Objective To assess roles of standardized patients (SPs) in training of communication skills for resident general practitioners (GP). Methods Fifty-two resident GPs were trained for communication skills in consultation with SPs during May to July, 2008, including information feedback from SPs, lectures and group discussion coordinated by instructors on problems developed during residents' clinical encounters. Communication skills of the trainees with patients was assessed before and after training, respectively, including building physician-patient relationship, collecting information and providing feedback, understanding complaints from patients' perspective, sharing information and obtaining informed consents and closing consultation. Assessment was scored and proportion of trainees qualified for consultation with SPs were estimated. Results Total score of the trainees was higher after training than that before it (76 ± 13 vs. 49 ± 12, P ＜ 0. 01 ), and 86. 5 percent of them qualified for their communication skills after training, as compared to that of 15.4 percent before it. Proportions of the trainees qualified for building physician-patient relationship, collecting information and providing feedback, understanding complaints from patients' perspective, sharing information and obtaining informed consents during consultation were higher after training than those before it (90. 4% vs. 59. 6%, 65.4% vs. 5.8%, 38.5% vs. 11.5%, 86. 5% vs.17.3% and 65. 4% vs. 3. 9%, respectively, all P ＜0. 01 ). Scores of physician-patient communication in the trainees correlated with those of their consultation skills both before and after training ( r = 0. 55 and r =0. 63, both P ＜ 0. 01 ). Increase in scores of physician-patient communication and consultation skills in trainees after training also associated with that in those of clinical skills ( r = 0. 62, P ＜ 0. 01 ). Conclusions Application of SPs can effectively enhance communication skill of resident general practitioners, thus improve their competencies in clinical consultation.

Objective To assess standardized patients (SP) in resident general practitioners training for their consultation skills to explore its role in clinical competence training. Methods SP are employed in standardized training for 52 resident general practitioners in Shanghai during May to July 2008 to assess trainees' clinical consultation skills, including medical history taking and physical examinations, as well as clinical competencies of trainers including medical history taking, physical examinations, clinical thinking (diagnosis, differential diagnosis and treatment plans), health education, medical record writing,and reception time management. Feedback and appraisal for their consultation skills were provided by SP later. Problems encountered in trainees' consultation were discussed in group manner with their trainers.And two mouths later, assessment with SPs was undergone again for their consultation skills to compare their scores in consultation skills and percentages of them passing the assessment before and after training. Results Trainees were enrolled in the training aged (28±1 ) years in average ( with a ratio of men to women of 19/33 ). Percentage of trainees passing the assessment by SP for consultation skills was significantly higher after training (88.5%) than that before it (46. 2% ) ( P < 0. 01 ). Percentage of trainees with scores passing assessment by trainers was also significantly higher after training than that before it (86.5% vs. 51.9%, P < 0.01 ), with a percentage of those qualified for clinical skills and health education after training of 80. 8% and 88.5%, respectively, as compared to those of 46. 2% and 67. 3%,respectively before it, ( P < 0. 01 ), and with a percentage of trainees qualified for clinical thinking, out-patient medical record writing and reception time management not changed after training (71.2%, 100. 0%,100. 0%, respectively) as compared to those before it (51.9%, 100. 0% and 100. 0%, respectively)(P > 0. 05 ). Conclusions Application of SP can effectively enhance consultation skills of the trainees,especially in their medical history taking and physical examinations. However, there is still rooms to improvein their clinical thinking.

Baculovirus DNA polymerase gene belongs to an early gene of baculovirus. It is a necessary gene required for replication of virus in insect cells. It can encode DNA polymerase induced by virus. In the process of replication, DNA polymerase can bind to homologous regions and non-homologous regions, which are believed to act as the origins of virus DNA replication with other replication factors. In addition, DNA polymerase has advantages over occlusion protein and egt gene for resolving deep branching taxonomic relationships of baculovirus phylogenies.

With IL-6R as target, a new compound 2460A was identified from fungus using HTS screening model. The taxonomics of the produced strain was confirmed to be Trichoderma hazianum rifai after sequencing analysis of rDNA-ITS (internal transcribed spacer). Results showed that this compound has a binding activity on IL-6R competed with IL-6, thus it is a new ligand of IL-6R originating from microbe. With MTT assay, the anti-tumor activities of 2460A were demonstrated on CM126 and HT-29 cell lines separately, the IC50 are 2.17 x 10(-5) mol x L(-1) and 1.8 x 10(-5) mol x L(-1) respectively. The compound affected lightly the HT-29 cell cycle at S phase. Studies for the anti-tumor activity of 2460A in vivo are in progress in our lab.

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection

To explore the anti-tumor proliferation activity of human interleukin-29 (hIL-29) variant and based on bioinformatics analyzed data of hIL-29, a mutant gene hIL-29(mut33,35) was amplified by site-directed mutagenesis and megaprimer PCR. The hIL-29(mut33,35) was inserted into an eukaryotic expression plasmid pPIC9K and successfully expressed in Pichia pastoris GS115. A recombinant variant protein (rhIL-29(mut33,35)) was purified from the ferment supernatant of the engineering GS115. To observe the antineoplastic activity of the variant rhIL-29(mut33,35), a CCK-8 reagent was used to detect the anti-proliferation effect. Results show that it has strong anti-proliferation effect when acted on liver cancer cell BEL7402, colon cancer cell HCT8 and gastric cancer cell SGC7901. The inhibition ratios of the three tumor cells were (30.99 ± 1.58)%, (22.47 ± 1.37)% and (32.05 ± 2.02)%, respectively. In high dose group, the anti-proliferation effect of the rhIL-29(mut33,35) was stronger than that of wild type rhIL-29 (P < 0.01). This indicates the variant rhIL-29(mut33,35) has potential development value for medicine.
Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; drug effects ; Humans ; Interleukins ; biosynthesis ; pharmacology ; Liver Neoplasms ; pathology ; Mutagenesis, Site-Directed ; Pichia ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; pharmacology

