Objective To study the role of down regulation of bcl 2 expression of human cholangiocarcinoma cells with bcl 2 mRNA cleaving ribozyme (RZ). Methods Synthetic gene of bcl 2 mRNA cleaving RZ was recombined with the expression vector of eukaryotic cells. The recombined vector was used to transfect human cholangiocarcinoma cells. The expression of bcl 2 in human cholangiocarcinoma cells was observed by immunocytochemistry and flow cytometry. Results The expression of bcl 2 in human cholangiocarcinoma cells transfected with RZ gene was lower than that of control group. Spontaneous apoptosis was observed in a few of the cells. Conclusion bcl 2 mRNA cleaving RZ can down regulate the expression of bcl 2 of cholangiocarcinoma cells notably and can also induce spontaneous apoptosis.

Objective To investigate the incidence of contrast induced nephropathy (CIN) among different patient groups after contrast agent injection.Methods A total of 1243 patients were included in this study (male =694,female =549).The SCr level one week before and 72 hours after the CT examination and the incidence of CIN were recorded and comparison was made among groups according to sex,age,body mass index (BMI),the history of high blood pressure (HBP),diabetes mellitus (DM),chronic kidney disease (CKD),chronic heart failure (CHF),tumor,nephrotoxicity drug (NTD) usage.The frequency,type,dose and injection velocity of the contrast media(CM)were also recorded.Multivariate predictors of CIN were identified by Logistic regression using step-wise selection with entry and exit criteria of P ＜0.10,results were tabulated as odds ratios (OR) with 95％ confidence intervals (CI).Results Among 1243 consecutive patients,the incidence of CIN was 5.5％ (68/1243).Patients with a history of HBP,DM,CHF,CKD or tumor presented with higher incidence of CIN than that of controls (5.9％,51/868 vs.4.5％,17/375).CIN developed in 9 of 203 patients (4.4％,9/203) with CKD and in 59 of 1040 patients (5.7％,59/1040)without CKD.There was no significant difference between the two groups(x2 =0.51,P =0.30).In CKD (-) group,the incidence of CIN was higher in females,patients with DM and patients using LOCM than those of males,DM (-) and using low osmolality contrast medium (IOCM) (P ＜ 0.05),but there was no statistical significance in CKD (+) group.Logistic regression analysis showed that women,age ≥ 75 years,DM,LOCM,NTD,tumor,the time of using CM more than once per month were the most significant predictors of CIN (OR ＞ 1).Conclusion Women,age ≥ 75 years,LOCM,NTD,tumor,and the frequency of using CM more than once per month were more likely to develop CIN.

Objective: To explore the effects of peritoneal dialysis on acute renal insufifciency and on relevant blood indicators in children with congenital heart disease (CHD) after the operation.
Methods: A total of 48 CHD patients received direct open heart surgery by cardiopulmonary bypass and suffered from post-operative acute renal insufifciency in our hospital from 2011-12 to 2014-12 were retrospectively analyzed. The patients were divided into 2 groups: Peritoneal dialysis group and Routine medication group,n=24 in each group. The differences of renal function indexes, the blood levels of electrolyte and inlfammatory factors were compared between 2 groups.
Results: Compared with Routine medication group, the patients in Peritoneal dialysis group presented decreased serum critinine, BUN and urine β2-micro globulin, 24-hour protein level,P<0.05; decreased blood K+ level and increased blood levels of Ca2+, HCO-3P<0.05; decreased plasma levels of hs-CRP, TNF-α, IL-6,P<0.05. The cure rate and mortality in Peritoneal dialysis group were better than those in Control group,P<0.05. No long term complication related to peritoneal dialysis was found.
Conclusion: Peritoneal dialysis may improve the renal function in CHD patients with post-operative acute renal insufifciency and optimize the blood levels of electrolyte and inlfammatory factors.

