OBJECTIVE To investigate the affinity of 20 Klebsiella pneumoniae(KPN)strains.METHODS Twenty nine genes were detected from 20 KPN strains.They were sixteen kinds of beta-lactamase genes,six kinds of aminoglycosides modification enzyme genes,chlorhexidine-sulphanilamide drug resistance genes,TMP drug resistance genes;three kinds of integrase genes and Tn21/Tn501 transposon genetic mark were detected.And they were used as molecule marker for the sample clustering analysis and assay of the detected results.RESULTS The beta-lactamase genes,aminoglycosides modification enzyme genes,chlorhexidine-sulphanilamide drug resistance gene,TMP drug resistance gene,integrase genes and Tn21/Tn501 transposon genetic mark were all detectable.Clone transmission was shown through the sample clustering analysis.CONCLUSIONS The resistance to the beta-lactam and aminoglycoside antibiotics of the 20 KPN strains is intimately correlated to the generation of beta-lactamases and amino glycosides modification enzymes,which could be clone transmitted.

Objective To study role of color Doppler ultrasonography in predicting the benignity and malignancy of the peripheral hypoechoic lesion of the prostate.Methods Seventy-seven patients who had peripheral hypoechoic lesions were detected with transrectal color Doppler ultrasonography.The black and white color ratio (BCR) in peripheral hypoechoic was calculated by color histogram and compared with prostate BCR.The amount of flow signal was considered increasing if BCR in the lesions was higher than 5% in their prostate.Results Transrectal ultrasound-guilded biopsy of the hypoechoic lesions revealed prostate cancer in 51 patients and benign prostate hypertrophy in 26 patients.The increase of flow signal was in 50 patients,41 of them were prostate cancer.For an increased flow signal within a peripheral hypoechoic lesion as a signal of prostate cancer,color Doppler ultrasonography has a sensitivity of 80.4% ,a specificity of 65.4% ,and a positive predictive value of 82.0% .Conclusions Color Doppler ultrasonography through rectum on blood flow in peri-prostate hypoechoic nodules with BCR account for their rich degree in malignancy and benignity differentiation.

Two systems which covered nearly 1. 1 billion people in China’s social health insurance, namely the New Rural Cooperative Medical Scheme ( NRCMS ) and the Urban Residents’ Health Insurance System ( URHIS ) , have been depended mainly on government support with high percentage of premium subsidies. This paper deals with the current situation and probes into the main subsidy policy problems by making an overall analysis on the policies of 31 cross-country provinces , and in result points out four problems including the long-term mechanism shortage, in-distinct responsibility between governments at different levels, the equity deficiency and the subsidies inefficiency. Finally, the paper concludes with enlightening some corresponding policies and suggestions.

Objective To assess the role of hip ultrasonography in the early screening and following up of developmental dysplasia of the hip (DDH). Methods A total of 1324 hips of 662 infants younger than 6 months underwent ultrasonography with Graf method. The sonographic appearances of bony roof, superior bony rim, cartilaginous roof, as well as α and β angle measurements were classified according to the Graf method. Risk factors such as gender, sides, fetal position, and swaddling used were assessed. Following up were performed with ultrasonography in the cases of immaturity, dysplasia and dislocation that needed to be monitored or treated. Results Detection rate of selective ultrasound in screening of DDH of the hip was 7.42%. Both gender and fetal position was the risk factor of DDH. Twenty-seven infants with dysplasia hip and five with dislocation hip were followed-up with ultrasonography during treatment process. Thirteen infants with dysplasia turned to normal after abduction exercises, while other fourteen received treatment in Pavlik harness for no improvement in ultrasonography. All infants of dislocation received operation eventually because of failing to Pavlik harness. Conclusion Ultrasonography can objectively assess the development of DDH, monitor the course of following-up and treatment of DDH, and serve as a main tool in the early screening and following up of DDH in infants.

Objective; To investigate the immunolocalization and significance of Notch-2 expression in the process of dental pulp repair after injury. Methods: An experimental animal model of injury-induced pulpitis was established to observe the time-sequenced alteration of the expression of Notch-2. Results: Three days post-operation, weak positive staining of Notch-2 was observed in pulp mesenchyme cells and pulp fibroblasts but not in vascular endothelial cells or odontoblasts. Five days post-operation, strong Notch-2 reactivity was found in subodontoblasts as well as newly bom capillary endothelial cells. Seven days after cavity preparation, Notch-2 staining became weaker in pulp mesenchyme cells and capillary endothelial cells, but stronger positive staining was found in odontoblasts. Two weeks post-operation, weak Notch-2 staining was seen in pulp mesenchyme cells and subodontoblastic layer cells and was absent from odontoblasts. Notch-2 immunoreactivity was completely absent in intact rat dental pulp. Conclusion: Notch-2 is strikingly up-regulated in dental pulp mesenchyme cells in the early days after dental injury and then shows progressively up-regulation in odontoblasts. Properly regulated activation of Notch signaling pathway is important for controlling cell fate and maintaining the correct balance among cell proliferation, differentiation and apoptosis during dental pulp repair process.

