Objective To explore the clinical significance and the impact on prognosis of blood micrometastasis in the patients with pN0 esophageal squamous cell carcinoma. Methods Total RNA was extracted with TRIzol and mRNA was transcribed reversely into cDNA. RT-PCR was used to detect MMP-7 mRNA and hTERT mRNA in blood. △△Ct sample values were calculated with post-operative follow-up of 3 month, 6 month, 12 month. Results Statistical results suggested that blood micrometastasis was related to differentiation grade and pTNM staging (P=0.000, P=0.000 respectively), but not to age, sex, length of turnout (P0.05). Follow-up results suggested that the degree of invasion and tumor metastasis (recurrence) was no correlation; follow-up to 6 month and 12 month, tumor metastasis (recurrence) was associated with blood micrometastasis, and follow-up to 12 month, compared with the tumor metastasis (recurrence) probability of blood micrometastasis-positive patients and negative patients, the former was as 6.44 times as the latter. (OR=6.440, 95 % CI 1.547-26.822). Conclusion Blood micrometastasis testing is of great significance to early diagnosis and prognosis judgment in pN0 esophageal squamous cell carcinoma patients.

ObjectiveTo summarize the nursing measures for patients with nasal facial soft tissue defect treated with transplantation of free foot skin flap.MethodThe clinical data of 13 patients with transplantation were reviewed to summarize the nursing measures.ResultAll skin flaps of the 13 patients survived.ConclusionEnough preoperative preparation,active psychological nursing and careful postoperative observation are critical for the success of the operation.

OBJECTIVE:To establish the quality standard for Prostatitis capsules. METHODS:TLC method was performed to qualitatively identify Pseudostellaria heterophylla,Acortw tatarinowii,Sargentodoxa cuneata,Rubia cordifolia and Vaccaria segeta-lis in the preparation. HPLC method was adopted to determine the content of leonurine in the preparation. The determination was performed on Agilent Eclipse XDB-C18 column with mobile phase consisted of methanol-0.025 mol/L potassium dihydrogen phos-phate(pH adjusted to 2.5)(24 : 76,V/V)with phosphoric acidat the flow rate of 1.0 mL/min. The detection wavelength was set at 277 nm,and column temperature was 30 ℃. The sample size was 10 μL. RESULTS:The TLC spots of P. heterophylla,A. tatari-nowii,S. cuneate,R. cordifolia,and V. segetalis were clear and well-separated without interference from negative control. The lin-ear range of leonurine were 4.05-81.00 μg/mL(r=0.9999). RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recovery was 98.47%-103.83%(RSD=2.04%,n=9). CONCLUSIONS:Established standard can be used for quali-ty control of Prostatitis capsules.

Objective To investigate the effect of folic acid combined with Helicobacter pylori (Hp) eradication therapy on chronic atrophic gastritis.Methods From December 2009 to March 2011 at the First Affiliated Hospital of Nanjing Medical University,184 patients with endoscopic and pathological diagnosis of chronic atrophic gastritis (90 Hp positive and 94 Hp negative) were selected.Hp positive patients were divided into group A and group B.Forty-three patients in group A were treated with standard triple Hp eradication therapy and follow by folic acid therapy for three months.Forty-seven patients in group B and Hp negative patients received three months of folic acid therapy.The clinical symptoms of each group were scored before treatment,one month after folie acid therapy and three months after folic acid therapy and analyzed by t test. Patients of each group received gastroscopy before treatment and three months after medicine withdrawal. Endoscopic scores,pathological scores and t test were recorded.The serum levels of pepsinogen ( Ⅰ,Ⅱ ) and gastrin 17 in venous blood of 55 Hp negative patients were detected by enzyme-linked immunosorbent assay (ELISA) method before treatment and three months after medicine withdrawal.Results Compared with three months therapy (1.15 ± 0.03),after one month folic acid therapy (1.55 ± 0.04) was statistically significant in clinical symptoms score of all patients (t =8.18,P＜0.01).By the end of therapy,clinical symptom score of group A (1.06 ± 0.04) was lower than that of group B (1.56 ±0.08),and the difference was significant (t=6.00,P＜0.01).There was significant difference in endoscopic scores of all patients between before treatment (1.57±0.95) and after treatment (1.00±0.76,t=11.12,P＜0.01).The differences in each pathological score of all patients (inflammatory scoring,active scoring,atrophy scoring,intestinal metaplasia scoring,atypical hyperplasia degree scoring) were significant between before treatment and after treatment (t=5.51,6.90,7.53,6.34,2.90,respectively,all P＜0.01).The serum level of pepsinogen Ⅰ before treatment of 55 Hp negative patients [(1.03±0.19) nmol/L] was lower than that after treatment [(2.24±0.33) nmol/L],and the difference was statistically significant (t=3.19,P＜0.01).After treatment the level of gastrin 17 [(0.86±0.05) nmol/L] was higher than that before treatment [(0.47±0.05) nmol/L],and the difference was statistically significant ( t =5.33,P＜ 0.01 ).Conclusion Folic acid in combination with Hp eradication therapy can be favorable to atrophic gastritis,which may promote the secretion of pepsinogen and gastrin.

