Clinical skills training is a key factor for improvement of the quality of clinical teaching.Developing highly specific clinical skills training specification is an important step to improve the quality of gynecologic oncology clinical teaching.Based on its importance,a'four steps training strategy,known groups-building specification-train thinking-make a request is developed and used by Dept.of Obsterics and Gynecology,the 3rd Affiliated Hospital,Kunming Medical University.It guides students to achieve the basic theory,basic knowledge and basic skills through a comprehensive way,and it also encourages students with highly learning initiative of gynecologic oncology clinical skills.On the other side,in the clinical skills training will lead teachers to develop their thinking and enhance the enthusiasm of teaching.This method provides a new sight for gynecologic oncology clinical skills training in a new manner.

Minimal residual disease (MRD) is the cause of relapse in leukemia,and its dynamic monitoring has been a hotspot yet proven to be difficult.MRD in acute B-lineage lymphoblastic leukemia has its own characteristics,normal bone marrow produces a large number of B precursor cells,easily mistaken for leukemia cells,and there were no specific gene alterations that can be conveniently detected.This paper summarizes the antibody combinations in flow cytometry,timing for surveillance and polymerase chain reaction for gene detection,and provide a reference for B lymphocytic leukemia MRD monitoring.

Abstract Objective: To prepare high-purification and high-specific activity of recombinant human interleukin-6 (rhIL-6). Methods: The rhIL-6 was obstained from inclusion body expressed by IPTG-induced pT7.7hIL-6 expressed vector using extracting,denature and refolding techniques. The rhTL-6 was further purified by anionic-exchange chromatography. Activity of rhIL-6 was measured by 3H-TdR method. Results: After a single-step purification,the product purity reach 95% and it's specific activity was 3.0 x 10~8 U/mg. Conclusion:This scheme of puri-fication was an easy way requiring rhTL-6.

Objective To construct prokaryotic expression plasmid of human recombinant soluble TNF-related apoptosis inducing ligand(rsTRAIL),then to investigate the effects of rsTRAIL on apoptosis in non-small cell lung carcinoma(NSCLC).Methods The encoding sequence for rsTRAIL was amplified with RT-PCR and cloned into PQE30 vector to establish the prokaryotic expression system.The competent cells of host strain of M15 were transformed by the recombinant plasmid.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.rsTRAIL was added in the media of A549 and H460wt cells,then the viability was examined by MTT assay.Apoptosis of H460wt cells was observed under fluorescence microscopy.The apoptotic rate of tumor cells was examined by FACS.Results The cloned fragment of rsTRAIL was 100% consistent with that reported in GenBank.The expressed protein with molecular weight of 21 000 in SDS-PAGE as expected was obtained and recognized by a commercial McAb.The apoptotic rate of H460wt cells after treated with rsTRAIL for 24 h was 43.2%.Cislatin enhanced the effect of rsTRAIL on A549 cells.Conclusion The rsTRAIL is obtained after Ni affinity chromatograph.rsTRAIL has a strong cytotoxic activity against NSCLC and cisplatin may enhance the antitumor effect of rsTRAIL.

Objective To construct a recombinant expression vector of human interleukin-24(hIL-24) gene and express it in E.coli M15,and to evaluate the bioactivity of IL-24 fusion protein.Methods The human IL-24 cDNA fragment was amplified from plasmid by polymerase chain reaction(PCR),and sequenced.PQE/hIL-24 was constructed by gene rearrangement,then it was transformed into E.coli M15.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.The expressed recombinant hIL-24(rhIL-24) was identified by SDS-PAGE and Western blotting.Normal peripheral blood mononuclear cells(PBMCs) were cultivated with the expression protein for 48 and 72 h,the levels of IL-6,IFN-? and TNF-? of PBMCs stimulated with rhIL-24 were detected by ELISA.Results The recombinant prokaryotic expression vector PQE/IL-24 was constructed successfully and expressed stably in E.coli M15.At about 18 400 of molecular weight,there was an induced protein band.The levels of IFN-?,IL-6 and TNF-? in the group of cultivated with the expression protein were obviously higher than those in the groups without the expresson protein(P

Objective To assess the correlation between the pregnancy‐associated plasma protein A (PAPP‐A)level with gesta‐tional diabetes mellitus(GDM )in primiparous women .Methods The nonparametric test and Logistic regression analysis were used to retrospectively analyze 168 pregnant women of GDM and 273 normal pregnant women ,including age ,body weight ,PAPP‐A ,freeβ‐human chorionic gonadotropin(fβ‐hCG) .Results The age had significant difference between GDM group and control group(P<0 .01) ,the PAPP‐A concentration and corrected multiple of the median (MoM )of PAPP‐A in the GDM group were significantly lower than those in the control group(P<0 .01) .Conclusion Low level of PAPP‐A during early pregnant stage in primiparas is strongly associated with GDM and can be used as the risk factor of GDM .