Objective To investigate the role of opioid receptors in the protective effects of isoflurane-induced delayed preconditioning against myocardial ischemia-reperfusion (I/R) injury in rabbits. Methods Forty male New Zealand white rabbits weighing 2.0-2.5 kg were randomly assigned into 4 groups ( n = 10 each) : group I sham operation (S); group II I/R; group Ⅲ isoflurane + I/R (Iso) and group IV Iso + naloxone + I/R (Nal). Myocardial I/R was induced by 40 min occlusion of left anterior descending branch (LAD) of coronary artery followed by 120 min reperfusion. In group Ⅲ (Iso) 2% isoflurane in 100% O2 was inhaled for 2 h and I/R was produced 24 h later. In group IV (Nal) naloxone 6 mg/kg was given iv 10 min before 2 h of 2% isoflurane inhalation and I/R was produced 24 h later. At the end of 120 min reperfusion, infarct size (IS) and area at risk (AAR) were determined by Evan's blue and TTC staining. Myocardial ultrastructure was examined by electron microscopy. The phosphorylated p38MAPK protein expression in myocardium was determined by Western blot. Results The IS was significantly smaller in group Iso ( Ⅲ ) ( 19.7% ± 2.8%) than in I/R group ( II ) (37.8% ±1.7%) (P<0.05). The phosphorylated p38MAPK protein expression in myocardium was significantly lower in group Iso than in group I/R. Microscopic examination showed less myocardial damage in Iso group than in group I/R. The protective effects of delayed preconditioning by isoflurane was prevented by naloxone pretreatment. ConclusionOpioid receptors may be involved in the protective effects of delayed preconditioning by isoflurane against myocardial I/R injury.

Aim To observe the effect of hemodialysis on pharmacokinetics of sparfloxacin in thepatients contracting chronic renal failure. Methods Sparfloxacin concentrations inserum and urine of hemodialysis and non-dialysis patients were measured with a highperformance liquid chromatography method after administration a single oral dose of 200mg sparfloxacin. The pharmacokinetic parameters were computed with the programPKBP-N1.Results The main pharmacokinetic parameters in hemodialysis group wereT1/2(ka) - (1. 25 ?0. 57) h, T1/2(?) = (11. 88?4. 13) h, Tpeak = (4. 18 ? 0. 78) h,Cmax = (0.80 ? 0. 17) mg? L-1 and AUC0-= (6. 90 ? 3. 25) mg?h?L-1, while innon-dialysis group were T1/ 2(ka) = (1. 12 ? 0. 42) h, T1/ 2(?) = (15. 93 ? 5. 20) h, Tpeak =(3. 88 ? 0. 75) h, Cmax = (0. 69 ? 0. 37) mg?L -1, AUC0-= (10.05 ? 4. 13) mg?h?L-l. The original sparfloxacin discharge rats in urine within 24 h were (8. 98 ? 3. 92) % and(10. 58 ? 5. 64) % separately. T1/2(?) and AUC in hemodialysis group were markedly lowerthan in non-dialysis group (P

Background Statistic data revealed that different retinopathy of pre-term infants have different susceptibility to retinopathy of prematurity (ROP),which may be associated with polymorphism of vascular endothelial growth factor(VEGF) gene.Objective This study was to determine the association of polymorphisms of VEGF gene with the risk for ROP.Methods This research was approved by Ethics Committee of Hunan Children's Hospital,and written informed consent was obtained from the parents of patients.A prospective case-controlled study was designed.Ninety-nine ROP patients in Hunan Children' s Hospital and 88 pre-termed children without ROP were included from January,2006 to December,2009.Thirty-nine patients who received retinal photocoagulation or cryotherapy were included as the treatment group,and 60 untreated but spontaneously regressed ROP patients as the non-treatment group.No significant differences were seen in demography between with the ROP group and the without ROP group,or between the treatment group and the non-treatment group (all P＞0.05).2 mL of peripheral blood was collected for the extraction of DNA.Gene polymorphisms of VEGF-A+405 and VEGF-A936 were detected using the pyrosequencing method.Results No significant difference was found in the frequencies of the VEGF-A+405 gene polymorphisms between the ROP group and without ROP group (P =0.071,OR =0.675,95 ％ CI =0.444-1.026).Also no significant difference was found in the frequencies of the VEGF-A936 gene polymorphisms between with the ROP group and without the ROP group (P =0.118,OR =0.768,95 ％ CI=2.823-4.614).However,the frequencies of the VEGF-A+405 gene polymorphisms were significantly higher in the ROP treatment group than the non-treatment group (P＜0.01,OR--0.857,95 ％ CI =5.239-14.024),and VEGF-A936 gene polymorphisms was also significantly higher in the ROP treatment group than the non-treatment group (P =0.000,OR =3.609,95 ％ CI =0.711-0.829).Conclusions There is no association between the VEGF-A+405/VEGF-A936 single nucleotide polymorphism with the risk of ROP,but polymorphisms of VEGF gene may be correlated with the prognosis of ROP.The carrier of VEGF-A +405 /VEGF-A936 allele is more susceptible to ROP progression.

Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.