Objective To diagnose the first imported case of Plasmodium ovale infection by laboratory detection. Meth?ods The epidemiological data and blood samples of the case were collected,and the samples were detected by the microscopic examination,rapid diagnostic test(RDT)and nested PCR. Results The patient was a construction worker backing from Con?go,Africa. He experienced the symptoms of irregular fever and weakness one month after returning in Lingyang Town,Junxian County. The results of RDT only suggested no?Plasmodium falciparum infection. Under the microscope,it was seen that the in?fected RBC were obviously disfigured and in irregular shape,the ring forms were thick and big,and also thick granulas in big trophozoite stage and schizont stage were found. The results of PCR showed that the size of amplified product was about 800 bp, which was conformed to that of P. ovale. Conclusion Though microscopic examination is the golden standard for malaria diag?nosis,as P. ovale is difficult to be identified under microscope,the microscopic method combined with PCR test can be used for definite diagnosis.

Objective To assess the correlation between the shear wave velocity(SWV) and the ratio of type Ⅰ /type Ⅲ collagen fibers in plaques using virtual touch tissue imaging quantification(VTIQ).Methods Fifty four patients (54 plaques) who scheduled to carotid endarterectomy(CEA) were enrolled in this study.Their carotid plaques were preoperatively examined using VTIQ.The value of SWE of each plaque was obtained.The SWV of the plaque was evaluated by VTIQ.The endarterectomy specimens were transferred to the pathological department immediately after surgery to evaluate the plaque types and the distribution of collagen fibers in plaques.According to HE staining,the 54 plaques were divided into stable and unstable group.Results Among 54 plaques,14 were stable plaque and 40 were unstable plaques.The thickness and stenosis rate had no significant difference between stable group and unstable group (both P ＞ 0.05).The mean value of SWV and the ratio of type Ⅰ/typeⅢ collagen fiber in stable plaques were higher than those in the unstable plaques (both P ＜0.05).Based on ROC analysis,a cut-off value of SWE of 3.725 m/s (AUC=0.936) showed a sensitivity of 92.9％,a specificity of 80.0％ and an accuracy of 83.3％ for predicting stable plaques (95％ CI 0.818-0.982,P ＜0.05).The value of SWVs was closely correlated to the ratio of type Ⅰ/typeⅢ collagen fiber in plaques(r =0.882,P ＜0.05).Conclusions SWV of the carotid atherosclerotic plaque can be used to reflect the ratio of the type Ⅰ/typeⅢ collagen fiber in the plaques,and to evaluate the stability of plaque.

To investigate the effects of Tongxinluo capsule on sciatic nerve apoptosis in spontaneous type II diabetic KK/Upj-Ay mice, in order to explore its mechanism for improving diabetic peripheral neuropathy (DPN). KK/Upj-Ay mice were selected as the DPN animal model and randomly divided into the model, Tongxinluo low, middle and high group (1, 2, 4 g x kg(-1)). C57BL/6 mice were selected as the control group. Mice were given intragastrically for 12 weeks. Paw withdrawal latency, motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) were detected. Apoptotic rate were detected by FCM. Bcl-2, Bax, Caspase-3 mRNA and protein expression in sciatic nerve were examined by Real-time PCR and Western blot. p38MAPK, p-p38MAPK expression were examined by Western blot. In this study,the authors found that Tongxinluo capsule could increase paw withdrawal latency, MNCV and SNCV. Apoptotic rate of sciatic, the expression of Bax and caspase-3 were lower, while Bcl-2 expression was higher in Tongxinluo group than those in model mice. The expression of p-p38MAPK significantly decreased in Tongxinluo group. The results showed that Tongxinluo capsule has protective effects on diabetic peripheral neuropathy of mice via inhibiting cell apoptosis and suppressing the expression of p-p38MAPK.
Animals ; Apoptosis ; drug effects ; Capsules ; administration & dosage ; Diabetic Neuropathies ; drug therapy ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Sciatic Nerve ; cytology ; drug effects