According to the constructivism approach, instructors have to adapt to the role of fa-cilitators but not teachers. Whereas a teacher gives a didactic lecture that covers the subject matter , a fa-cilitator helps the learner to get to his or her own understanding of the content. In the former scenario the learner plays a passive role and in the latter scenario the learner plays an active role in the learning pro-cess. Under the guidance of this theory, a multi-dimension teaching model based on classroom teaching, network platform and innovate experiments has been established in the course of basis of clinical labora-tory. It has been found that this model is conducive to raising students' interests in learning and to culti-vating student's comprehensive quality.

It is for estimating the laboratory statistics from the view of clinical medicine objectively,completely and appropriately that we establish ＜ Laboratory Sciences and Clinical Medicine ＞ which fits the requirement for the modern medical laboratory specialists. In this selective course, we use problem-based learning ( PBL ) teaching method which is different from the traditional one, stimulating students to participate in the study actively and passionately,training them to think in a scientific way and to analyse and solve the problem in a rational way. As a result, it works well which makes us believe it is worth popularizing.

Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .

Objective To investigate the epidemiological characteristics of dengue fever in Shenz-hen city in 2014 and to analyze the evolutional characteristics of the epidemic dengue virus(DENV)strains in order to provide scientific guidelines for the prevention and control of dengue fever. Methods Descrip-tive epidemiological analysis was used to analyze the prevalence of dengue fever in Shenzhen city in 2014. Immunochromatography and real-time PCR were performed to detect the specific antibodies(IgM and IgG) and DENV nucleic acids in serum samples collected from suspected cases of dengue fever. Serum samples collected from the patients at early stage of dengue fever were used to infect the C6 / 36 cell line for further isolation of DENV strains. The types of isolated DENV strains were determined by using real-time PCR. E genes of the isolated DENV strains were amplified by RT-PCR and then sequenced. DNAStar and Clustslx (1. 83)softwares were used to analyze the homology between DENV strains isolated in Shenzhen and other areas. A phylogenetic tree based on the sequences of E genes of Shenzhen strains and other sequences of DENV reference strains downloaded from GenBank was constructed for further analysis. Results A total of 454 cases of dengue fever were reported in Shenzhen in 2014 with a male to female ratio of 1. 43 ∶ 1. Local patients accounted for 76. 21% and the rest 23. 79% were imported cases mainly from Southeast Asian and surrounding cities. There were 441 cases reported from September to November,accounting for 97. 14% of all reported cases. Most of the infected subjects were aged 20 to 50,accounting for 76. 73% . Of the 270 samples positive for DENV nucleic acids,strains of DENV-1,DENV-2,DENV-3 and DENV-4 accounted for 87. 41% ,8. 89% ,0. 37% and 2. 22% ,respectively. The phylogenetic tree analysis revealed that the DENV-1 strains belonged to two genotypes,which were genotypeⅠ and genotype Ⅴ. The DENV strains of genotypeⅠ were highly similar to the epidemic strain isolated in Shenzhen in 2010 and the genotype Ⅴstrains were first reported in Shenzhen. The homology analysis of the nucleotides of E genes showed that mi-nor differences in the nucleotide sequences were found between DENV-2 strains. All of the DENV-2 strains belonged to the genotype Ⅳ as indicated by the phylogenic tree. Conclusion There were 454 cases of den-gue fever(including both local and imported cases)reported in Shenzhen city in 2014,reaching an all-time high. DENV-1 was the predominant pathogen in combination with an increased infection of DENV-2. This study indicated that the prevalent DENV strains might be imported from Southeast countries and neighboring cities. Further researches should be conducted to analyze whether dengue fever is endemic in Shenzhen City.

