Objective To construct the eukaryotic expression vector for mouse microRNA miR-21 and identification its expression activ-ity in 293 cells. Methods The genomic sequence containing pre-miR-21 was amplified from mouse genomic DNA by PCR and cloned into the pRC/CMV plasmid. The constructed recombinant plasmid pRC/CMV-mmu-miR-21 was transfected to 293 cells by lipofectamine 2000, and the stably transfected cells were screened with G418,from which total RNA was extracted for detecting the expression of mature miR-21 by northern blot. In the meantime,a luciferase report plasmid examing the activity of miR-21 named pmiR-21-Luc reporter was also construc-ted,and luciferase activity analysis indicated the product of pRC/CMV-mmu-miR-21 indeed had biological activity. Results Both restriction enzyme digestion analysis and sequencing proved the recombinant plasmids were constructed correctly. The miR-21 was highly expressed in the screened clones of 293 cells and it had good biological activity. Conclusion The eukaryotic expression plasmid of mouse miR-21 was successfully constructed,which laid the foundation of further investigation of the role of miR-21 during skin wound healing.

Objective ToinvestigatethementaldevelopmentsituationofpreschoolchildreninTaiyuan. Methods 1174preschoolchildrenoftwourbanareasinTaiyuanwereselected.TheyweretestedwithDenverDevel-opmentScreenTest(DDST).Results Thepassingratewas94.80%in1174children.Theirindividual-social reaction ability and movement ability were good in the three groups.In the subject-touch reaction ability,the passing rate ofvertical tilt at 30°insidewas 36.36%in 4 years old group,which was higher than the other projects.In the speaking ability,the passing rates of three right in four problems with knowing preposition were 15.88% and 27. 22% in 4 and 5 years old group,which were higher than the other projects.The culture degree of the mental retar-dation children's parents were lower than those of the normal children's parents,the difference were statistically signifi-cant(χ2=4.485,P=0.034;χ2=7.577,P=0.006).Conclusion DDSTisaquicklyscreeningmethodformental retardation children,it is suggested to be used in medical and health examination for preschool children.

Objective To explore the feasibility of using portable γ spectrometer to rapidly identify the low activity of depleted uranium and the underlying conditions.Methods Firstly,high purity germanium (Ge) γ spectrometer was used to analyze energy spectrum of DU samples (5 g) and calculate nuclide percentage of 235U in an attempt to ascertain the properties of these DU samples.The portable γ spectrometer was used to provide the evidences for identification of DU samples.Secondly,portable γ spectrometer was also used to identify DU samples of same group.These samples contain 1 g DU powder and 0-5 g environmental clay powder,which were sealed with double layer pocket,and then detected with a distance of 1-5 cm during the longest detection time of 10 min.According to the detection of nuclide activity of 238U and 235U in the samples and the subsequent calculation of specific activity,the nuclide percentage composition was calculated and the existence of DU was confirmed if this value of 235U was less than 0.718％.Results The activity of 238U was detected using portable γ spectrometer under all test conditions,while the activity of 235U was detectable only under certain test conditions (MA ≥ 1 g,DN ≤ 1 cm).Under the condition that the 238U and 235U was both detected,the nuclide percentage of 235U was all less than 0.718％,which suggested that the DU was confirmed.Conclusions The energy spectrum of low activity of DU and the type of nuclide could rapidly be identified and evaluated by using portable γ spectrometer.This is same as the conclusions obtained with high purity Ge γ spectrometer,α spectrometer and ICP-MS.

Objective To explore the effect of long-term depleted uranium (DU)ingestion on testosterone production in rats, and its involvement mechanism. Methods Male and female rats (F0 and F1 respectively) for 160 days, respectively. The contents of testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) in serum were detected in 20 months of F0 generations, and 15 months of F1 generations. RT-PCR was used to analyze the levels of StAR mRNA and P450scc mRNA. Results Compared with the normal control group, the testosterone contents in exposed F0 and F1 generations increased, the lowest was 51.73 U/L, but those of LH and FSH decreased. The expression of StAR mRNA in the low-doze group of F1 generation (StAR/β-actin = 1.35) was up-regulated, down-regulated for other groups.compared with the normal control group (P450scc/β-actin = 0. 313), the expression of P450scc mRNA in the low- and high-dose groups of F0 generation were decreased (P450scc/β-actin = 0.21), and those in the low- and high-dose groups of F1generation were increased (P450scc/β-actin = 0.623) (P ≤ 0.01). Conclusion Long-term DU exposure inhibit the male reproduction by intervening the sexual hormone production through down-regulated the expression of StAR mRNA and P450scc roRNA.