This study aimed to investigate the drug susceptibility of wild-type and mutant avian influenza A (H7N9) virus neuraminidase (NA) to oseltamivir and zanamivir. Codon optimized DNA of H7N9 (A/ Hangzhou/1/2013) NA was synthesized and constructed into the pcDNA3.1/His vector (NA(H7N9-WT)). Mutant NA(H7N9-H274Y) and NA(H7N9-R292K) plasmids were constructed by directed mutagenesis PCR using NA(H7N9-WT) plasmid as the template followed by sequencing. NA plasmids were transfected into 293T cells and cell lysates containing NAs were collected 48 h post-transfection. Wild-type and mutant NAs were analyzed by Western blotting and their activities were tested by the 4-MUNANA-based assay. All three NAs were expressed and enzymatic activities were confirmed. The effects of oseltamivir and zanamivir on all three NAs were then tested. It showed that the half maximal inhibitory concentrations (IC50s) of oseltamivir carboxylate on NA(H7N9-WT), NA(H7N9-H274Y) and NA(H7N9-R292K) were 1.6 nM, 15.1 nM, and > 1 000 nM with fold changes of 9 and > 625, respectively. The IC50 values of zanamivir on NA(H7N9-WT), NA(H7N9-H274Y), and NA(H7N9-R292K) were 1.1 nM, 1.4 nM, and 38.0 nM with fold changes of 1.3 and 34, respectively. These results indicated that oseltamivir and zanamivir could significantly inhibit NA(H7N9-WT). NA(H7N9-R292K) showed high-level resistance to both drugs (34-fold and 625-fold) and NA(H7N9-H274Y) was sensitive to both (1.3-fold and 9-fold). These results indicated that both oseltamivir and zanamivir could be used for patients infected with the H7N9 virus. However, when patients carried the H7N9 virus with a NA R292K mutation, other medications would be preferred over oseltamivir or zanamivir.
Antiviral Agents ; pharmacology ; Humans ; Influenza A Virus, H7N9 Subtype ; drug effects ; enzymology ; genetics ; Influenza, Human ; virology ; Microbial Sensitivity Tests ; Mutation ; Neuraminidase ; antagonists & inhibitors ; genetics ; metabolism ; Oseltamivir ; pharmacology ; Viral Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Zanamivir ; pharmacology

Composition of the polysaccharide from Bupleurum scorzonerifolium Willd. was studiedby high resolution capillary gas chromatography after hydrolysis and followed by acetylation and esterifica-tion of the hydrolysate. The results showed that it is composed of arabinose, ribose, D-xylose, D-man-nose, D-glucose and D-galactose, with a molar ratio of 0. 106 : 2. 652 : 4. 070: 0. 519 : 0. 930 : 2. 518.

Objective To investigate the expression of macrophage migration inhibitory factor (MIF) and CD_(74), the receptor of MIF, in preeclamptic placenta and its correlation with the pathogenesis of preeclampsia.Methods From March 2008 to November 2008,69 preeclamptic women who delivered in the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College,were recruited,including 33 women with mild preeclampsia (MPE group) and 36 women with severe preeclampsia (SPE group).Another 43 healthy pregnant women were taken as control group.Immunoturbidimetry was applied to measure the concentrations of C-reactive protein (CRP) in maternal blood.The expressions of MIF and CD_(74) in placenta were tested with immunohistochemistry and the expressions of MIF mRNA and CD_(74) mRNA were detected by semiquantitative RT-PCR.The relationship between maternal blood level of CRP and MIF mRNA and CD_(74) mRNA in placenta was analyzed in the MPE and SPE group.Results (1) MIF and CD_(74) were expressed in the placenta of all pregnant women in the 3 groups, as shown in brown-yellow color, and significantly higher expression was found in the MPE and SPE group.(2) The expression of MIF mRNA and CD_(74) mRNA in the MPE group (0.70±0.13 and 0.96±0.16), SPE group (0.88 ± 0.12 and 1.08 ± 0.15) were significantly higher than in the control group (0.67 ± 0.11 and 0.83 ± 0.14) (P < 0.01), and statistical significance was also found between the MPE and SPE group (P <0.01).(3)The maternal blood concentrations of CRP in the MPE and SPE group were significantly higher than in the control group [(15.3±7.0) mg/L and (21.6±9.1)mg/L vs (4.8 ± 1.8) mg/L, P <0.01] , and significant difference was also found between the MPE and SPE group (P <0.01).(4) In the two preeclamptic groups, the blood concentrations of CRP were positively correlated with the expression of both MIF mRNA(r =0.67 ,P <0.01)and CD_(74) mRNA(r =0.83 ,P <0.01) in placenta.Positive correlation was also found between the levels of MIF mRNA and CD_(74) mRNA in placenta (r =0.93 ,P < 0.01).Conclusions Overexpression of MIF and CD_(74) in the placenta may up-regulate the CRP level in maternal blood, resulting in systemic inflammatory reaction and vascular endothelium damage which may be involved in the pathogenesis of preeclampsia.