ObjectiveTo observe the effects of chemotherapy on mitochondria ultrarnicrostructure and stereological characteristic of mucinous cystadenocarcinoma (MC) and serous cystadenocarcinoma (SC) and to investigate clinical significance and possible mechanism of chemotherapy on two cystadenocarcinoma.MethodsThe ultramicrostructure changes of mitochondrion were observed with electronic microscope and the stereological changes and its interrelation of volume density (Vv), surface density (Sv) and ratio of surface density(Rsv,μm-1)of mitochondria of the cells in ovarian carcinoma were studied and compared with the normal control. ResultsElectron microscope reveals that the mitochondria swelling and cristae disorganizing were seen in ovarian carcinoma.The Vv(0.072±0.059)and Sv(0.057±0.040)of MC mitochondria were decreased markedly(P =0.004 and P =0.029, respectively), while its Rsv (0.891 ±0.400) μm-1 increased obviously (P =0.015); but the Vv and Sv of mitochondria of the SC cells were increased markedly (0.267±0.12,P =0.001 and 0.122±0.057,P =0.003,respectively).After chemotherapy,cancer cells mitochondrion Sv elevated significantly (0.060±0.037, P = 0.053), showing a similar MC mitochondria of variability. Conclusion Therearedifferentcharacteristicsintheultramicrostructureandstereologicalcharacteristicinthe mitochondria of two types ovarian carcinoma. Effects of chemotherapy may be associated with the ameliorate of construction and function of ovary cells.

Objective To investigate the expression of the preferentially expressed antigen of melanoma (PRAME) gene in acute myeloid leukemia (AML),and to evaluate its applicability in monitoring minimal residual disease (MRD).Methods Bone marrow specimens were collected from 63 cases of de-novo AML,while 34 samples from 11 patients were tracked for 28 months.The level of PRAME mRNA was measured by real time RT-PCR.Results The PRAME gene expressed in 52.4 ％ (33/63) of de-novo patients,and the positive rate was highest in M3 than that in other subtypes of AML.The expression of PRAME became negative after treatment and increased in the following months before morphology relapse.Conclusion The PRAME gene is highly expressed in AML and could be a useful marker to monitor MRD.

Objective:To investigate the effects of different doses and infusion methods of recombinant mouse interleukin-18(rmIL-18) on the survival time and tumor diameter of the mice with hepatocellular carcinoma,and to elucidate the rational application of rmIL-18 in vivo.Methods:A total of 60 Babl/C mice were randomly divided into 5 μg rmIL-18 intraperitoneal injection group,0.5 μg rmIL-18 intraperitoneal injection group,0.5 μg rmIL-18 tumor injection group,cytotoxic T lymphocyte(CTL) intraperitoneal injection group,CTL tumor injection group and saline control group;there were 10 mice in each group.From the 10th day of inoculation,the mice in different rmIL-18 groups were injected with the corresponding doses and methods.The mice in different CTL groups were injected with tumor-specific CTL (1×106/mouse) by intraperitoneal and intratumoral injection.The mice in saline control group were injected with an equal volume (100 μL) of saline,the injections were performed 10 times.The diameters of mice were measured weekly and the survival time was recorded.Results:Compared with 5 μg rmIL-18 intraperitoneal injection group,0.5 μg rmIL-18 intraperitoneal injection group and saline control group,the tumor growth rate of the mice in 0.5 μg rmIL-18 tumor injection group was decreased (P<0.01)and the survival rate of the mice was increased (P<0.01);compared with 0.5 μg rmIL-18 intraperitoneal injection group,the tumor growth rate and the survival rate of the mice in CTL intraperitoneal injection group were decreased (P<0.01);compared with 0.5 μg rmIL-18 tumor injection group,the tumor growth rate and the survival rate of the mice in CTL tumor injection group were decreased (P<0.01).Conclusion:The best way for rmIL-18 anti-tumor effect is tumor injection and the effect has a dose-dependent manner.

Objective:To research rmIL-18 in vitro culture system CCs induce tumor-specific cytotoxic T lymphocytes CTL and anti-tumor effect in mice.Methods:Used Stem SepTM immune magnetic cells separation method to culture mouse spleen NK cells,T cells and DCs,established culture systems in vitro;used of different approaches,different doses rmIL-18 to immunize HCC tumor-bearing mice,researched the effect of rmIL-18 on tumor growth rate and survival time.Results:rmIL-18 could induce and promote tumor-specific CTL-mediated killing effects in vitro culture system;tumor-specific CTL could significantly inhibit tumor growth(P<0.01) of and prolong the survival time of liver cancer tumor-bearing mice(P<0.01),and the effect was increased with rmIL-18 concentration increased(P<0.01),and intratumoral injection was superior to intraperitoneal injection(P<0.01).Conclusion:rmIL-18 can induce tumor-specific CTL in vitro and play a role in anti-liver cancer in mice.