As a new method of modern teaching,the computer assisted instruction(CAI) is widely used in every field in modern education. In this paper,the author explores the existed problems and the necessary qualities of using CAI for college teachers in microbiology experimental teaching.

Objective To construct the multiple antigenic peptide (MAP) gene and E. coli ex-pression system, based on the out membrane protein OmpL1, LipL21 and LipL32 from Leptospira interro-gans, and better understanding of the immunological activity of the recombinant protein. Methods Using MI3KE display and Western blot, the advantage epitopes of OmpL1, LipL21 and LipL32 were identified and used to synthesize a new gene, then its prokaryotic expression system was constructed. The expression of re-combinant protein was determined by SDS-PAGE. The immunity activity of the recombinant protein was iden-tified by Western blot and ELISA. Results The synthetic gene was effectively expressed in E. coli and mainly presented in soluble form. The expression protein could react with the antileptospirosis antibodies in rabbit and human sera, which contained different serogroups. Conclusion The recombinant MAP gene of leptospires was successfully constructedand and expressed in E. coli. The recombinant protein had a good immune activity, and could cross-reacted with antileptospirosis antibodies from different serogroups.

Objective To compare different equations for estimated glomerular filtration rate (eGFR) in patients with chronic kidney disease (CKD).Methods Hospitalized patients with CKD from the nephrology department of the First Affiliated Hospital of Nanjing Medical University (Jiangsu Province Hospital) were recruited between December 2014 and May 2015.The calculations of eGFR and 24 h creatinine clearance rate (Ccr) were accomplished in three days after admission.The eGFRs were calculated separately using the 24 h creatinine clearance rate adjusted by the standard body surface area (Ccr_BSA),Cockcroft-Gault equation adjusted by the standard body surface area (eCcr_BSA),CKD-EPI creatinine equation (EPI_Cr),CKD-EPI cystatin C equation (EPI_CysC),CKDEPI creatinine-cystatin C equation (EPI_Cr_CysC),simplified MDRD (MDRD) and China MDRD equations.The EPI_Cr_CysC equation was used as the standard and the precision and accuracy of the other six equations were compared and analyzed.Results A total of 403 CKD participants were enrolled in the study,with 228 male patients and a mean age of (54.9± 18.4) years.The main primary diseases were chronic glomerulonephritis (43.7％) and diabetic nephropathy (13.2％).The median concentration of serum creatinine and cystatin C were 117.5 (69.7,242.4) μmol/L and 1.80 (1.13,3.31) mg/L,respectively.The median values of Ccr_BSA,eCcr_BSA,MDRD,China MDRD,EPI_Cr,EPI_CysC and EPI_Cr_CysC equations were 50.8 (21.1,96.2),51.9 (23.3,93.2),53.6 (23.0,97.4),52.2(22.4,94.1),53.2 (22.1,97.3),35.1 (15.4,67.0) and 49.1 (22.8,82.3) ml · min-1 · (1.73 m2)-1,respectively.There was well agreement among MDRD,China MDRD and EPI_Cr equations,while there were large differences between equations derived from CysC (EPI_Cr_CysC and EPI_CysC) and equations derived only from creatinine (EPI_Cr,MDRD,China MDRD,eCcr_BSA,Ccr_BSA equations).Compared with EPI_Cr_CysC equation (the reference equation),EPI_Cr equation showed the highest accuracy [percentage of other eGFR equation calculations that were ＞ 30％ of the reference equation calculations (1-P30),30.8％] while Ccr_BSA equation showed the lowest (1-P30,42.4％).EPI_CysC equation showed the highest precision [inter-quartile range (IQR) of the difference,11.7 ml·min-1 · (1.73 m2)-1] while Ccr_BSA equation showed the lowest [IQR of the difference,22.8 ml· min-1 ·(1.73 m2)-1].Conclusions The agreement among equations derived only from creatinine is better;while it exhibits some differences between equations with cystatin C and equations derived only from creatinine.The accuracy of EPI_Cr equation is second only to EPI_Cr_CysC equation and it is currently the most suitable eGFR equation for clinical popularization of renal glomerular function assessment.