Objective To investigate the prevalence of BRAF T1799A mutation and RET/PTC rearrangement in Qingdao and detect the expression of platelet-derived growth factor B (PDGF-B) in order to investigate the correlation between gene mutation and PDGF-B.Methods Fresh tissue from 48 papillary thyroid carcinomas (PTC) patients was examined for BRAF mutation RET rearrangements (RET/PTC1 and RET/PTC3) by PCR,followed by direct-sequence analysis.The expression of PDGF was analyzed by immunohistochemistry.Results Among the 48 patients,14 (29.2％) were micro PTC; 18 (37.5％) had BRAF T1799A mutations and 23(47.9％) had RET/PTC rearrangement.There were 17 (35.4％) cases of RET/PTC1 and 6 (12.5％) of RET/PTC3,with no multiple rearrangements.Both BRAF T1799A mutation and RET/PTC rearrangement were present in 6 (12.5％) cases of non-micro PTC.The level of PDGF-B expression in BRAF T1799A positive was higher than that in the negative,and the level of PDGF-B expression in RET/PTC3 was higher than that in RET/PTC1 (P ＜ 0.05).The more advanced neoplasm stage was,the stranger PDGF-B expression was.Conclusions The incidence of BRAF T1799A mutation and RET/PTC rearrangement is higher in Qingdao.BRAF T1799A mutation and RET/PTC3 rearrangement in patients suggests a poorer prognosis than the negative one.The BRAF T1799A mutation and RET/PTC3 rearrangement may strengthen the expression of PDGF-B.Both variations suggest a poor prognosis.

By using 5'-nucleotidase-alkaline phosphatase (5'-Nase-AlPase) double staining method, the fine distribution of the lymph vessels in the stomach wall of the rabbit, guinea pig and rat has been observed. Under light microscope lymph capillaries showed strongly 5'-Nase positive reaction with brown or dark brown staining. However blood capillaries revealed significantly A1Pase activity with blue staining. The lymph capillaries were seen in all layers of the stomach wall in three species, but the lymph vessels were only shown in the submucosa, muscularis and serosa. In the mucosa lymph capillaries could be found in the deep layer of the lamina propria. In the submucosa there were lymph vessels and capillaries. In the muscularis lymph capillaries and vessels not only could be found between oblique and circular muscle or between circular and longitudinal muscle, but also among muscle fiber bundles of each layer. In the serosa there were larger lymph vessels situated near the muscularis. The observation on semithin and ultrathin sections also shown the similarity in the lymph vessels distribution. There was no difference in the distribution of lymph vessels in the stomach wall between rabbit, guinea pig and rat.

Background The use of adaptive optics (AO) system in ophthalmic clinic and basic studies has increased in recent years.However,there are few reports on the stability of ocular aberrations after correction.Objective This study was to analyze the stability of aberration after correction by observing the repeatability of ocular aberration measurements.Methods Forty-one postgraduate school students and volunteers who meet the conditions were included from February to April 2014.The Zernike aberration coefficients including astigmatism (Z2-2,Z22),defocus (Z02),trefoil (Z33,Z3-3),coma (Z3-1,Z13),spherical aberration (Z40) and the value of root mean square (RMS) including 3rd-order to 7th-order aberrations,total higher-order aberrations (HOAs) and total ocular aberrations (TOAs) were measured by using AO system.The repeatability and stability of these data after corrected with AO system were analyzed.The repeatability was evaluated by ANOVA,within-subject standard deviation (Sw),repeatability (r) and intra-class correlation coefficients (ICC).The stability was evaluated by the nonparametric Friedman's rank test.Results AO system showed excellent repeatability on Z2-2,Z22,Z20 and TOA RMS (ICC＞ 0.9),good repeatability on Z13,Z33,Z3-3,Z40,3rd-order RMS,4th-order RMS,HOA RMS (ICC ＞ 0.75),poor repeatability on Z3-1,5th-order RMS,6th-order RMS,7th-order RMS (ICC ＜ 0.75).Repeatability (2.77 Sw) values ranged from 0.009 mm (7th-order RMS) to 0.163 mm (Z31).After low-order ocular aberrations were corrected,It was founded that Z2-2,Z22 reached stable state at the 4th second;Z02 was stable at the 6th second;Z3-3 and Z33 reached stable state at the 4th second and third second,separately;Z13 was stable from 3rd-second to 9th-second,Z3-1 was stable at the 4th-second.Z40 and HOA RMS were stable at the third second and fifth second,respectively.The Z2-2,Z02,Z22,Z3-3,Z3-1,Z33,Z40 and HOA RMS were significantly different among different time points before and after low-order aberrations correction (all at P ＜ 0.05).Z2-2,Z22,Z20 reached stable state at the 4th-second,3rd-second and 5th-second,respectively;Z3-3,Z33 reached stable state at the 2nd-second and 3rd-second,respectively;Z3-1and Z40 reached stable state at the 2nd-second;HOA RMS reached stable state at the 5th-second.Conclusions After correcting the human ocular aberration,different aberrations can reach stable state at different time.The time of Z02,Z22,Z3-3,Z3-1,Z40reaching stable state after 2nd-order to 5th-order ocular aberrations correction was earlier than those of lower-order aberrations correction.

