Objective To explore origin recognition complex 1(ORC1) expression in the rat vascular muscle cells at different phases of proliferation.Methods Vascular smooth muscle cells(VSMCs) of thoracic aorta of rats in primary culture were obtained by the adherence method of tissue culture.Total RNA of VSMCs was extracted.The expression of ORC1 mRNA of VSMCs at different phases of proliferation was determined by reverse transcription polymerase chain reaction(RT-PCR) and the expression of ORC1 protein by immunocytochemistry and laser confocal microscopy.Results Cultured VSMCs were confirmed by light microscope and immunocytochemistry.The expression of ORC1 mRNA in the quiescence stage of VSMCs was not found.After VSMCs were stimulated with serum,the level of ORC1mRNA had an obvious increase at 6 h,peaked at 12 to 24 h and decreased in the following 24 h.The expression of ORC1 protein was also not found in the quiescence stage of VSMCs,but the level of ORC1 protein during proliferation of VSMCs was significantly increased.Conclusion ORC1 may have an important role during the process of VSMCs proliferation in rats.

Objective To explore the expression of origin recognition complex1(ORC1) during DNA replication progress of rat vascular muscle cells(VSMCs).Methods VSMCs of rat thoracic aorta were obtained by the adherence method of tissue culture.The growth curve was drawn by MTT.The association between DNA replication and the expression of ORC1 mRNA and protein in different growth phases of VSMCs was analyzed.Results The expression of ORC1 mRNA and protein in quiescence stage of VSMCs was not found.After stimulated with serum,the expression of ORC1 mRNA in rat VSMCs increased significantly,peaked at 12-24 h.The expression of ORC1 protein was similar to ORC1 mRNA in VSMCs.Meanwhile,the higher DNA replication of stimulated VSMCs was observed,peaked at 12-24 h after serum addition. Conclusion ORC1 may be involved in the DNA replication of rat VSMCs during the progress of proliferation.

Objective To investigate the influence of RNA interference targeting ORC1 gene on the proliferation of rat vascular smooth muscle cells(VSMCss).Methods VSMCss were transfected with siRNA targeting ORC1 gene by liposome.The expression of ORC1 protein was detected by Western blotting.MTT test and ~(3)H thymidine~()(~(3)H-TdR) incorporation were used to detect VSMCss proliferation.The expression of proliferating cell nuclear antigen(PCNA) in VSMCss was detected by immunocytochemistry.Results After transfected of the three pairs of siRNA targeting ORC1 genes respectively,the expression of ORC1 was all lower than that in the control group(non-transfection and negative siRNA).Especially,that in the group transfected of the second pair positive siRNA decreased most significantly.The optical density of MTT,~(3)H thymidine incorporation and the expression of PCNA decreased significantly in VSMCss transfected with siRNA targeting ORC1 gene as compared with that in the control group.Conclusion ORC1 gene silenced by RNA interference can inhibit VSMCss proliferation.

Objective To study the transfection efficiency, protein expression, and effect of adenovirus-mediated transfection on microvascular endothelial cells transfected by angiopoietin-related protein 2(Ad. ARP2)gene. Methods Mice coronary microvascular endothelial cells(CMECs) were isolated, cultured and transferred by Ad-ARP2. The transfection efficiency and cellular toxicity of adenovirus vector to CMECs were detected by immunofluorescence staining. Expression of Ad. ARP2 in CMECs and the secreted materials in culture medium were measured by Western blot and ELISA and compared among groups of Ad. ARP2, Ad. null, and PBS control. Vascular endothelial cells were incubated with conditional medium containing secreted ARP2, and effects on cells sprouting were observed in matrigel. Results Adenovirus-transfected CMECs showed a very high efficiency. When multiplicity of infection (MOD was 200, the transfection efficiency was 93. 5% ,and no harmful effect on CMECs growth was found. When CMECs were transfected with Ad. ARP2, there was a high ARP2 expression, which was significantly different from that with Ad. Null or PBS. The conditional medium containing ARP2 had an excellent ability to stimulate sprout of CMECs which phenomenon could not be seen in the control groups. Conclusions Adenovirus vector can be transferred into CMECs efficiently and safely. Ad. ARP2 gene transfection allows a high transient expression, and the expression products can stimulate the sprout of microvascular endothelial cells in vitro very well.

