Objective: To optimize and evaluate the modified RNA-tailing and primer-extension RT-PCR method in relative quantification of microRNAs (miRNAs) in several kinds of tissues. Methods: Small-sized RNAs (

Objective To understand the expression of heat shock protein(HSP70) mRNA induced by avermectins in the hepatic tissue of rats.Methods Wistar rats aged 6 weeks were randomly divided into 4 groups,12 in each including 6 male and 6 female.The rats in the experimental groups were treated with avermectins by gavage once a day at the doses of 2.5,1.25 and 0.625 mg/kg respectively for 14 consecutive days,one group was the negative control.The general state was concerned and after the exposure,the content of total protein(TP) in the serum was determined,the hepatic tissue was sampled to examine the expression of HSP70 mRNA by polymerase chain reaction(PCR).Results The content of TP in the serum in the 2.5 mg/kg group was lower than that in the control group(P

Objective To explore the risk factors for complicated hypertension in type 2 diabetic patients. Methods A community-based case-control study was conducted and some risk factors of complicated hypertension in type 2 diabetic patients were analyzed.126 cases and 273 controls were type 2 diabetic patients with and without hypertension respectively.Results The results from univariate logistic regression analysis showed that the occurrence of complicated hypertension in type 2 diabetic patients was statistically related to the body mass index(BMI),waist hip ratio(WHR) and the intake of grain crop.The corresponding OR with 95% CI were 1.474(1.101~1.973),1.478(1.180~1.852) and 1.349(1.098~1.657),respectively.After adjusted for confounding factors,the associations mentioned above were still statistically significant.No associations were observed between complicated hypertension in type 2 diabetes and diabetes duration,glycemia control,exercise,smoking and alcohol drinking,serum total cholesterol,triglycerides,high density lipoprotein,low density lipoprotein and very low density lipoprotein cholesterol levels.Conclusion The high body mass index(BMI),high waist hip ratio(WHR) and high intake of grain crop may increase the risk for the complicated hypertension in type 2 diabetic patients.

Neonatal cerebral infarction is an area of damaged cerebral tissue resulting either from disruption to blood flow in a major cerebral artery from thrombosis or embolism or from thrombosis in a major cerebral vein.The pathogenesis is unknown at present,many studies have shown that genetic,mother hypertension,gestational diabetes,smoking,neonatal congenital heart disease,infections,meningitis are the risk factors of neonatal cerebral infarction.

Objective: To investigate the non-specific and inaccurate amplification in cases of highly similar sequences among family members and the length heterogeneity of mature microRNA ( miRNA) ,and find a condition that discriminates maximally among similar miRNA family members and detects the accurate expression level of miRNAs. Methods: Primers with their mismatches and/or 3' end at different positions were designed. Amplification efficiencies were compared using matched and various mismatched primers by RNA-tailing and primer-extension RT-PCR at different annealing temperatures. Expression levels of several miRNAs in mouse brain were compared using miRNA specific primers with different termini. Results: Raising annealing temperatures 12℃-14℃above the T_m of the primers maximally increased amplification specificity without sacrificing sensitivity. Primers designed with their termini on or near variant positions could efficiently discriminate between miRNA isoforms. Using primers that terminated before the end of the mature miRNA did not miss the detection of shorter mature miRNA and provided accurate expression levels. Conclusion: Careful primer design and higher annealing temperature can increase specificity and accuracy of real time PCR miRNA detection.

Objective:To examine global expression levels of microRNAs(miRNAs) in mouse cerebrum and to provide an important basis for detailed studies of individual miRNAs,their target genes,the miRNA-related regulatory networks in the mammalian central nervous system,and their implications in diseases.Methods:Low molecular weight RNA from cerebrum of five C57BL/6J mice were tailed and reverse transcribed by extended RT-primer.miRNA primers were carefully designed and arrayed on plates according to the Tm of each primer.PCR was carried out at different annealing temperatures using a gradient real-time PCR instrument.The relative expression level of each miRNA was calculated using 5sRNA for normalization.Results:Among the 285 miRNAs detected,260 were positive with varying abundance.Their frequency distribution was approximately a normal distribution.The expression levels of most miRNAs were in accordance with previously published results by microarray.However,the positive rate was higher than that detected by microarray.miRNAs originating from the same hairpin precursors expressed at similar or significantly different levels.Clusters of proximal miRNAs were similar or quite different in abundance.It is suggested that the fate of miRNA after transcription determined their abundance.Conclusion:Using the RNA-tailing and primer-extension PCR array method,we obtained expression profile of miRNA in mouse cerebrum,especially the relative expression data of many low abundant miRNA in mouse cerebrum,which will be of special help for studying the fine-tuning function of low-level miRNAs.