Objective To evaluate the efficacy of emergency endoscopic intervention in acute obstructive suppurative cholangitis (AOSC) complicated with septic shock.Methods A total of 54 patients with AOSC and septic shock who underwent therapeutic emergency ERCP were included in this retrospective study,and were evaluated by the shock index (SI).Results ERCP was performed for all patients in 24hours after hospitalization,and the average ERCP operation time was 23.8 ± 12.5 min.All 54 patients underwent EST,46 of whom received ENBD,7 biliary stenting and 1 transferred to surgery due to bleeding.The post-ERCP mortality rate was 0,and the complications included 1 case of pancreatitis and 2 cases of pneumonia.The positive rate ofGram-Negative bacillus before ERCP was 46.9％ (15/32).The SI before ERCP was 1.250 ±0.200,which decreased to 0.950 ±0.119 at 2hr after the procedure (P ＜0.001 ),and decreased further to 0.598 ± 0.099 ( P ＜ 0.001 ) at 24 hours after ERCP.Conclusion Therapeutic emergency ERCP is of great importance in the treatments for AOSC complicated with septic shock.

AIM:To synthesize a safe , efficient and targeted nanoparticulate carrier for siRNA delivery to pan-creatic cancer cells .METHODS: Iron oxide nanocrystal with carboxylic acid group-polyethyleneimine ( IONP-PEI ) was synthesized and investigated as a nonviral carrier of siRNA to the pancreatic cells .The size, surface and charge using zeta potential were characterized .The perfect charge ratio between amino groups of IONP-PEI and phosphate groups of siRNA ( N/P) was determined by the transfection efficiency detection , gel retardation assay and MTS assay .An antibody-directed nonviral vector , scFvCD44v6-IONP-PEI nanoparticle attaching to the cancer-associated CD44v6 single-chain variable frag-ment, was constructed as a cancer-targeting nanocarrier for siRNA delivery .Prussian blue staining and immunofluorescent staining were performed to detect the distribution of scFv CD44v6-IONP-PEI/siRNA complexes in the cells .The transfection efficiency , fluorescence intensity and the expression of KRAS at mRNA and protein levels in the cells transfected by IONP -PEI/siRNA and scFv CD44v6-IONP-PEI/siRNA were detected by flow cytometry , fluorescence microscopy , real-time PCR and Western blotting, respectively.RESULTS:The mass ratio of IONP to PEI was 0.75.The suitable ratio of N/P was 20. The averaged size and surface zeta potential of IONP-PEI/siRNA in deionized water were (51.3 ±2.2)nm (diameter) and (21.73 ±8.07)mV, respectively.Red fluorescence was seen in both targeting and nontargeting groups , which clearly re-vealed the intracellular distribution of siRNA and delivery agents .Transfection efficiencies in targeting and nontargeting groups were (89.75 ±1.81)%and (59.87 ±4.52)%, respectively.Down-regulation of the KRAS mRNA in Panc-1 cells transfected with siKRAS by scFvCD44v6-IONP-PEI and IONP-PEI was up to (34.02 ±6.15)%and (51.09 ±6.70)%, re-spectively .The protein level of KRAS was lower in targeting group than that in nontargeting group .CONCLUSION:scFvCD44v6-IONP-PEI is a safe and efficient nanoparticulate carrier for gene delivery .It is more effective to transfer siRNA into the cells and mediate gene silencing effect in vitro than the nontargeting group .

Objective To establish a polymerase spiral reaction (PSR) method for rapid detection of influenza A/H1N1 virus.Methods Six sets of primers were designed for influenza A/H1N1 virus HA gene, and the results were determined with real time kinetic turbidimetric assay and colorimetry method.Results and Conclusion The best primers were selected from six sets of primers, and the best temperature was determined as 65 degrees Celsius.Further experiments showed that the best primer had good specificity for detection of influenza A/H1N1 virus,without cross reactions with 14 other respiratory tract pathogenic nucleic acids.The sensitivity was up to 100 copies,and consistent with that of PCR.So a PSR method is established for rapid detection of the influenza A/H1N1 virus, which is simple, quick, highly specific and sensitive,and especially applicable to field and grass-roots units.