The oncogenic isoform of the p63 protein, delta NP63, plays an important role in the pathogenesis of many epithelial carcinomas, and emerging evidences suggest that delta NP63 is a promising drug target. However, the functions of delta NP63 in transitional cell carcinoma of bladder (TCCB) are poorly defined. In this study, a delta NP63 shRNA expression vector was transfected into TCCB cell line 5637 and cell cycling, cell proliferation and protein expression were assessed by flow cytometry and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dimethyl tetrazolium bromide (MTT) assay, and immunohistochemistry, respectively. The delta NP63 shRNA expression vector was also injected into 5637 cell xenograft tumors in nude mice, and tumor size was measured, tumor tissue morphology was assessed by immunohistopathology and transmission electron microscopy. In the in vitro study, delta NP63 shRNA transfection caused successful delta NP63 gene silencing and resulted in significant arrest of cell cycling and cellular proliferation (p<0.05) as well as cyclin D1 expression. In the nude mouse xenograft model, delta NP63 shRNA greatly inhibited tumor growth, induced tumor cell apoptosis (p<0.05) and resulted in cyclin D1 downregulation. Our data suggest that delta NP63 may play an oncogenic role in TCCB progression through promoting cell survival and proliferation. Intratumoral administration of delta NP63-specific shRNA suppressed tumor delta NP63 expression and cellular proliferation while promoted tumor cellular apoptosis, and therefore inhibited tumor growth and improved survival of xenograft-bearing mice, which was not accompanied by significant signs of systemic toxicity.
Animals ; Carcinoma, Transitional Cell/*genetics/metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1/biosynthesis ; Disease Progression ; Female ; Humans ; Mice ; Mice, Nude ; Microscopy, Electron, Transmission ; Models, Biological ; Neoplasm Transplantation ; Trans-Activators/*biosynthesis/*physiology ; Tumor Suppressor Proteins/*biosynthesis/*physiology ; Urinary Bladder Neoplasms/*genetics/metabolism

OBJECTIVE:To optimize the extraction technology of caffeic acid in Laggera alata,and establish a method for its content determination. METHODS:The caffeic acid in L. alata was extracted by reflux extraction. Using extraction content as inves-tigation index,orthogonal test was used to investigate the effects of ethanol volume fraction,material-liquid ratio and extraction time on caffeic acid,and the extraction technology conditions were optimized. HPLC was adopted to determine the content of caffe-ic acid in 10 batches of L. alata from different areas,using caffeic acid as reference substance,at wavelength of 320 nm. RE-SULTS:The optimized extraction technology conditions were as follows as ethanol volume fraction of 10%,material-liquid ratio of 1 : 40 and extraction time of 3 h. Under the condition,verification test for caffeic acid was carried out,and the average content of caffeic acid in L. alata was 0.5211 mg/g(RSD=1.18%,n=3). The content of caffeic acid in 10 batches of L. alata from dif-ferent areas ranged in 0.3752-0.7766 mg/g,and the content showed great differences. CONCLUSIONS:The content of caffeic ac-id in L. alata is related to area and harvest season. The caffeic acid extration by optimized technology shows good reproducibility;and the established method for content determination is stable and feasible.