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 determine the biological reference interval of urinary conductivity in healthy people and to study the relationship between urine conductivity and other parameters as well as its clinical feasibility.Methods Conductivities of midstream urine specimen from healthy people (n=10119,5074 males and 5045 females,aged under 96 years) or patients with different diseases (n=3449) were determined.Among them the following parameters:conductivity,osmolality,specific gravity,creatinine,urea,uric acid,sodium,potassium and chloride of 250 random urine specimens were simultaneously determined.Results The urinary conductivities in healthy people exhibited normal distribution and significant differences were found in the subjects with different age and sex.The reference range of urinary conductivity was between 10.42?4.61 mS/ch and 24.10?6.81 mS/ch in healthy people and between 7.95 mS/ch?2.40 and 18.01?5.90 mS/ch in the patients with different diseases.Conductivity determined was positively correlated with osmolality (r:0.894),specific gravity (r:0.727),sodium (r:0.698),potassium (r:0.563),chloride (r:758),uric acid (r:0.521),urea (r:0.556) and creatinine (r:0.495).Conclusions Urine conductivity,which determination is simple and rapid,may reflect the conductive capacity of electrolytes in urine and positively correlated with osmolality,so it can be used as a new parameter for urinalysis to diagnose renal concentrative function in routine laboratory.

Objective To study the characteristics and regularities of somatotype growth of Yugu adolescents.Methods The somato-type growth of 989 Yugu adolescents(male:512,female:477)in Sunan was evaluated by the Heath-Carter method.Results The average somatotype of YuGu adolescents in males was mesomorphic ectomorph(3.0-3.6-3.7),and in females,the average somatotype was ectomorphic endomorph(3.8-2.9-3.6).The somatotypes develop from central,endomorphic ectomorph to mesomorphic ectomorph in the male,however,in the female from central,ectomorphic endomorph,endomorphic ectomorph,to mesomorphic endomorph.Conclusion The somatotypes of Yugu adolescents are very different between males and females.In the male group,the somatotypes of the 7-12 year-old group of Yugu adolescents are similar to the Mongolia,Han ethnic,Zhuang ethnic and Hungary.The somatotypes of 13-17 year-old group are similar to Tibetan,Zhuang ethnic,Han ethnic and Daur.However for the female group,the somatotypes of the 7-9 year-old group are similar to Hungary,and the 10-17 year-old group are similar to Tibetan,Zhuang ethnic,Han ethnic and Finn.

Objective: To study the immunotherapeutic effect on the esopgageal adenocarcinoma mediated by gp96-peptide complexes isolated from the same kind of tumor. Methods: gp96- peptide complexes were purified from nude mice tumors burdened by subcutaneous injection of human esophageal adenocarcinoma cell line SEG-1 . gp96-peptide complexes were carried by the dendritic cells(DC) induced from human peripheral blood mononuclear cells to prepare gp96-DC vaccine. The proliferation of lymphocytes was tested with trypan-blue stain. The quantity of interferon-?(IFN-?) released from cytotoxic T lymphocytes (CTL) was detected with ELISA method. The killing effect of CTL on target cell SEG-1 was measured with MTT. Results: We obtained 120 ?g gp96 from 55 g tumor tissue. DC, gp96, and gp96-DC all could elicit the proliferation of lymphocytes and make them becoming into CTL which released IFN-? and showed different degrees of killing effect on target cell SEG-1. gp96-DC has the strongest eliciting effect among them. At the ratio of E(effect) to T(target) as 40∶1,the killing rate was 68%.No significant difference between the effects of CTL induced by DC alone and of lymphocytes without specific antigen on SEG-1 and K562 cells. Conclusion: The gp96-peptide complexes from tumors can improve the effect of eliciting lymphocyte proliferation of DC and make the lymphocyte becoming into CTL more effectively.These CTLs show prominent killing effect on the target tumor cells.