This study is to establish a cell-based model targeting to neuraminidase (NA) of the 2009 H1N1 influenza A virus. NA is an influenza virus structural protein with enzymatic activity of the cleavage of HA-sialic acid interaction to release new viral particles from cells. A model of HIV-1 (pNL4-3.Luc.R(-)E(-)) based pseudovirions packed with HA [hemagglutinin, A/VietNam/1203/2004 (H5N1)] and NA [A/California/04/2009 (H1N1)] was established to evaluate compounds activities on NA function. The viral release can be blocked by neuraminidase inhibitors, oseltamivir and oseltamivir carboxylate, with IC50 of (61 +/- 31) nmol L(-1) and (5.5 +/- 2.9) nmol L(-1) respectively. A point mutation of H275Y on NA leads oseltamivir-resistance. This corresponding mutation was introduced into the system which was also confirmed by oseltamivir and oseltamivir carboxylate.

Actins ; metabolism ; Angiomyolipoma ; metabolism ; pathology ; surgery ; Epiglottis ; Factor VIII ; metabolism ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged

Objective To study the mechanism of miR-548a-3p targeting MMP-2 inhibits the metastasis and invasion of gastric cancer cells.Methods Bioinformatics were used to search those miRNAs targeting MMP-2.The level of miR-548a-3p in gastric cancer tissues were detected by qRT-PCR and the expression of MMP-2 were detected by Western blot.The level of miR-548a-3p in three reconstructed gastric cancer cell lines were detected by qRT-PCR and the expression of MMP-2 were detected by Western blot.miR-548a-3p targeting the 3'-UTR of MMP-2 were detected by Luciferase reporter assay detected.And the changes of ability of metastasis and invasion were examined by Transwell experiment before and after transfection.Results There were high expression of miR217 and low expression of MMP-2 in human poorly differentiated gastric cancer cell line BGC-823.There were low expression of miR-548a-3p and high expression of MMP-2 in human middle and high differentiated gastric cancer cell line MKN-28 and SGC-7901 (P ＜ 0.05).The result of Luciferase reporter assay showed that miR-548a-3p can targeting the 3’-UTR of MMP-2.There were high expression of miR-548a-3p and low expression of MMP-2 in MKN28-miR-548a-3p mimics.There were low expression of miR-548a-3p and high expression of MMP-2 in others reconstructed three cell lines.There were smaller invaded cells in SGC-7901-miR-217 mimics than in others three cell lines (P ＜ 0.05).Conclusions miR-548a-3p can target the 3'-UTR of MMP-2 and down regulate its expression and inhibit the ability of invasion of gastric cancer cells.

Methionine synthase (MS, EC2.1.1.13), a key enzyme in the folate metabolism area catalyzing methyl transfer from N5-methyltetrahydrofolate to homocysteine to give tetrahydrofolate and methionine, takes a core position in folate cycle, one-carbon-unit transfer and sculpture amino acid pathways. Cobalamin-dependent methionine synthase was purified from rat liver. The enzyme was purified 609-fold to near homogeneity by batch chromatography on DE-52, anion-exchange chromatography on Q Sepharose Fast Flow and CHT-I hydroxyapatite column and was identified by SDS-PAGE and Western blotting. The enzyme activity was determined by spectrophotometric assay. In addition, the influencing factor and optimal reaction condition were performed. The steady state kinetic of rat liver methionine synthase was similar to that of other mammalian cobalamin-dependent methionine synthase which employed a Ping-Pong mechanism. The result indicated that cobalamin-dependent methionine synthase purified from rat liver is suitable for screening and studying methionine synthase specific inhibitors.