Objective To observe the effect of MG132 on the expression of extracellular regulated kinase 1/2 (ERK1/2) and connective tissue growth factor (CTGF) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.Methods RPMCs were isolated,cultured and passaged by trypsin,then identified.The second generation of cultured RPMCs were used in the experiment.RPMCs were divided into normal control group,high glucose (1.5％,2.5％,4.25％) for 24 hours,high glucose (2.5％) for 0,12,24,48 hours,incubated with MG132 (0.5,1,2 μmol/L) for half an hour and then with high glucose (2.5％) for 24 hours.ERK1/2 protein was detected by Western blotting,and CTGF protein in supernatant was detected by ELISA.Results Compared with the control group,the expression of p-ERK1/2 was significantly increased in the groups stimulated by high glucose (P ＜0.01),reached the peak at 24th hour (P ＜ 0.01),and then the expression decreased at 48th hour,but still was higher than that in the normal control group (P ＜ 0.01).CTGF protein expression of RPMCs induced by high glucose increased,in time-and dose-dependent manner (P ＜ 0.05).MG132 could significantly decrease the expression of ERK1/2 and CTGF induced by high glucose (P＜0.05).Conclusions MG132 can decrease the expression of p-ERK1/2 and CTGF in RPMCs induced by high glucose.The ubiquitin proteasome pathway participates in the development of peritoneal fibrosis,and blocking the way may contribute to the prevention of peritoneal fibrosis.

Objective To explore the folate receptor mediated (FRD) check and multi-function acetic acid white solution liquid based cervical cytology (TCT) application value in cervical cancer screening.Methods A total of 602 cases of patients was tested with FRD multi-function acetic acid white solution check,and TCT and cervical biopsy pathology examination.With the used of histopathological results as the gold standard,FRD multi-function acetic acid white analysis was compared with the TCT screening inspection results.Results For a total 602 patients with TCT screening,the positive rate was 21.8％ (131/602),including 36 cases of CIN Ⅰ level,41 cases of CIN Ⅱ level,24 cases of CIN Ⅲ level,and 30 cases of cervical invasive carcinoma.For the FRD multifunction white acetate solution screening,its positive rate was 23.8％ (143/602).No statistically significant difference was found between TCT and FRD screening (P ＞ 0.05).The missed diagnosis rate of FRD multi-function white acetate solution screening was 2.6％ in inflammation,and 21.1％ in cervical invasive cancer,and 3.8 ％ in CIN.The missed diagnosis rate of TCT screening was 7.2％ in inflammation,5.3％ in CIN Ⅰ,4.9％ in CIN Ⅱ,and 58.6％ in CIN Ⅲ]; whereas,its detection coincidence rate was 100％ in squamous cells carcinoma (SCC) and adenocarcinoma (AC).FRD multi-function acetic acid white solution screening had a sensitivity 80.92％,specificity 92.14％,positive predictive value 74.13％,and negative predictive value 95.59％.TCT examination had a sensitivity 90.84％,specificity 90.23％,positive predictive value 72.12％,and negative predictive value 97.25％.No significant difference was found between FRD and TCT methods (P ＞ 0.05).Conclusions FRD and TCT methods were both efficient in screening and evaluation for cervical lesions and cervical cancer.Because FRD method is limited in the deep tube for examination of cervical lesions; it cannot completely replace the TCT examination.However,FRD method is reliable,economic,and simple operation; it is suitable for primary hospitals census of cervical cancer

This paper introduces reformation of course system for five-year clinical medicine specialty in three aspects: orientation of course arrangement ,basic structure and major features of new teaching model. By adjusting the teaching plan and setting up new course system optimally, we have made an active attempt on new teaching model for bringing up high-quality medical talent.

Objective To observe the effect of silibinin on the expression of integrin linked kinase (ILK),transforming growth factor β1 (TGF-β1) and α-smooth muscle actin (α-SMA) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.Methods RPMCs were isolated,cultured and passaged by trypsin,then identified.The second generation of cultured RPMCs were used in the experiment.RPMCs were divided into normal control group,high glucose(1.5％,2.5％,4.25％)for 24 hours,high glucose (2.5％) for 12,24,48,72 hours,high glucose (2.5％) for 24 hours after silibinin (5,10,20 mg/L) preincubate for 2 hours.ILK and α-SMA mRNA were detected by real-time PCR.ILK protein was detected by Western blotting.TGF-β1 protein in supernatants was detected by ELISA.Results Compared with the control group,the expresssion of ILK,TGF-β1 and α-SAM was significantly increased in groups stimulated by high glucose (all P ＜ 0.05).Silibinin could significantly decrease the expression of ILK,TGF-β1 and α-SMA induced by high glucose (all P ＜ 0.05).Conclusions High glucose can up-regulate the expression of ILK,TGF-β1 and α-SMA.Silibinin can reverse these changes.