Background and purpose: With the development of the means of preoperative diagnosis and treatment of breast cancer, ifne needle puncture no longer seems to be popular with the clinical use. The safety and effectiveness of fine needle aspiration as preoperative diagnosis of breast cancer are controversial. This study was aimed to investigate the application of fine needle aspiration (FNA) of breast cancer and association between FNA and recurrence and metastasis for 30 years in our hospital. Methods:A total number of 1 260 patients with breast cancer treated in Peking Union Medical College Hospital from 1975 to 2006 were reviewed in two groups for clinical characters, recurrence, and survival rate. The two groups were FNA and non-FNA. Results:The positive rate of FNA was 61.4%and the highest rate was inⅣstage (100%). StageⅡandⅢwere the major ones in FNA group (45.2%). The local recurrence (7.0%vs 6.4%), recurrence and metastasis (13.2%vs 14.0%), 5-year (82.0%vs 81.0%) and 10-year disease free survival rates (51.0%vs 64.0%) have no signiifcant difference between FNA and non-FNA groups. Conclusion:FNA is still one of the most important methods for preoperative investigation of breast cancer. Background and purpose: With the development of the means of preoperative diagnosis and treatment of breast cancer, ifne needle puncture no longer seems to be popular with the clinical use. The safety and effectiveness of fine needle aspiration as preoperative diagnosis of breast cancer are controversial. This study was aimed to investigate the application of fine needle aspiration (FNA) of breast cancer and association between FNA and recurrence and metastasis for 30 years in our hospital. Methods:A total number of 1 260 patients with breast cancer treated in Peking Union Medical College Hospital from 1975 to 2006 were reviewed in two groups for clinical characters, recurrence, and survival rate. The two groups were FNA and non-FNA. Results:The positive rate of FNA was 61.4%and the highest rate was inⅣstage (100%). StageⅡandⅢwere the major ones in FNA group (45.2%). The local recurrence (7.0%vs 6.4%), recurrence and metastasis (13.2%vs 14.0%), 5-year (82.0%vs 81.0%) and 10-year disease free survival rates (51.0%vs 64.0%) have no signiifcant difference between FNA and non-FNA groups. Conclusion:FNA is still one of the most important methods for preoperative investigation of breast cancer.

Breast cancer is the leading cause of malignancy-related mortality in women worldwide. The more accurate prediction of lymph node metastasis and evaluation of personalized prognosis of breast cancer patients could provide evidence and reference for individualized comprehensive treatment and clinical decision-making. Nomogram is statistical calculation model developed to generate individualized prediction of a certain clinical event through the factors associated with it. Currently breast cancer related nomogram models is most commonly used in the prediction of non-sentinel lymph node status in patients with sentinel lymph node-positive breast cancer, sentinel lymph node metastasis in clinical node-negative breast cancer and prognosis evaluation of breast cancer. This article reviewed the recent advances in breast cancer related nomograms according to the above mentioned three aspects, and evaluated respectively the predictive factors, accuracy, characteristics and clinical application potential.

Objective:To analyze the distribution, clinico-pathologic features, and survival status of different subtypes in axillary lymph node-negative invasive breast cancer patients. Methods:In this study, data of 183 patients were included and retrospectively ana-lyzed in terms of age distribution, clinico-pathologic features, disease-free survival (DFS), and overall survival based on different sub-types (luminal, basal-like, and HER-2 over-expression). Results:No significant differences in age, tumor size, and TNM stage was ob-served among different subtypes. The relapse rates of luminal, basal-like, and HER-2 over-expression subtypes were 3.9% (4/102), 20.4% (10/49), and 6.3% (2/32), respectively (P=0.002). The death rates of luminal, basal-like, and HER-2 over-expression subtypes were 2.0%(2/102), 6.1%(3/49), and 3.1%(1/32), respectively (P>0.05). Kaplan-Meier analysis showed that the DFS of basal-like sub-type was much lower compared with that of the luminal and HER-2 over-expression subtypes (P=0.002). Cox analysis showed that the subtype was an independent prognostic indicator (P=0.001). Conclusion:In node-negative invasive breast cancer, no significant differ-ences in age distribution, tumor size, and TNM stage was observed among different subtypes. The basal-like subtype has the worst prog-nosis. Therefore, subtype is an important independent prognostic indicator.

