Problem-based and student-centered learning patten brings new challenges to the both sides of teaching and learning.Clinical probation is the first step of medical students.The problem-based learning patten is used to clinical probation and achieves good results.It helps to improve medical students' quality and should be applied in China medical education.

Objective To establish a dot blot hybridization technique for rapid detection of staphylococci and methicillin-resistant staphylococci.Methods Three pairs of primers were designed according to nuc gene of staphylococcus aureus,mecA gene of methicillin-resistance,tuf gene of staphylococci.Specific DNA probes were synthesized by polymerase chain reaction and labeled with biotin.The bacterial DNA inoculated on nitrocellulose filter was hybridized with these probes.The sensitivity and specificity were detected.Results The DNA probes with 270bp,310bp and 370bp were amplified by the three pairs of primers respectively.The probes were specific.Among 50 clinical isolates of staphylococcus aureus tuf and nuc gene were all positive and mecA gene in 22 isolates were positive.Positive rate of tuf,nuc and mecA gene in 30 staphylococcus epidermidis were 100%,0 and 30% (9/30) respectively.No hybridization in other non-staphylococci occurred.The established method could detect as low as 1ng of bacterial DNA.Conclusion The dot blot hybridization is of high value in rapid,effective identification of methicillin-resistant staphylococci.

Objective:To explore influence of monotherapy or combined use of telmisartan and pyridoxamine on aor-tic remodeling in spontaneously hypertensive rats (SHR)and its possible mechanism.Methods:A total of 48 male SHE were randomly and equally divided into hypertension control group,telmisartan group,group,and telmisartan+ group (combined treatment group). Kyoto Wistar rats of the same age and gender were regarded as normal blood pressure control group (normal control group). Thoracic aortic section were examined by related staining af-ter 16 weeks intervention to calculate the ratio of aortic wall thickness to radius of lumen (Tw/Rl),the ratio of wall area to lumen area (W/L),and the area ratio of media elastic fiber/collagen fiber. Concentrations of related en-zymes and receptor etc. of abdominal aortic were measured.Results:Compared with hypertension control group, there was significant rise in ratio of media elastic fiber/collagen fiber area and significant reduction in media collagen fiber/media area ratio in telmisartan group,pyridoxamine monotherapy group and combined treatment group,and there were significant decrease in Tw/Rl [(0.17±0.02)vs. (0.12±0.01)]and W/L [(0.29±0.03)vs. (0.22± 0.02)]ratios in combined treatment group,P <0.05 or <0.01;immunohistochemistry indicated that there were significant reductions in thoracic aortic receptor of advanced glycation end products (RAGE) [(0.24±0.03)vs.(0.17±0.03)]and extracellular signal-regulated kinase 1/2 (p-ERK1/2 )expression [(0.63 ± 0.06)vs. (0.37± 0.04)]in combined treatment group,P <0.05,<0.01. Fluorescence quantitative PCR indicated that medication can significantly reduce nicotinamide adenine dinucleotide phosphate (NADPH)oxidase subunit p47phox mRNA ex-pression (P <0.01 all),especially in combined treatment group (P =0.001).Conclusion:Combined use of telmis-artan and pyridoxamine is superior to the single use of either drug on improving thoracic aortic remodeling in SHR, the mechanism may be related to it reduces local expression of RAGE and p-ERK1/2 ,and inhibits oxidase subunit p47 of NADPH.

Objective To explore the ill experience of patients with deep vein thrombosis (DVT),and supply reference for making corresponding intervention.Methods The qualitative phenomenological study method was adopted in this study.Self-structured in-depth interviews were conducted with 12 patients with deep vein thrombosis and data were collected for analysis.Results After repeated refinement of the collected data,the early ill experience of deep vein thrombosis could be extracted as 4 topics.Conclusions According to the ill experience of patients with DVT,nurses can provide targeted intervention for such diseases to instruct clients to rehabilitate rapidly and improve the patients' quality of life.