Objective To investigate the neuroprotective effect and its mechanism of p38 mitogenactivated protein kinase inhibitor after subarachnoid hemorrhage (SAH).Methods Twenty-seven SPF-grade adult male SD rats were selected to induce a SAH model using the prechiasmal pool blood injection.Three dead rats were excluded.Twenty-four rats were randomly divided into four groups:sham operation group,SAH group,dimethyl sulfoxide (DMSO) group,and DMSO +p38 inhibitor group (n =6 in each group).Western blot was used to detect the expression levels of p38,phosphorylation p38,Parkinson's disease protein 7 (DJ-1),autophagy associated gene 5 (Atg5),autophagy adaptor protein p62,microtubule-associated protein Ⅰ Light Chain 3 (LC3-Ⅰ),microtubule-associated protein Ⅱ light chain 3 Ⅱ (LC3-Ⅱ),and the Garcia neurological function score was used to judge the nerve injury.PC12 cell oxygenated hemoglobin was used to induce an in vitro SAH model.They were completely randomly divided into four groups:sham operative group,SAH group,DMSO group,and DMSO + p38 inhibitor group.Fluorescent probe JC-1 was used to observe the changes of mitochondrial membrane potential.Results (1) There were significant differences in the expression of p38,phosphorylation-p38 and DJ-1 in rat brain tissue among the 4 groups (F values were 94.959,150.293 and 698.476,respectively,all P ＜ 0.01).There were significant differences in mitochondrial membrane potential in PC12 cells among the 4 groups (F value was 24.989,P ＜ 0.01).There were significant differences in the expression levels of autophagy related protein LC3-Ⅱ/LC3-Ⅰ ratio,Atg5 and p62 in rat brain tissue among the 4 groups (F values were 235.319,110.490 and 36.311,respectively,all P ＜ 0.01).There was significant difference in nerve function score among the 4 groups (F value was 25.550,P ＜ 0.01).(2) Compared with the sham operative group,the expression levels of p38,phosphorylation-p38 and DJ-1 were upregulated significantly after SAH (from 0.43 ±0.06,0.41 ±0.02 and 0.07 ±0.01 to 0.61 ± 0.08,0.79 ± 0.07 and 0.17 ± 0.03,respectively,all P ＜ 0.01),mitochondria membrane potential depolarization (from 8.29 ±0.28 to 9.23 ±0.42,P ＜0.01);upregulation of Atg5 expression and increase of LC3-Ⅱ/LC3-Ⅰratio (from 0.23 ± 0.04 and 0.25 ± 0.04 to 0.47 ± 0.04 and 0.46 ± 0.04,respectively,all P ＜ 0.01),down regulation of p62 expression (from 1.09 ± 0.14 to 0.54 ± 0.10,P ＜ 0.01);neurological score was decreased (from 17.5 ± 0.6 to 11.3 ± 2.7,P ＜ 0.01);p38 inhibitor was significantly down regulated the expression of phosphorylation-p38 after SAH (from 0.79 ± 0.07 to 0.47 ± 0.04,P ＜ 0.01),the expression of DJ-1 was up-regulated (from 0.17 ± 0.03 to 1.02 ± 0.06,P ＜ 0.01),mitochondrial membrane potential recovery (from 9.23 ±0.42 to 8.47 ±0.36,P ＜0.01),cell autophagy related protein LC3-Ⅱ/LC3-Ⅰ ratio and Atg5 were upregulated(from 0.46 ±0.04 and 0.47 ±0.04 to 0.77 ± 0.06 and 0.95 ± 0.12,all P ＜ 0.01),p62 expression returned to the levels of SAH group (from 0.57 ± 0.09,to 0.54 ± 0.10,P =0.650),and the neurological score was significantly improved (from 11.3 ± 2.7 to 15.5 ± 1.0,P ＜0.01).Conclusions After SAH,the p38 inhibitor downregulates the activity of2 phosphorylation p38.It may inhibit abnormal autophagy and maintain mitochondrial function by up-regulating the expression of DJ-1 protein,and then play a neuroprotective function.

Objective:To study on the effects of gastrointestinal function and portal vein hemodynamics applicated with abdominal hot compressing with evodiae fructus and crude salt after hepatectomy.Methods:A total of 60 patients who underwent hepatectomy were randomly divided into 2 groups by random number table method, with 30 in each group. The control group was treated with conventional basic western medicine, while the treatment group was treated with medicinal evodiae fructus and crude salt hot compress on abdomen on the basis of the control group. The portal vein diameter (PVD), portal venous flow velocity (PVV), recovery time of main clinical indexes, clinical symptom scores and liver function indexes were dynamically monitored at different observation time points.Results:The scores of abdominal distension, nausea and vomiting in the treatment group were significantly lower than those in the control group 3 days after treatment ( t values were -3.489 and -2.740, respectively, all Ps<0.05). The recovery time of bowel sounds, first exhaust time and first defecation time in the treatment group were significantly earlier than those in the control group ( t values were -3.622, -4.297 and -4.151, respectively, all Ps<0.01). With the extension of treatment time, ALT in 2 groups showed a gradual downward trend ( P<0.05 or P<0.01), DBIL in control group was significantly higher at 3 days after treatment than before ( t=-2.157, P=0.039), and TBIL was significantly lower at 7 days after treatment than before ( t=2.175, P=0.038). The PVD ( t values were 3.528, 2.160) and PVV ( t values were 11.096, 4.264) of the treatment group were significantly higher than those of the control group 3 and 5 days after treatment ( P<0.01 or P<0.05). Conclusion:Early application of abdominal hot compressing with evodiae fructus and crude salt hot compress on abdomen after hepatectomy can improve the portal vein blood circulation and promote the rehabilitation of gastrointestinal function in patients with hepatectomy.