BACKGROUND:Specific down-regulation of interleukin 1 beta (IL-1β) may al eviate the pain behaviors effectively after peripheral nervous injury. Compared with smal interference RNA (siRNA), short hairpin RNA (shRNA) could inhibit the expression of target gene more stably and efficiently. However, simple shRNA could not enter target cel s to down-regulate target gene efficiently. Adenovirus vectors have wide host range, high infection efficiency and stable expression in host cel s.
OBJECTIVE:To construct recombinant adenovirus vector expressing shRNA targeting IL-1βgene and detect its effect on the expression of target gene.
METHODS:Three siRNAs were designed on the basis of the nucleotide sequence of IL-1βobtained from NCBI and then three shRNAs (shRNA1, shRNA2 and shRNA3) were synthesized. The annealed shRNA product and adenovirus vector pHBAd/U6/GFP digested by BamH I and EcoR I were connected to construct the recombinant adenovirus vector shuttle plasmid expressing shRNA targeting IL-1β. After sequencing, HEK 293 cel s were co-transfected by the shuttle plasmid and skeleton vector, and three recombinant adenovirus vector expressing shRNA targeting IL-1β(rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3) were packaged and amplified. Rats H9C2 cel s were infected by recombinant adenovirus vector expressing shRNA targeting IL-1βand fluorescence microscope was used to observe the infection efficiency. The effect of recombinant adenovirus vector expressing shRNA targeting IL-1βon the expression of target gene was detected by western blot assay.
RESULTS AND CONCLUSION:The sequencing results showed that the sequences of three shRNAs adenovirus vector shuttle plasmid were consistent with the sequences of three designed shRNAs. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 were constructed successful y. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 could down-regulate the expression of IL-1βin rat H9C2 cel s and the down-regulation effect of rAd/shRNA2 was the most significant.

Objective To establish blood purification system for rats and provide a safe and reliable experimental platform for further research of blood purification. Methods The right carotid artery and the contralateral jugular vein of adult male Sprague?Dawley rats were cannulated to creat vascular access for blood purification, by which continuous arteriovenous hemofiltration blood purification system was established. Blood flow, substitution fluid flow and ultrafiltration rate were regulated by rotary mini?pumps. Blood purification therapy continued for 4 hours on the basis of maintained anesthesia and effective anticoagulation. The safety of continuous arteriovenous hemofiltration blood purification systems was evaluated by comparing the arterial blood gas, electrolyte indexes and blood glucose during the blood purification therapy. Closely monitoring the vital signs of rats, such as blood pressure and heart rate, and observing whether there were any side effects, such as massive haemorrhage, thrombogenesis and gas embolism in the therapeutic process. Results There were no obvious changes of arterial blood gas, electrolyte indexes and blood glucose during the blood purification therapy (P>0.05). The vital signs did not fluctuate acutely before and after the blood purification therapy (P>0.05). The incidence rate of side effects was very low. Conclusions
Continuous arteriovenous hemofiltration blood purification system had no obvious adverse effects on healthy rats. Our blood purification system for rats appears to be safe and reliable.

Objective To observe the effects of autophagy on the expression of synaptic plasticity related protein, growth-associated pro-tein-43 (GAP-43) and microtubule associated protein-2 (MAP-2), in CA1 area of hippocampus of vascular dementia rats. Methods Nine-ty-six healthy male Sprague-Dawley rats were randomly divided into sham group, vascular dementia model group (VD group), autophagy in-hibitor 3-methyl adenine preconditioning group (3-MA group) and autophagy agonist rapamycin preconditioning group (Rap group). Each group was divided randomly into subgroups of one week, two weeks, four weeks and eight weeks after modeling, six rats in each group. The vascular dementia rat model was established with modified Pulsineli's four-vessel occlusion. The expression of GAP-43 and MAP-2 in CA1 area of hippocampus were detected with immunohistochemistry. Results Compared with the sham group, the expression of GAP-43 protein increased, and the expression of MAP-2 protein decreased at every time point in VD group (P<0.01). Compared with VD group, the expres-sion of both GAP-43 and MAP-2 increased in 3-MA group (P<0.05), and decreased in Rap group (P<0.05). Conclusion Autophagy may in-hibit the expression of synaptic plasticity related protein, GAP-43 and MAP-2, in CA1 area of hippocampus in vascular dementia rats, indi-cating inhibition of autophagy may promote synaptic remodeling.