Objective To clone mcp1, mcp2 and mop3 genes that encoding methyl-accepting chemotaxis proteins(MCP) of Campy/obacter jejuni for construction of their prokaryotic expression systems, and to establish chemotactic model in vitro of the microbe for determining chemotaxis-inducing substances and to understand relationship among MCPs and chemotactic inducers. Methods The segments of mep1, mcp2 and mcp3 genes were amplified by PCR and then sequenced after T-A cloning. Prokaryofic expression systems of the genes were subsequently constructed. SDS-PAGE pins Bin-Rod Gel Image Analyzer were used to examine the expression of target recombinant proteins rMCP1, rMCP2 and rMCP3, and Ni-NTA affinity chromatography was performed to purify the rMCPs. Rabbits were immunized with each of the three rMCPs to obtain antisera. Immunodiffusion assay was performed to measure the titers of antisera. IgG in each of the antisera were extracted by ammonium sulfate precipitation and DEAE-32 ion exchange chromatography, and IgG F(ab')2 were then prepared by pepsin enzymolysis and Sephadex G-100 chromatography. Chemotactic model in vitro of C. jejuni based on HAP( hard-agar plus) method was established to determine the chemo-taxis-inducing effect of eight candidate substances. Chemotaxis inhibition test based on IgG F(ab')2 bloc-king was applied to determine the function and diversity of MCPs. Results The segments with expected si-zes amplified from mcp1, racp2 and mcp3 genes were obtained by PCRs, and their nucleotide and putative amino acid sequences were completely same as the reported. The constructed prokaryotic systems could effi-ciently express rMCPs with the yields about 10% of the total bacterial proteins. Immunization with rMCP1, MCP2 and rMCP3 enables the rabbits to produce specific antibody. All the antisera had 1: 4 immunodiffusion titers. Both bovine bile and sodium deoxycholate(DOC) were able to induce ehemotactie movement of C. je-juni in a dosage-dependent manner ( P < 0.05 ). When MCP1 and MCP2 were blocked with their IgG F(ab')2, the ehemotaetic ability of C. jejurd were remarkably decreased (P <0. 05). However, MCP3 blocking did not affect the chemotaxis ( P > 0.05 ). Conclusion The C. jejuni MCPs are successfully ex-pressed in this study. Bovine bile and DOC are the inducers for chemotaxis of C. jejuni, and MCP1 and MCP2 are involved in the process of ehemotaxis iadueed by DOC.

Objective To screen the efficient antigenic epitopes in genus-specific envelope proteins OmpL1 and LipL21 of Leptospira interrogans for further development of multiple antigenic peptide (MAP)vaccine.Methods Based on bioinformatic technique,the combined epitopes of T and B lymphcytes in OmpL1 and LipL21 molecules were screened.Nucleotide fragments of each epitopes were amplified by PCR and then constructed their phage display systems.Using antisera against rOmpL1,rLipL21,L.interrogans serogroup Icterohaemorrhagiae strain Lai and leptospirosis patients' sera as the first antibodies.respectively,Western blot assays were performed to determine the immunoreaetivity and reactive ability of the epitopes with different antisera.Resuits Four combined epitopes of OmpL1 and two combined epitopes of LipL21 were selected out by the predicting procedure.All the amplified epitope fragments were accurately inserted into the region at N end of phage PⅢ protein and successfully expressed.All of the antisera could recognize each of the epitopes.Based on the results of Western blot,the two LipL21 epitopes at 97-112 and 176-184 showed similar strong hybridization signals with any of the antisera,and the hybridization signals of four OmpL1 epitopes with the three antisera were 173-191,87-98,297-320 and 59-78,from strong to weak.Conclusion The six combined epitopes in this study are efficiently antigenic.And the epitopes at positions 97-112 and 176-184 in LipL21 as well as the epitopes at position 87-98 and 173-191 in OmpL1 have a potential for developing leptospiral MAP vaccine.