Objective To investigate the effects and the mechanism of telmisartan and pyridoxamine on oxidative stress in spontaneously hypertensive rats(SHR).Methods SHRs(male,20 weeks of age) were randomly divided into four groups (n＝ 12 for each):hypertension control (HC) group (2 ml of distilled water),telmisartan group[T,6 mg/(kg · d)],pyridoxamine group[P,200 mg/(kg · d)]and combined group(TP,6 mg/kg telmisartan and 200 mg/kg pyridoxamine per day).Treatments were continued for 16 weeks.The normal control group included 13 WKY rats and received gastric lavage with distilled water.SBP of tail artery was measured during the intervention ervey 2 weeks.The levels of AGEs,SOD and MDA were measured by ELISA,xanthine oxidase and thiobarbituric acid methods after the intervention.Expressiones of NF-κBp65,ERK1 and ERK2 in renal tissue were detected by immunohistochemistry.Expression of RAGE in the renal cortex was investigated by Western blot.Results SOD activity was decreased in the HC group.The levels of AGEs,MDA,RAGE and the activations of NF-κBp65 and ERK1/2 were increased in the HC group (t＝4.53,5.52,2.93,al1 P＜0.05).After the 16 weeks' intervention,SOD activity was elevated in T,P and TP groups compared to that in HC group (P＜0.05).The positive expressiones of NF-κBp65,ERK1 and ERK2 were significantly reduced in T,P and TP groups compared to those in HC group (F＝20.13、148.82、18.70,all P＜0.05).All the positive expressiones of NF-κBp65,ERK 1and ERK2 were lowest in the TP group versus T and P groups (t ＝ 3.58、2.84,P ＜ 0.05).Conclusions Telmisartan and pyridoxamine can alleviate the oxidative stress in spontaneously hypertensive rats,which may result from the blocking effect of Ang Ⅱ,the reduction of AGEs-RAGE and inhibiting the signal pathways of ROS,NF-κBp65 and ROS-ERK1/2.

[ ABSTRACT] AIM:To explore the inhibitory effects of pantoprazole sodium on epithelial-mesenchymal transition and cisplatin resistance in lung cancer cells and the underlying mechanism .METHODS: Using MTT method, wound healing assay , Transwell experiment , Western blot , the differences of morphology , invasion ability , migration ability , drug sensitivity and protein expression between A 549/DDP cells and A549 cells were determined .The effect of pantoprazole so-dium on morphology , invasion ability , migration ability , drug sensitivity and protein expression in A 549/DDP cells were al-so observed.RESULTS: Compared with A549 cells, A549/DDP cells had higher invasion and migration abilities , and lower drug sensitivity , exhibited mesenchymal phenotype and activated c-Met/AKT/mTOR pathway .Pantoprazole sodium inhibited the abilities of invasion and migration , and reversed the mesenchymal phenotype , drug resistance and the c-Met/AKT/mTOR pathway activation in A549/DDP cells.Treatment with c-Met inhibitor SU11274, PI3K inhibitor LY294002 and mTOR inhibitor rapamycin had the same effects on A 549/DDP cells as that of pantoprazole sodium .CONCLUSION:Pantoprazole sodium inhibits invasion , migration, epithelial-mesenchymal transition and cisplatin resistance in lung cancer cells by down-regulating c-Met/AKT/mTOR pathways .

Objective To investigate the effects of the frozen embryo transplantation for patients with poor outcome of endometrial growth by using growth hormone (GH) intrauterine perfusion combined with replacement cycle in the treatment of thin endometrium.Methods This was a prospective study and study participants were consecutively recruited between Jun 2014 and September 2015.A total of 88 frozen thawed embryo transfer cycles was divided into two groups from the Reproductive Center of Hunan Provincial Maternal and Child Health Hospital.Group A were 63 hormone replacement therapy (HRT) cycles and Group B were 25 GH intrauterine perfusion combined HRT cycles.Results The endometrial thickness of 22 thin endometrium patients from Group B were increased above 7 mm on progesterone day.The endometrial thickness on transplant day of Group A was (9.28 ± 1.64) mm,which was significantly higher than Group B (7.9 ± 0.86) mm (P ＜ 0.05).The clinical pregnancy rate (50.79％ vs 52.0％),implantation rate 31.1％ vs 47.17％),miscarriage rate (9.38％ vs 15.38％) had no significant difference between Groups A and B.The endometrial thickness from 7 mm to 7.9 mm on transplant day,the clinical pregnancy rate (30.76％ vs 54.54％) had no significant difference in two groups (P ＞0.05),but the implantation rate of group A was significantly lower than that of group B (20％ vs 52.17％) (P ＜0.05).When the endometrial thickness was above 8 mm on transplant day,the clinical pregnancy rate (58.33％ vs 63.63％),implantation rate (36.36％ vs 50％) had no significant difference between groups A and B (P ＞ 0.05).Conclusions GH uterine cavity perfusion was a useful method for treatment of thin endometrium,and was helpful for improvement of endometrial thickness and receptivity,improved embryo implantation environment by assistance for HRT under the high estrogen levels.