Although tumor treatment models have been continuously improved in clinical practice, cancer remains a serious threat to human health. The effect of probiotics on tumor therapy has received extensive attention. As a common colonizer of the intestinal mucosa, Akkermansia muciniphila(AKK) has a well-defined role in metabolic diseases, but its complex role in tumor development and therapeutic efficacy has not been fully elucidated. The unique properties and physiological roles of AKK play an important role in different solid tumors and it may be a potential biomarker. This article provides a review of previous studies and proposes clinical strategies to influence the abundance of AKK to provide a theoretical reference for the development of next-generation probiotics and the reshaping of the tumor treatment landscape.

Objective:This work aims to detect the levels of miR-200c/141 methylation and miR-200c/141 in gastric cancer tissue and investigate the relationship between miR-200c/141 expression and clinical parameters. Methods:The methylation status of miR-200c/141 CpG island and miR-200c/141 in gastric cancer tissue specimens was evaluated by qRT-PCR or BS-MSP method. We analyzed the relationship among the methylation status of miR-200c/141 CpG island, expression level of miR-200c or miR-141, and clinical parame-ters. Results:The status of miR-200c/141 CpG island methylation in gastric cancer tissue was significantly higher compared with that in paracarcinoma tissue. MiR-200c and miR-141 were markedly decreased in gastric cancer tissue compared with those in adjacent tis-sue. MiR-200c/141 CpG island methylation was negatively related with the expression of miR-200c and miR-141 in gastric cancer speci-mens. Conclusion:The upregulation of miR-200c/141 CpG methylation inhibits miR-200c/141 expression in gastric cancer tissue.

AIM:To observe the effect of Wnt/β-catenin signaling pathway on diabetic ulcer.METHODS:Diabetic animal model was established in the female Wistar rats by intraperitoneal injection of low-dose streptozotocin fol-lowing high-fat diet feeding.A circular wound was made on the dorsum of the rats in both control group and diabetic group. The condition of wound healing was recorded and the structures of the wound tissues were observed by HE staining in the 2 groups at 3, 7 and 14 d after wounding.The expression ofβ-catenin, GSK-3βand Rspo-3 at mRNA and protein levels in the wound tissues was detected by RT-PCR and ELISA.RESULTS:In diabetic group, the wound healing rate was lower (P<0.05), and the inflammatory cells, fibroblast cells and new capillaries in the wound tissues were fewer than those in control group.The expression of β-catenin and Rspo-3 at mRNA and protein levels in the wound tissues in control group was significantly higher than those in diabetic group, and the expression of GSK-3βwas exactly the opposite (P<0.05). CONCLUSION:The down-regulation of Wnt/β-catenin probably resultes from the decreased level of Rspo-3, which may be one of the reasons for delaying the diabetic ulcer healing.

ABSTRACT:To understand the status of contamination and antimicrobial resistance of Staphylococcus aureus (S .aureus) isolated from raw cow milk ,600 samples were collected from five different provinces in China .After isolation and identification of S .aureus from these samples ,157 S .aureus isolates were obtained .Furthermore ,their resistant phenotype and resistant gene were analyzed by MIC detection ,as well as nine drug‐resistance genes determination and analysis .The result revealed that most of the S .aureus isolates contained multiple antibiotic resistant genes ,and all of the isolates were resistant to penicillin . Of 98 .73% isolates were resistant to amoxicillin .Conversely ,all of the isolates were sensitive to enrofloxacin and vancomycin . Moreover ,there were 29 MRSA positive strains among which 22 strains carrying femA gene .In conclusion ,much more atten‐tion should be paid to the drug‐resistance capabilities of field S .aureus isolates in raw cow milk ,and further monitoring of drug resistant S .aureus strains in milk should also be strengthened so as to ensure milk safety .