ObjectiveTo establish the proper review rules for the microscopic screening of urine samples tested by automatic urinalysis work station which is composed of LabUMat urine dry chemical analyzer and Urised urine sedimental analyzer.Methods The paired comparison was used to analyze the results tested by microscopy and Urised.A total of 2015 random urine samples were enrolled to establish and validate review rules.All the samples were collected from the inpatients and ontpatients of General Hospital of the People's Liberation Army from May to November 2011 and tested by urinalysis work station.2015 urine samples were firstly tested by urinalysis work station,including both urine dry chemical analyzer and urine sediments analyzer.Then each urine sample was examined microscopically by two technicians-in-charge using double-blind method.The average results from the two technicians were used as review results.Compared with review results,we set up the review rules and evaluated the Irue positive rate,false positive rate,true negative rate,false negative rate (omission diagnostic rate) and review rate.According to different test methods by automatic urinalysis work station,four microscopic review protocols were defined:( 1 ) Protocol 1:based on chemistry results only,microscopy review was performed when any of WBC,RBC,PRO and NIT was positive; (2) Protocol 2:based on urine sedimental analysis only,microscopy review was performed when any of WBC,RBC and CAST count was over upper limit of the reference range ; (3) Protocol 3:if any of BLD vs.RBC,LEU vs.WBC was different between two systems,or quantitative results had two or more than two gradient differences,microscopy review was performed; (4) Protocol 4:if any of BLD vs.RBC,LEU vs.WBC was different between two systems,or CAST was over upper limit of the reference range,or alam appeared,microscopic review was performed.300 randomly selected urine samples were tested to validate the review rules.Omission diagnostic rate and review rate were used to evaluate the rules.Results According to our review rules,the positive samples rate was 41.14％ (829/2015) and the negative rate was 58.86％ ( 1186/2015 ) ; Positive samples were composed of RBC (50.30％),WBC (53.32％) and CAST (3.74％).The review rates of four protocols were 42.93％ (865/2015),39.70％ (810/2015),29.58％(596/2015),18.91％ (381/2015 ),respectively.The false negative rates (omission diagnostic rates) were 6.36％ (128/2015),4.42％ (89/2015),1.34％ (27/2015)and 1.04％ (21/2015)respectively.Protocol 4 was selected as an ideal plan.Additional 300 urine samples were tested using protocol 4 in order to confirm the review rule.The review rate,consistency rate,true positive rate,false positive rate,true negative rate,omission diagnostic rate were 19.67％ (59/300),91.67％ (275/300),35.67％ (107/300),7.67％(23/300),56.00％ (168/300),0.67％ (2/300),respectively.After image review revised,the review rate was 8.67％ (26/300).ConclusionThe review rules established by our research for Urinalysis Work Station can find the abnormal urine samples effectively and exactly and can reduce the workload significantly.(Chin J Lab Med,2012,35:810-814)

Objective To establish the proper review rules for the microscopic screening of urine analyzed by UF-1000i automatic urinalysis work station (composed of UF-1000i urine flow cytometer and AX-4030 urine dry chemical analyzer).Methods A total of 2 839 random urine samples were collected at Chinese People′s Liberation Army General Hospital from September 2009 to February 2010, and were analyzed using UF-1000i urinalysis work station.The parameters obtained from UF-1000i and AX-4030 included RBC, WBC, CAST and ERY, LEU, PRO.After analysis by urinalysis work station, each urine sample was examined microscopically by two technologists using double-blind method.The average results got from the two technologists were regarded as the judging criterion.Based on the criterion, the review rules for the 2 839 urine samples tested by urinalysis work station were created and adjusted, and the true positive rate, false positive rate, true negative rate, false negative rate and review rate of these review rules were calculated.After that, 299 randomly selected urine samples were tested to validate these review rules.Omission diagnostic rate and review rate were used to assess the clinical practicability of the review rules.Results Thirty seven rules for microscopic review and twenty seven rules without further microscopic examination were set up based on six parameters using UriAccess 3.0 Software.The microscopic examination result was taken as the judging criterion, the consistency rate of these rules was 81.11%(2 311/2 839), the true positive rate was 40.51%(1 150/2 839), the false positive rate was 16.17%(459/2 839), the true negative rate was 41.00%(1 164/2 839), the false negative rate(omission diagnostic rate) was 2.43%(69/2 839) and the review rate was 18.28% (519/2 839).Additional 299 urine samples were assayed using UriAccess3.0 software to further verify these review rules.The consistency rate was 82.27%(246/299), the true positive rate was 36.12%(108/299), the false positive rate was 16.39%(49/299), the true negative rate was 46.15%(138/299), the false negative rate(omission diagnostic rate) was 1.34%(4/299), the review rate was 19.06%(57/299). The 4 false negative samples selected by these review rules did not come from the nephropathy department or the urology department.Microscopic results of RBC and WBC form these 4 samples ranged 3-8 cells/HP. Thus, these review rules could avoid the missed diagnosis of those patients with severe renal dysfunction.Conclusion The review rules established from this study for the UF-1000i urinalysis work station can effectively detect abnormal urine samples and improve the efficiency and the quality of urinalysis in routine clinical practice.