Objective: To observe the effect of pyridoxamine and telmisartan on the indexes of myocardial remodeling in spontaneous hypertensive rats (SHR). Methods: A total of 48 male SHR at 22 weeks of age were randomly divided into 4 groups:①Hypertension (H) control group, the rats received distilled water 2 ml/d,②telmisartan (T) group, the rats received telmisartan 6 mg/(kg.d),③pyridoxamine (P) group, the rats received pyridoxamine 200 mg/(kg.d),④Combination (TP) group, the rats received both telmisartan and pyridoxamine. All animals were treated for 16 weeks and the systolic blood pressure (SBP) was measured before and after treatment. After treatment, the serum levels of nitric oxide (NO) and super oxide dismutase (SOD) were examined by chemiluminescent method, advanced glycation end-products (AGEs) was detected by ELISA, left ventricular weight index and collagen volume fraction (CVF) in myocardial tissue were calculated, the mRNA expression of myocardial receptor of advanced glycation end products (RAGE) was evaluated by real time RT-PCR. Results: Compared with H group, SBP levels were decreased in T and TP groups,P<0.01, while SBP in P group stayed similar,P>0.05. Compared with H group, the serum levels of NO and SOD were increased in T, P and TP groups,P<0.01; while compared with T group and P group, the levels of NO and SOD were further increased in TP group,P<0.05. Compared with H group, left ventricular weight index and CVF were decreased in T, P and TP groups,P<0.01; while compared with T group and P group, left ventricular weight index and CVF were further decreased in TP group,P<0.05. Pyridoxamine and telmisartan had the coordinative effect on left ventricular weight index and CVF, P<0.05. The serum levels of AGEs were lower in T group (5.99 ± 0.51) mg/L, P group (5.57 ± 0.91) mg/L and TP group (5.24 ± 0.63) mg/L than that in H group, (6.71 ± 0.50) mg/L,P<0.01; while compared with T group and P group, AGEs was further decreased in TP group,P<0.05. The mRNA expressions of RAGE were reduced in T group (0.035 ± 0.010), P group (0.036 ± 0.005) and TP group (0.024 ± 0.007) than that in H group (0.053 ± 0.010),P <0.01; while compared with T group and P group, the mRNA of RAGE was further reduced in TP group, P<0.05. Conclusion: Pyridoxamine could improve the oxidative stress and the indexes of myocardial remodeling independently from decreasing blood pressure in SHR. Combined intervention of P and T may break the coordination systems between AGEs-RAGE and renin-angiotensin, therefore further improve the indexes of myocardial remodeling in SHR.

Objective To analyze the correlation between emphysema extent measured by high resolution computed tomography (HRCT) and pulmonary function tests, symptom score in patients with chronic obstructive pulmonary disease ( COPD ) , and to study the value of HRCT in the emphysema quantification in the clinical evaluation of COPD patients. Methods 78 patients with stable COPD were recruited to take the HRCT scan , and emphysema extent was qualified by measuring the proportion of low attenuation area in the whole lung (LAA%). Correlations between LAA% and indices of pulmonary function test, bronchial dilation test, mMRC scale, CAT score and six minutes walking distance (6MWD) were assessed. Results LAA% was negatively correlated with FEV1/FVC and DLCO%pred, and the correlation coefficients were -0.759 and -0.589 (P <0.01), respectively. LAA% was positively related to mMRC score (r = 0.342, P < 0.01), and negatively asso-ciated with 6MWD (r = -0.365,P< 0.01). There was no association between LAA% and indices of bronchodila-tion test (⊿FVC, ⊿FVC%, ⊿FEV1, ⊿FEV1%) (P > 0.05). Conclusions The severity of emphysema measured by HRCT is well correlated with the clinical symptoms , pulmonary function tests and exercise capacity in COPD patients. It can be used to diagnose emphysema early and to evaluate the severity of the disease com-prehensively. Thus, the risk factors of COPD can be controlled and the prognosis of the patients can be im-proved.

Objective To evaluate the role of 131I SPECT/CT in post-surgical re-staging and recurrence risk stratification in patients with DTC and its impact on subsequent treatment strategy.Methods 131I-WBS and 131I SPECT/CT were performed at the same time 5 to 7 d after 131I treatment in 118 patients (33 males,85 females,average age 45 years) with DTC.Difference in the localization and qualitative diagnosis of 131I uptake lesions between 131I-WBS and 131I SPECT/CT were compared.Value of 131I SPECT/CT in the diagnosis of TNM staging,risk stratification and impact on the treatment strategy was evaluated.Paired χ2 test was used for data analysis.Results A total of 509 foci with 131I uptake were detected.131I-WBS found 449 foci with 131I uptake,354 of which (78.84％) were correctly diagnosed.131I SPECT/CT found 509 foci with 131I uptake,and 504(99.02％) were correctly diagnosed.The difference was statistically significant (χ2=21.51,P<0.01).131I-WBS changed the clinical staging in 13 cases with diagnostic accuracy of 5/13.131I SPECT/CT changed the clinical staging in 19 cases and with diagnostic accuracy of 19/19 (χ2=74.41,P<0.01).131I-WBS changed the risk stratification of 13 patients after operation and the accuracy was 5/13,the corresponding data were 22 and 100％(22/22) for 131I SPECT/CT (χ2=74.41,P<0.01).The treatment strategy was changed in 50 patients with 131I SPECT/CT.Conclusions Compared with 131I-WBS,131I SPECT/CT could provide more accurate positioning and qualitative information for 131I treatment and is more accurate in re-staging and risk stratification.