The performance of transjugular intrahepatic portosystemic shunt (TIPS) has two key procedures: (1) portal vein branch puncturing, and (2) the correct judgment of the safety of the puncture site. The portal vein branch puncturing is the most important and difficult step for a successful TIPS procedure. Therefore, to find and to establish an proper access to the portal vein is critical. Nowadays, in clinical practice several imaging techniques have been used to localize the portal vein, such as magnetic resonance imaging, sonography, fluoroscopy, arteriography and computed tomography. This article aims to make a general review on these invasive and non - invasive localization techniques when a successful performance of TIPS is expected.

Objective To study the effect of chest wall vibration therapy on bronchiolitis. Methods A total of 64 patients with bronchiolitis were divided into an experimental group and a control group, the former included 34 cases and the latter included 30 cases. The experimental group received both routine treatment and chest wall vi- bration, while the control group only received routine treatment. PaO_2, PaCO_2, SaO_2, Heart Rate (HR) and Respi- ration (R) were observed, respectively, in the experimental group and the control group at the beginning and the end of the third day. Time needed for expectoration and length of hospital stay in the two groups were observed. Results It was shown that PaO_2, PaCO_2, SaO_2 , HR, R were significantly improved at the end of the third day when compared with those at the beginning in both groups(P

To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

Anal Canal/surgery* ; Anorectal Malformations ; Constriction ; Fecal Incontinence ; Humans ; Rectal Prolapse

Objective To summarize the clinical features, diagnosis and treatment of autoimmune pancreatitis (AIP). Methods From March 2003 to January 2008, a total of 16 cases of AIP were reviewed retrospectively. Results The ratio of male: female was 15:1, with a mean age of 61 years old (range:47-79 years old). Jaundice was the main clinical presentation in 81.2% patients. 68.8% patients presented with high serum gammaglobulin, while 66.7% with high serum IgG, 56.2% with elevated ESR, 50.0% with positive rheumatoid factor(RF), 43.7% with eosinophilia, 26.7% with positive antinuclear antigen(ANA),31.2% with elevated lipase, 18.7% with elevated amylase, and 25.0% with elevated CA19-9. 93.7%patients showed diffuse swelling of the pancreas on CT and/or endoscopic ultrasound. Stricture of the main pancreatic duct was seen in 100% patients. Distal common bile duct stricture was seen in 87.5%, while thickened wall of bile duct was seen in 50%. Histological findings of the pancreas EUS-FNA showed nonspecific results in one patient, while no tumor cell was detected in other 5 patients; lymphocytes infiltration was noted in 3 patients; pancreatic fibrosis was seen in 2 patients. 75.0% patients was found to have diabetes or abnormal sugar tolerance, enlargement of the celiac lymph nodes in 43.7%, splenic vein or inferior cava vein involvement in 42.9%, swelling of the maxillary glands in 18.7%, the lacrimal glands in 12%. Prednisone was given to 11 patients, among them 5 patients underwent endoscopic stent placement, and 10 patients responded well while 1 patient discontinued therapy due to intolerance. 2 patients underwent endoscopic stent placement alone and jaundice disappeared. 3 patients received conventional medical treatment. Steroid therapy exerted different effects on levels of the blood glucose, the enlarged maxillary and lacrimal glands improved after steroid therapy. Conclusions AIP occurred in middle aged and senior male predominantly, painless obstructive jaundice was the main clinical presentation, and patients may be accompanied with elevated levels of IgG, hypergammaglobulin, positive RF and ANA, diffuse or focal pancreatic enlargement, pancreatic duct stricture and distal common bile duct stricture. Stent placement could improve the symptoms, and steroid therapy was effective.

Objective To understand the characteristics of cervix diseases in female medical staff and provide basis for effective prevention and control.Methods The results of cervical cancer screening in 1 471 female medical staff in our hospitals in 2008 were compared with the 1 706 ones in 2010.Results Morbidity rates of various cervix diseases were 37.9％ (558/1 471) in 2008 and 50.6％ (864/1 706) in 2010,respectively.Most of them were diagnosed with chronic cervicitis (36.8％ in 2008 and 47.1％ in 2010).The difference of positive rate of screening test (TCT results ≥ASC-US) was significant between those in 2008( 1.1％,16/1 471 )and those in 2010(3.7％,63/1 706) ( x2 =12.65,P ＜ 0.01 ).The results also showed that the incidences of various cervix diseases were related to age,and that high rates of cervicitis and cervical cancer were observed in the middle-aged (35 -50 years old).Conclusions Positive rates of cervical cancer screening test tend to rise,and the health condition of the middle-aged women (35 -50 years old) should be paid particular attention.