The construction of key laboratory in colleges is the important warranty of the upgrading of school's comprehensive research competence.In this text,we have discussed how to strengthen the building of key laboratory and facilitate the overall development of research system in university,and considered the essentiality of construction and the reason of development.

This paper summarizes that to strengthen the construction of national key disciplines Clinical Laboratory Diagnostic and build professional post-graduate education platform for innovation,the exploration and practice of reform have been done in Chongqing Medical University,in the teaching ranks and exercise disciplinary research direction to enhance the level of scientific research and laboratory construction.

For the reform and development of clinical laboratory education, based on the previous course reform, Laboratory Medicine College of Chongqing Medical University has put the leading teachers'responsibility system of laboratory medicine course into practice. In the recent 3 years, the course is better organized.The students are more interested in the course and they communicate more with teachers than ever before. The effect of the course is obvious. The leading teachers' responsibility system in laboratory medicine course should be promoted.

Objective To construct the RNA interference eukaryotic expression vector targeting human centromere protein-I (CENP-I) and to observe its effect on the growth of human embryo kidney 293 cells (HEK 293). Methods The expression vectors of pGenesil-1/CENP-I-siRNA-1. pGenesil-1/CENP-I-siRNA-2 and pGenesil-1/CENP-I-siRNA-3 were constructed by gene recombination and then were transfected into the HEK293 cells by liposome. The expressions of CENP-I at the protein and mRNA levels were detected by Western blotting and fluorescence quality PCR (FQ-PCR). The effective vector and the best transfection time were selected. The growth and the cell cycle of the transfected cells were assessed by MTT assay and flow cytometry. Giemsa was used to stain the transfected cells to calculate the mitotic index. Results Sequence-specific siRNAs targeting CENP-I significantly down-regulated the expression of CENP-I in HEK293 cells. The recombinant plasmid of pGenesil-1/CENP-I-siRNA-3 was the effective vector. After transfecting for 72 h the best inhibited efficiency was achieved. In CENP-I-siRNA transfected cells,the rate of cell growth was decreased markedly. Cells at G 2/M phase and the mitotic index were increased conspicuous compared with the cells transfected with the blank vector or untransfected. Conclusion Down-regulation of CENP-I in HEK293 cells by sequence specific siRNA delays the cell growth and postpones the cell division.

Objective To explore the mechanism of hBUB1 gene in the developing of spontaneous abortion embryos with numerical chromosomal abnormalitv.Methods Quantitative real-time RT-PCR and Western blot were used to determine the mRNA and protein level of hsMAD2 gene both in spontaneous abortion embryos with numerical chromosomal abnormality(experimental group)and with numerical chromosomal normality(control group).Recombinant shRNA plasmids targeting hBUB1 gene was constructed to inhibit the expression of endogenous hBUB1 genes in embryonic cells.Interference efficiency was demonstrated by fluorescent quantitative PCR and Western blot.The inhibitory rate of cell proliferation was measured by MTT assay and cells-cycle was assessed by flow cytometry.Resuns Western blot analysis showed that protein level of hBUB1 in the experimental group was decreased signifieandv(the rate of positivity and strong positivity were 8%and 93.5%,respectively,P＜0.05)compared with the control group.The expression of hBUB1 gene in embryonic cells was significantly and specially inhibited by shRNA plasmids (the mRNA level before and after treatment witll RNAi were 0.196±0.067 and 0.042±0.006,respectively,P＜0.05).The inhibitory rate of cell proliferation was increased to 62%from 4%at 48 h after transfection.The rate of G2/M phase cells was decreased after transfection with efficient shRNA(control group:40.2%and 41.3%,test group:21.3%).Conclusions Down-regulation of hBUB1 gene leads to the inhibition of cell proliferation and the arrest of cell-cycle.It also probably plays an important role in the development of spontaneous abortion embryos with numerical chromosomal abnormality.The clinical relevance warrant further investigation.