Objective To explore the clinical value of a loop-mediated isothermal amplification (LAMP) method for detection of Mycobacterium tuberculosis in the application of primary medical institutions . Methods The sputum samples from 140 patients with clinically diagnosed pulmonary tuberculosis detected by LAMP and anti-acid staining method .McNemar test and Kappa test were used for statistical analysis . Results In 140 cases of sputum ,the positive detection rate of anti-acid staining and LAMP method was 29 .3%(41/140)and 45 .0%(63/140) .Compared with the smear method ,the sensitivity of LAMP method was 97 .6%(40/41) ,the specificity was 76 .8%(76/99) .The overall consensus rate was 82 .9%(116/140) .Com-pared with the two methods ,the consistency between the two is moderate and statistically significant (Kappa＝0 .642 ,P＜0 .05) .Compared with the smear method ,the LAMP method used in large-scale specimen examination takes shorter time .Conclusion In sputum sample detection ,positive detection rate of LAMP method is better than antacid staining smear method .It has good batch detection ability and good clinical application prospect .

Objective To explore the influencing factors of the collateral circulation formation in patients with acute cerebral infarction.Methods Three hundred and fifty-two patients with acute cerebral infarction were included in this study,the clinical date of their head and neck 256 slice spiral CT angiography (CTA)examination was analyzed.According to the formation of collateral circulation in the head and neck CTA imaging results,it is divided into the collateral circulation group and the non-collateral circulation group.The clinical data of the two groups were compared.The influencing factors of the formation of collateral circulation in patients with acute cerebral infarction were analyzed by single factor analysis and multivariate logistic regression analysis.Results (1)In 352 cases of acute cerebral infarction,197 cases(56.0%)had collaterals,155 cases (44.0%)had none collateral.(2)Single factor analysis showed that age(t=-2.860,P=0.004),hypertension combined with diabetes(χ2 = 10.709,P= 0.001),history of TIA(χ2 = 4.626,P= 0.034),low density lipoprotein(t=-2.176,P=0.030),high homocysteine(t=2.885,P=0.004),cerebral vascular stenosis(Z=-5.936,P=0.000),posterior circular lesions(χ2=8.548,P=0.004)were the influencing factors in the formation of collateral circulation.(3)Multivariate Logistic regression analysis showed that old age(OR=1.031;95%CI=1.008-1.054;P=0.007),hypertension combined with diabetes(OR= 2.009;95%CI=1.159-3.482;P=0.013),high homocysteine(OR=1.023;95%CI,1.005-1.041;P=0.014),circular lesions(OR=1.727;95%CI=1.063-2.804;P=0.027)were relatively independent risk factors in acute cerebral infarction patients with none circulation,the degree of cerebral vascular stenosis(OR=0.507;95%CI=0.389-0.661;P=0.000),low density lipoprotein(OR=0.723;95%CI=0.532-0.982;P=0.038)served as protective factor.Conclusion Old age,hypertension combined with diabetes,high homocysteine and posterior circulation lesions are risk factors for the formation of collateral circulation in patients with acute cerebral infarction,cerebral vascular stenosis degree and low density lipoprotein can promote the formation of collateral circulation.

Crigler-Najjar syndrome (CNS) is an autosomal recessive disorder in which the content of plasma unconjugated bilirubin is increased due to the reduction or complete deficiency of the activity of bilirubin uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1), classified as CNS type Ⅰ and Ⅱ. CNS type Ⅰ is the most severe, which will develop into kernicterus, damage the brain nervous system, and even threaten the life of patients. This article introduces six CNS treatment techniques, including phototherapy, plasma exchange, drug therapy, liver transplantation, hepatocyte transplantation and gene therapy. The applicable patient types, treatment effects and existing deficiencies of each technique were summarized. Phototherapy, plasma exchange, drug therapy and hepatocyte transplantation can temporarily control serum levels and reduce the risk of jaundice, but cannot completely restore UGT1A1 enzyme activity; liver transplantation is currently the only treatment option for CNS type Ⅰ patients, but is limited by suitable liver donors and post-operative immune rejection. Gene therapy has the most promising application in the treatment of genetic disorders such as CNS, which can provide more viable therapeutic techniques for CNS patients.