Objective To investigate the related factors on effects of uterine artery embolization (UAE) in the treatment of dysmenorrhea in patients with adenomyosis, and to construct and validate the efficacy prediction model. Methods A total of 127 cases of adenomyosis patients with symptoms of dysmenorrhea in Guangzhou No.1 People′s Hospital and Nanfang Hospital of Southern Medical University from June 1999 to December 2009 were reviewed. The evaluation standard was to improve the degree of dysmenorrhea, the related factors of efficacy were analysed. Combined with artificial neural network theory, the effect prediction model was constructed, and the effectiveness of the model was evaluated using receiver operating characteristic (ROC) curve, and the effectiveness of the cut-off point was calculated. The model was validated by 68 cases of patients with adenomyosis in the Nanfang Hospital from January 2010 to November 2014. Results (1) In 127 cases of dysmenorrhea patients, UAE treatment was effective in 98 cases, effective rate was 77.2% (98/127). (2) Age was an independent predictor of effective UAE treatment (HR=1.129, P=0.026);in the range of this study, the greater the age, the higher the UAE treatment efficiency. (3) The developing situation of ovary branches of uterine artery was an independent predictor of effective UAE treatment (HR=0.460, P=0.020), the efficiency of patients whose intraoperative bilateral uterine artery ovarian branch did not develop was 89.7%(35/39), the efficiency of patients whose unilateral uterine artery ovarian branch was developing was 84.1% (37/44) and the efficiency of patients whose bilateral uterine artery ovarian branch were developing was 59.1% (26/44). (4) Blood supply of adenomyosisis was an independent predictor of effective UAE treatment (HR=0.313, P=0.001). Type Ⅰ (bilateral predominated) patients, efficiency was 93.5%(43/46);typeⅡ(bilateral balanced) patients, efficiency was 78.0%(39/50);type Ⅲ (unilateral predominated) patients, efficiency was 51.6% (16/31). (5) UAE for the treatment of adenomyosis efficacy of artificial neural network prediction model was constructed, the model′s area under the ROC curve was 0.808, the optimal cut-off point was 0.669 13. Actual verification of the model, sensitivity was 96.5%, specificity was 81.8%, positive predictive value was 96.5%and negative predictive value was 81.8%, the total accuracy was 94.1%. Conclusions (1) Age, the developing situation of ovary branches and blood supply of adenomyosis are the independent predictors of effective UAE treatment. (2) The artificial neural network prediction model is satisfied with the accuracy and the accuracy of prediction.

Objective To construct human cytomegalovirus(HCMV)Han?ΔUS12?BAC strain and to study the role of US12 in HCMV replica?tion in human embryonic lung fibroblasts(HELF). Methods Kanamycin?resistant gene was amplified with primers containing homology arms se?quence flanking US12 and then electroporated into E.coli DY380?Han?wt?BAC competent cells. Successfully recombinant Han?ΔUS12?BAC clones were identified by PCR,sequenced for confirmation Han?ΔUS12?BAC plasmids were electroporated into HELF to produce infectious viri?ons. Han?ΔUS12?BAC and Han?wt?BAC were inoculated onto HELF at the multiplicity of infection of 0.1 pfu/cell. The viral titer in the supernatant was measured by TCID50 assay and growth kinetics of the viruses in HELF was studied. Results Han?ΔUS12?BAC clone was successfully con?structed. Han?ΔUS12?BAC was reconstructed in HELF to generate infectious virions. Growth kinetics assay indicated that Han?ΔUS12?BAC and Han?wt?BAC showed no differences in their growth and dissemination in HELF. Conclusion US12 in HCMV clinical strain Han is dispensable for HCMV growth and dissemination in HELF.

Objective To design and use the disposable puncture set in order to control the catheter related infection and implement conveniently the ultrasound-guided PICC catheter .Methods The disposable puncture set for PICC catheter was designed according to the operating process of ultrasound -guided PICC catheter.Results All tools in the puncture set had been arranged as the operation process , and was easy to use, and shielded completely the external pollution , and met the requirements of central venous catheter cluster intervention strategies .No case of infection was occurred in the 2 868 cases.Conclusions The disposable puncture set for ultrasound-guided PICC catheter is convenient to use , and can save time , and is worthy of clinical promotion .

As a new type of functional wound dressing, conductive hydrogel, shows broad prospects of application in the field of wound repair due to its suitable electrical conductivity, good moisture retention, excellent biocompatibility, and biological effects such as mediating cell migration and proliferation, and promoting angiogenesis and collagen deposition. Combined with the clinical electrical stimulation therapy, the conductive hydrogel primarily showed curative effects of promoting granulation tissue formation, re-epithelialization, and wound healing, providing a new treatment idea for the repair of diabetic wounds. This review summarized the research advances of electronic conductive hydrogels and ionic conductive hydrogels in recent years based on different conductive mechanisms. Meanwhile, the applications of conductive hydrogel in the diabetic wound repair were specifically introduced, and the future development of conductive hydrogel wound dressing was prospected.