Objective To investigate the effects of shRNA interference (RNAi) targeting the histone lysine specific demethylase 1 (LSD1) on the proliferation and apoptosis in acute leukemia (AL) cells. Methods LSD1 shRNA vectors were constructed and transduced into HL-60 and SHI-1 AL cell lines. Inhibitory efficiency of LSD1 was detected by real-time quantitative PCR (RT-qPCR) and Western blot respectively. Cell proliferation was assessed by methyl thiazolyl tetrazolium (MTT), and cell apoptosis was measured by flow cytometry. Results After interference of LSD1, the expression levels of LSD1 mRNA and protein in HL-60 and SHI-1 cells (mRNA: 0.242 ±0.023, 0.207 ±0.006; Protein: 0.256 ±0.015, 0.486 ±0.042) were decreased compared with blank control group without transfection process (mRNA: 1.021 ±0.178, 1.039 ±0.395; Protein:0.552 ±0.026, 0.754 ±0.060) and empty vector negative control group (shNC group) (mRNA: 0.935 ±0.136, 1.016±0.203;Protein: 0.500±0.026, 0.750±0.049) (P<0.05). The levels of their cell proliferation (absorbance value: 0.712±0.010, 0.549±0.007) were inhibited compared with blank control group (absorbance value:0.823±0.010, 0.625±0.005) and shNC group(absorbance value: 0.818±0.019, 0.621±0.003) (P< 0.05). While cell apoptosis rates were increased [(32.80 ±1.35) %, (23.49 ±1.40) %] compared with blank control group [(8.08 ±0.62) %, (7.28 ±1.17) %] and shNC group [(8.08 ±0.62) %, (7.28 ±1.17) %] (P< 0.05). Conclusions Lentivirus-mediated shRNA interferencing LSD1 can inhibit cells ' proliferation and promote apoptosis of HL-60 and SHI-1 AL cell lines, indicating that LSD1 may be a potential biological molecular marker and a new treatment target for AL.

Objective To evaluate the application of dynamic magnetic resonance imaging in the diagnosis of female stress urinary incontinence(SUI). Methods Dynamic magnetic resonance imaging(DMRI)were performed on 30 healthy female volunteers and 35 female SUI patients.DMRI of the pelvic floor at rest and OR maximal strain were performed by using sagittal T2-weighted fast gradient sequences.The distance of Urethra-vesical junction to the pubococcygeal line,the posterior vesicourethral angle and angle of inclination of the urethral axis were measured at rest and on maximal strain position.The t-value exact test were used to analyze the data. Results At rest the Urethravesical junction laid above pubococcygeal line on both control and SUI groups.Mean distance from the Urethra-vesical junction to pubococcygeal line at rest had no difference between the two groups.On straining,the mean Urethra-vesical junction descent distance in the SUI group(-0.9±1.1cm)was significantly higher than in control group(-0.14±0.3 cm),(P＜0.001).On straining,the mean angle of urethral inclination in the SUI group(65±37°)was significantly bigger than in control groups (17±21°),(P＜0.05).The posterior vesicourethral angle in the SUI groups(156±36°)was significantly bigger than in control groups(113±28°),(P＜0.05). Conclusion Dynamic magnetic resonance imaging is a non-invasive.easily applied method in the diagnosis of SUI.