Objective To compare the expression of nitric oxide synthase (NOS ) in patients with gastroesophageal reflex disease (GERD)and healthy controls.Methods The distribution and relative protein amount of two NOS subtypes (nNOS and iNOS)were determined with immunohistological method,and their mRNA levels were measured with real-time PCR method.Results The nNOS and iNOS were mostly distributed in the cytoplasm in epithelia of esophageal mucosa.The nNOS and iNOS in reflux esophagitis (RE)were significantly higher than in non-erosive reflux disease (NERD)patients and healthy controls (P <0.05 ).The mRNA levels of nNOS and iNOS were also significantly higher in RE patients than in NERD patients and healthy controls (P <0.05).Conclusion The expressions of nNOS and iNOS were increased in the esophagus of RE patients,which may be related to the effects of NO on the onset of GERD.

objective To compare three risk prediction system,the physiological and operative severity score for the enumeration of mortality and morbidity(POSSUM),the Portsmouth POSSUM (P-POSSUM)and the colorectal POSSUM(Cr-POSSUM)for the accuracy in predicting operative mortality of patients of colorectal cancer in a single Chinese referral hospital setting. Methods Data of 903 patients,who undergone surgery for colon and rectal cancers from 1992 to 2005 at Peking University Third Hospital,were enrolled in the study.POSSUM,P-POSSUM and Cr-POSSUM was used respectively to predict the mortality rate.ROC curve was applied to judge the differentiation ability of each score.Model goodness-or-fit was tested by the Hosmer-Lemeshow statistic and subgroup analysis was performed by the ratio of observed to expected deaths(O∶E ratio). Results The actual inhospital mortality in our series was 1.0%(9/903).The oredicted mortality rate by POSSUM,P-POSSUM and Cr-POSSUM were 5.6%,2.8% and 4.8%respectively.These predicted mortality rate were significantly higher than actual mortality of our patients.The O∶E ratio was 0.18,0.35 and 0.2 respectively. Conclusion The predicted mortality rate of POSSUM,P-POSSUM and Cr-POSSUM were significantly higher than actual observed mortality rate in a single Chinese referral hospital for patients of colorectal cancer.

Objective To evaluate a score system(Association of Coloproctology of Great Britain and Ireland ACPGBI)in prediction of postoperative mortality from colorectal cancer patients in a Chinese hospital. Methods We analyzed retrospectively 904 patients with histologically confirmed colorectal cancer who had colorectal surgery from 1992 to 2005.There were 525 colonic cancer patients and 379 rectal cancer patients.We divided patients into several groups according to operative urgency(elective or emergency);surgeons(colorectal specialists or other surgeons);cancer location(colon or rectal).According to ACPGBI score we got the prediction.This prediction was compared with the actual mortality;Chi-square test,receiver operator characteristic curve(ROC),Hosmer-Lemeshow goodness-of-fit test were used.Results Observed overall mortality within 30 days after surgery was 1.0%(9/904),and the predicted mortality was 8.3%(75/904).In all the subgroups the predicted momdity wag higher than observed mortality.We found that the actual mortality was higher in an individual subgroup in which the predicted mortality was higher. Conclusions For colorectal cancer patients undergoing a surgery the predicted mortality of ACPGBI score system was higher than the actual mortality in a Chinese hospital.

Objective To find out the best method for elimination of uranium contamination in rat tissues as low as possible by removal of shrapnel fragments. Methods Experimental rats were divided into six groups: route group, decontamination before surgery group, decontamination in incision group, changing surgical appliances group, removing tissues around group, and comprehensive method group. Uranium concentrations in tissues and fluids in all groups were measured at 7, 14, and 21 d after operation. The efficiency of decontamination by different methods was compared. Results The highest uranium concentration in tissues was found in the route group, but the lowest in the comprehensive method group, and the second lowest in removing tissues around group. Conclusion The comprehensive method is the best one in all of the surgical removal methods. The soft tissues around DU shrapnels should be removed if they are not critical organs.

OBJECTIVESTo evaluate the characteristic morphology of heel spur, and to investigate the relationship of heel spur and plantar heel pain.

METHODSFrom June 2005 to April 2009, 210 cases (254 feet) with heel spur (according to Denis Pain Scale) were divided into cases group 1 (P2, n = 46), 2 (P3, n = 44), 3 (P4, n = 42), 4 (P5, n = 36) and controls group (P1, n = 42). Three-dimensional reconstruction of heel spur was performed in all groups using volume rendering based on multi-slice CT data by Super Image orthopedics edition 1.0. The characteristic morphology of heel spur was observed and the data were measured and analyzed, involving the width of basilar part, the length, the angle between heel spur and planta pedis, and the angle between the longitudinal axis of calcaneus and heel spur.

RESULTSParts of cases groups displayed coarse arcuate edge and undersurface with one or more little heel spurs adhere to heel spur, of which the numbers were greater than controls group, especially in cases group 4. No significant difference of the width of basilar part of heel spur was found among 5 groups (F = 2.32, P > 0.05). However, obvious difference was found in the length, the angle between heel spur and planta pedis, and the angle between the longitudinal axis of calcaneus and heel spur (F = 8.23, 6.82, 5.87, P < 0.05). Compared with the controls group, the angle between heel spur and planta pedis of cases groups had higher degrees, but the difference of the other data presented irregular.

CONCLUSIONSThe characteristic morphology of heel spur varies in patients associated with plantar heel pain. No correlation is found between the severity and the morphological data, including the width of basilar part, the length, the angle between heel spur and planta pedis, and the angle between the longitudinal axis of calcaneus and heel spur.

Aged ; Calcaneus ; diagnostic imaging ; pathology ; Case-Control Studies ; Female ; Heel Spur ; complications ; diagnostic imaging ; pathology ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Tomography, X-Ray Computed

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.