to provide evidence and experimental data for its clinical application. METHODS: The acute pharyngitis of rats was induced by spraying ammonia directly to their throat. The model rats were randomly divided into model control group, the high-, medium- and low-dose group of Bianyanning, while normal rats were used as control group, 10 in each group. After the corresponding drug treatment, the symptoms and manifestations of each group were observed and recorded; 24 hours after last gavaging, blood samples of each group were collected from the abdominal aorta. The serum contents of interleukin 1-beta (IL-1β) and tumor necrosis factor alpha (TNF-α) were detected by ELISA. HE method was used to observe the characteristic of the lung tissues and the transmission electron microscopy method was used to observe the trachea cilia. RESULTS: After the treatment, compared with the model control group, the high-, medium- and low-dose group of Bianyanning, the symptoms of acute pharyngitis such as inflamed and congestive throat were relieved obviously. The morphological changes of lung and bronchus tissues were apparently improved. The contents of IL-1β and TNF-α in serum were decreased significantly. CONCLUSION Compound Bianyanning can promote the recovering process of acute pharyngitis, improve the morphology of lungs and bronchus, which may be related to inhibiting the releasing of the IL-1β and TNF-α in serum.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Interleukin-1beta ; metabolism ; Pharyngitis ; drug therapy ; immunology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

In order to clarify the chemical constituents of Cistanche deserticola cultured in Tarim desert, a systematically phytochemical investigation was carried out. The chemical constituents were isolated by column chromatography, such as silica gel, Sephadex LH-20, MCI gel, ODS and semi-preparative HPLC, and their structures were determined on the basis of MS, NMR spectroscopic analysis, and comparison with literature data. Four compounds were isolated from the 85% ethanol extract of the stems of C. cultured in Tarim desert. Their structures were identified as cis-tubuloside (1), cis-cistanoside (2), cis-cistanoside J (3), and cis-isocistanoside C(4). Compounds 1-4 were four new cis-phenylethanoid glycosides. Herein, we firstly report the ¹H, ¹³C-NMR data of the new compounds(1-4) for the first time. This study will provide the scientific evidence for comprehensively analyzing the chemical constituents of C. deserticola cultured in Tarim desert.