Objective To investigate the expression of human spindle mitosis arrest deficiency gene (hsMAD2)in spontaneous abortion embryos and the relationship between low expression of hsMAD2 and numerical chromosomal aberration. Methods Spontaneous abortion embryo tissues were collected,including 23 cases of once spontaneous abortion tissue and 10 cases of twice or more spontaneous abortion tissue and induced abortion embryos(35 cases)from the Department of Gynaecology and Obstetrics of the Affilisted Hospitals of Chongqing University of Medical Science during the period of March 2006 to March 2007.FQ-PCR and western blot were used to evaluate the endogenous expression level of hsMAD2 mRNA and hsMAD2 protein;primary culturing of cells from the induced abortion embryos was conducted and 5 embryonic cells were selected by chromosomes karyotype analysis.Recombinant shRNA plasmids targeting hsMAD2 gene were constructed to inhibit the expression of endogenous hsMAIY2 genes in embryonic cells which have normal karyotypes;the groups were defined as the first experimental group(transfeeted with pshRNA-hsMAD2-1),the second experimental group(transfected with pshRNA-hsMAD2-2),the third experimental group(transfected with pshRNA-hsMAD2-3),the first control group(transfected with nothing),the second control group(transfected with pTZU6+1)and the independent group(transfected with pshRNA-N1).Interference efficiency was demonstrated by FQ-PCR and western blot:cell prolireration was meagured by methyl thiazolyl tetrazolium(MTT)assay;cell-cycle was assessed by flow cytometry (FCM):the chromosome numbers were calculated to analyze the variation of chromosomes.Results(1) The mRNA levels of hsMAD2 in the once spontaneous abortion tissue,twice or more spontaneous abortion tissue and indueed abortion tissue were 0.00879±0.00035.0.00901±0.00033 and 0.00941±0.00026 respectively,and there Wag no significant ditierence(P＞0.05)compared with each other;however,the protein levels of hsMAD2 in three groups were 0.2791±0.0311.0.0431±0.0020 and 0.5790±0.0331 respectively,and there were significant difierences(P＜0.05)compared with each other.(2)Recombinant shRNA plagmids could significantly and specifically inhibit hsMAD2 gene expression in embryonic cells.Compared with the first control group(4%)and the second control group(3%),the recombinant shRNA could inhibit embryonic cell proliferation to 54% at 48 h after transfection(P＜0.05):compared with the first control group(8.2%)and the second control group(8.0%),the ratios of G2/M phase cells in the experimental group(17.9%)was significantly increased(P＜0.05);compared with the first control group (4.8%),the ratios of abnormal chromosomes in the experimental group was increased to 30.0%(P＜0.05).Conclusions Down-expression of hsMAD2 gene may be one of the mechanisms inducing numerical chromosome aberration,abnormal embryo development and the occurrence of spontaneous abortion.

Objective To study the diagnostic value of combined detection of serum CEA,CA199,CA125,AFP and fecal occult blood(FOB) in Dukes B stage colon carcinoma.Methods The above indicators in 56 patients with colon carcinoma (colon carcinoma group),35 patients with colonic polyp benign disease and 41 healthy individuals undergoing physical examination were measured.Then the sensitivity,and specificity of single index detection and their combined detection were analyzed.Results The levels and positive rates of CEA,CA199,CA125 and FOB in the colon cancer group were significantly higher than those in the colonic polyp group and healthy control group (P＜0.05),but the AFP level and positive rate had no statistical difference compared with the colonic polyp group.Serum CEA,CA125,CA199 and FOB were selected into the Logistic regression equation.The corresponding areas under the curve (AUC) were 0.745,0.886,0.792 and 0.864 respectively;the positive detection rates were 48.2 ％,35.7 ％,39.3 ％ and 78.6 ％ respectively.The combined detection could increased the specificity,sensitivity and AUC to 87.5 ％,92.1％ and 0.957,which were more superior to that in the single index detection.Conclusion The combined detection of serum CEA,CA199,CA 125 and FOB can provide the powerful basis for early diagnosing colonic cancer.

Guided by the demands of society,we propose a new professional training objectives of medical laboratory technology,which is to cultivate comprehensive talents covering the entire industrial chain of medical laboratory technology with profound foundation,broad caliber,high quality and outstanding ability.According to the training objective,we build up 4321 talents training system and try to carry out preliminary practice and exploration on talent cultivation model from the following aspects,such as the construction of curriculum system,the reform of the teaching contents and methods,training of students' professional skills and entrepreneurial innovation spirit.