Objective To investigate the effects of human milk fortifier(HMF)addition at different time points on the growth,development and the incidence of complications in very low birth weight(VLBW)infants.Methods A total of 93 VLBW infants admitted into Neonatal Intensive Care Unit of Tianjin Central Hospital of Obste-trics and Gynecology from January to September 2015 with more than 80％of total milk intake during hospitalization,excluding those who had severe asphyxia or abandoned treatment and died,were collected.The included cases were divided into 2 groups by using completely randomized grouping method,early fortification group(n=48)and delayed fortification group(n=45)adding HMF with the enteral intake of 50 mL/(kg·d)and 100 mL/(kg·d),respectively.The outcomes included growth development and the incidence of complications during hospitalization.Then,t test and chi-square test of independent samples were used for statistical analysis.Results There was significant difference in the weight growth rate between the 2 groups,and the growth rate of early fortification group and delayed fortification group were(15.4±2.4)g/(kg·d)and(13.6±2.3)g/(kg·d),respectively(t=3.043,P=0.004).There was no significant difference in height growth rate,head circumference growth rate,weight at 34 weeks postmenstrual age,time of recovering birth weight and parenteral nutrition,hospitalization duration,body weight,body length,head circumference at discharge and the incidence of extrauterine growth retardation between the 2 groups(all P>0.05).There was no statistical difference in incidence of feeding intolerance,necrotizing enterocolitis,nosocomial infection,retinopathy of prematurity,bronchopulmonary dysplasia between the 2 groups(all P>0.05).Conclusions HMF with enteral intake of 50 mL/(kg·d)contributes to weight gain rate in VLBW infants during hospitalization,but not to the increase in the incidence of complications.

In order to explore the effects of diet and nutritional factors on breast cancer incidence, a case-control study was conducted in Tianjin. The study consisted of 244 cases diagnosed between September 1986 and September 1987 and equal number of matched controls from patients other than breast cancer in general hospital.The socio-demographic data, histories of menstruation and reproduction etc were collected by interview. The interview on diet was conducted by recalling the frequencies and amounts of various foods eaten one year prior to the diagnosis' of their current diseases. Data were analysed by using a nutrient-estimate program and conditional logistic regression model, chi square test and u-test.After adjusting for confounding factors, the results showed that high fat, high calorie, high monounsaturated fatty acid and low vitamin C intakes were risk factors for female breast cancer. High fat percentage of energy and low vegetable intake were associated with increased risk for breast cancer.

Objective To evaluate the efficacy of balloon dilatation in the treatment of intrahepatic biliary strictures in patients with liver transplantation. Methods Of the 100 patients with liver transplantation, 16 patients had intrahepatic biliary strictures and received balloon dilatation treatment. Results Initial technical balloon dilatation was successful in 14 cases but failed in 2 cases. There were no procedure-related complications. 4 restenosis occurred and they were treated with repeated balloon dilatation treatment. Conclusion Balloon dilatation represented an effective and relatively safe treatment for biliary stricture in liver transplant recipients. For restenosis, balloon dilatation was also an effective treatment.

OBJECTIVE: To investigate the effects of combined inhibition of signal transducer and activator of transcription 3 (STAT3) and hypoxia-inducible factor-1α (HIF-1α) in the enhancement of chemosensitivity of the model of human laryngeal squamous cacinoma in nude mice. METHOD: Model nude mice were divided into six groups randomly: control group(A) , cisplatin group(B) , cisplatin and AG490 group(C) , cisplatin and HIF-1α⁻/⁻ group (D), cisplatin combined AG490 and HIF-1α⁻/⁻ group (E), HIF-1α⁻/⁻ group (F) (only in calculating tumor inhibition rate). 3mg/kg cisplatin was administered by peritoneal injection for 3 days. Then cisplatin and 10 mg/kg AG490 were administered every other day for 12 days. The expression of Ki67 and HIF-1α was detected by immunocytochemical method. Western blot was used to detect the expression of p-STAT3. RESULT: The expression of HIF-1α in group C and group D were lower than that in group B, and there were significant difference respectively (t₁ = 2.782, t₂ = 3.873, P < 0.05); The expression of HIF-1α in group E was lower than that in group C and group D respectively, and there were significant difference respectively (t₁ = 6.140, t₂ = 3.667, P < 0.01). The expression level of p-STAT3 in group C was markedly lower compared with that in group B, and there were significant difference between them (t = 17.840, P < 0.01); There were no difference between the expression level of p-STAT3 in group D and that in group B (t = 0.038, P > 0.05); The expression level of p-STAT3 in group E was significantly lower compared with that in group C and group D respectively (P < 0.01). Tumor inhibition rate of group E was higher than that in group B, group C , as well as group D respectively and there were significant difference respectively (t₁ = 5.509, P < 0.01; t₂ = 3.422, P < 0.05; t₃ = 2.718, P < 0.05 ). Ki67 index of group E was lower than that in group B, group C as well as group D respectively and there were significant difference respectively(t₁ = 8.307, P < 0.01; t₂ = 3.736, P < 0.05; t₃ = 4.524, P < 0.01). CONCLUSION Combined inhibition of STAT3 and HIF-1α could enhance chemo-sensitivity in the model of human laryngeal squamous cacinoma in nude mice.
Animals ; Antineoplastic Agents ; pharmacology ; Carcinoma, Squamous Cell ; drug therapy ; metabolism ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Ki-67 Antigen ; metabolism ; Laryngeal Neoplasms ; drug therapy ; metabolism ; Mice ; Mice, Nude ; Neoplasms, Experimental ; drug therapy ; metabolism ; STAT3 Transcription Factor ; genetics ; metabolism ; Tyrphostins ; pharmacology