OBJECTIVETo investigate the effect of high-intensity focused ultrasound (HIFU) on tumor metastasis in mouse model bearing melanoma xenograft.

METHODSMice bearing murine melanoma B16-F10 cell xenograft were randomized for sham-HIFU or HIFU exposure when the tumors grew to a maximum diameter of 7-10 mm, and the tumor size was measured every 3 days. The cumulative survival rate of the mice and tumor metastasis rate were calculated, and the circulating melanoma cells were detected using qRT-PCR. At 14 days after HIFU treatment, B16-F10 cells were retransplanted via the tail vein and the pulmonary metastatic nodules were counted.

RESULTSThe median survival time of the mice was 19.00 days (95% CI 17.14-20.86 days) in the sham group and 26.00 days (95%CI 24.76-27.25 days) in HIFU group. The cumulative survival rate in the HIFU group was significantly higher than that in sham-HIFU group (P<0.01), and the tumor size was significantly smaller in HIFU group at 20, 23, and 26 days after HIFU treatment (P<0.05). Compared with the sham-HIFU group, HIFU group had significantly lower levels of MAGE-A3, MART1 and PAX3 at 7 days after HIFU (P<0.05) with still lower MAGE-A3 level at 14 days (P<0.05). HIFU group showed a significantly smaller number of pulmonary metastatic nodules following tumor cell retransplantation than in sham-HIFU group (P<0.01) with a metastasis inhibition rate of 42.4%.

CONCLUSIONHIFU treatment can inhibit tumor metastasis in melanoma-bearing mice possibly by reducing tumor cell detachment from the primary tumor site and suppressing colonization of the circulating melanoma cells.

Objective:To study the outcomes and post-discharge follow-up of neonatal tracheotomy in neonatal intensive care unit(NICU).Methods:This study included patients who were admitted to NICU in Beijing Children′s Hospital from January, 2016 to August, 2021, and less than 28 days or 44 weeks(corrected age)on admission, and required tracheotomy.The patients were divided into tracheotomy group and the non-tracheotomy group (the parents signed to refuse the tracheotomy) according to whether perform tracheotomy.Demographic data, general hospitalization information, diagnosis, indications for tracheotomy, follow-up outcomes at 3/6/12 months of age after discharge of patients were collected and analyzed.Results:Totally 26 patients were included in this study, 14 cases in tracheotomy group and 12 cases in non-tracheotomy group.The average gestational age was(37.7±3.80)weeks and(38.99±1.83)weeks, and birth weight was(2 823.57±948.89)g and (3 320.83±378.76)g, respectively.There were no significant differences in sex, gestational age, birth weight, age on admission, weight on admission, age at diagnosis, ratio of endotracheal intubation for respiratory support on admission between two groups( P>0.05). The commonest indications of tracheotomy group were bilateral vocal cord paralysis(50.0%) and congenital anomaly/defect of throat/larynx(21.4%), and the commonest indications of non-tracheotomy group were bilateral vocal cord paralysis(50.0%) and vocal cord/subglottic mass(25.0%), and there was no significant difference between two groups( P>0.05). The rate of discharge-against-medical order of tracheotomy and non-tracheotomy group was 7.14% and 66.67%( P=0.003), respectively.The total follow-up rate of tracheotomy and non-tracheotomy group was 88.9% and 38.9%, while the follow-up rates at 3 months, 6 months, and 12 months were 100.0% vs. 50.0%, 83.3% vs. 41.7%, and 81.8% vs. 25.0%, respectively, whose differences were statistically significant(all P<0.05). In the 14 cases of tracheotomy group, 3 cases died, 4 cases successfully removed the tracheal cannula, 5 cases did not remove the tracheal cannula, and 2 cases were lost. Conclusion:Bilateral vocal cord paralysis is the commonest indication of neonatal tracheotomy.Parents′ compliance in the tracheotomy group is significantly higher than that in non-tracheotomy group.To give caring skill training for parents of neonates with tracheotomy before discharge is beneficial for improving the overall prognosis of children.