Objective To investigate the effect of cervical knife conization (CKC) or loop electrical excision procedure (LEEP)on the outcome of subsequent pregnancies and mode of deliveries. Methods A retrospective case-control study including 228 women after treatment with LEEP or CKC for cervical intraepithelial neoplasia (CIN) Ⅱ -Ⅲ who gave birth in the First Affiliated Hospital of Sun Yat-sen University and He-xian Memorial Hospital of Pangyu from January 2004 to January 2010 was performed.Patients (n = 228) without cervical surgical history were randomly extracted from the respective hospitals birth registries as controls and were matched by age, gestation,parity and income.The information including gestational age, premature rupture of membranes (PROM), type of deliveries and birth weight of the two groups were collected.Results The gestational age of women treated with conization was (268.3±26.2) d, longer than that of the women without surgery (279.4±25.3) d (t=4.60, P＜0.01). The incidence of preterm birth was 18.0％(41/228) and 4.4％ (10/228) (x2 = 21.22, P＜ 0. 05). The incidence of PROM was higher in conizationgroup (10.1％, 23/228) than that (1.3％, 3/228) in control group (x2=16.32, P＜0. 05). Risk for PROM was almost eight fold (OR=8. 42, 95％CI: 2.49-28.44) higher in conization group. Cesarean section rate was higher in conization group (69.3 ％ ) than in control group (39.0 ％ )(x2=42.06, P＜0. 01). The gestational age of women treated with LEEP was longer than those treated with CKC[(269.8±24.6) d vs (260.2± 26.5) d, t= 4. 01, P＜0.01]. The incidence of preterm birth was 13. 1％ (22/168) and 31.6％ (19/60) (x2 = 10. 34, P＜0. 05). The mean birth weight of women with LEEP was heavier than that with CKC[(3358.5 ±812.2) g vs (3295.9 ±832.6) g, t=3.08, P＜0. 01]. The incidence of PROM (7.1％, 12/168) of woman with CKC was higher than that (1.3％, 11/60) of women with LEEP (x2 =6.10, P＜0.05). Conclusions Conization might increase the incidence of preterm delivery and preterm PROM. LEEP showed less adverse effect onthe outcome of subsequentpregnanciesthan CKC,and waspreferredfor primigravida, and the risk of treatment should be informed in advance.

AIM: To study the mRNA and protein expression of Kang ai1 (KAI1) tumor suppressor gene and to determine the relationship between KAI1 and invasiveness and metastasis of cervical cancer. METHODS: The expression of KAI1 metastasis suppressor was detected by immunohistochemistry in paraffin slides and by real-time quantitative polymerase chain reaction (RT-PCR) in fresh tissue. The samples included 20 cases of normal cervical tissues, 20 cases of cervical intraepithelial neoplasia (CIN) and 40 cases of cervical carcinoma. The results of the gene expression combined with the pathological and clinical data were also analyzed. RESULTS: The expression of KAI1 protein and mRNA was related to the tissue differentiation of cervix. The positive rates of KAI1 expression were the highest in the normal cervical tissue, the middle in CIN and the lowest in cervical carcinoma with significant difference among three groups (P<0.01). The expression of KAI1 protein was not related with the grade of CIN (P>0.05). However, both mRNA and protein expression of KAI1 were related to the differentiation and the clinical stages of cervical cancer (P<0.01) and also related to the metastasis of the cancer. The positive rates between the non-lymphatic metastasis and lymphatic metastasis (P<0.05) were significant different. Cox regression and logistic regression showed that the tissue differentiation, clinical stages, lymphatic metastasis and expression of KAI1 were all related factors with recurrence and prognosis of cervical cancer. CONCLUSION: The down-regulation of KAI1 tumor suppressor gene at both mRNA and protein levels is related to the differentiation, clinical stages and metastasis of cervical cancer, indicating that the expression of KAI1 is a prognostic factor for cervical cancer.

Objective To construct the murine allogeneic acute GVHD model.Methods C57BL/6 (H-2b) mice were used as the donors and Balb/c (H-2d) mice as the recipients in allogeneic bone marrow transplantation (BMT).Groups were set as total body radiation (TBI) control group (n =4),GVHD group (n =10),simple BM transplantation group (n =10) and normal control group (n =4).For TBI control group,mice were subjected to TBI but did not receive BMT after radiation.For GVHD group,5 days before TBI,gentamycin (320 mg/L) and erythromycin (250 mg/L) were added into the drinking water,and on the day of transplantation,mice received one total dose of 8.0 Gy 60Coγ TBI,and within 5 h,2 × 106 C57BL/6 BM cells and 1 × 107 C57BL/6 spleen cells were transfused per mouse via the tail vein.For simple BMT group,the pretreatment was the same as GVHD group,and mice received only 2 × 106 C57BL/6 BM cells per mouse via the tail vein.The mental status,activity,posture,fur,weight,and stool were observed after transplantation.Survival time of each mouse was recorded,survival rate was calculated,and survival curve was drawn.Pathological examination was done for the liver,skin,small intestine and BM on the brink of death.Results The median survival time (MST) in TBI control group,GVHD group and BMT group was (9.0 ± 0.7),(32.0 ± 3.2) and ( 17.5 ± 1.6) days respectively,and there was significant difference between every two groups (P ＜ 0.01 ).Pathological examination in TBI control group showedhematopoiesis exhaustion.GVHD group showed acute GVHD symptoms 10-13 days after allo-BMT,and the pathological changes of the skin,liver and small intestine corresponded to those of Ⅰ to Ⅱ degree of GVHD.Simple BMT group also showed acute GVHD symptoms 10-13 days after alloBMT,but their GVHD manifestation and histological changes were less serious and only 0 to Ⅰ degree of GVHD could be seen.ConclusionStable acute GVHD model can be constructed by transfusion of allogeneic BM cells and spleen cells into Balb/c mice after lethal TBI.

[Objective] To investigate the relationship of different types of gestational diabetes mellitus (GDM) and thyroid function.[Methods] A Total of 3846 cases,which received prenatal examination,delivered in the Eastern Hospital of the First Affiliated Hospital,Sun Yat-sen University and performed a 75 g oral glucose tolerance test (75 g 0GTT) at 24-28 gestational weeks,from Jan 1st,2014 to Dec 31st,2015,were divided into 2 groups.Normal blood glucose group:the result of OGTT (fasting plasma glucose,1 hour glucose and 2 hour glucose) was normal;Gestational diabetes mellitus group (GDM group):the result of 0GTT was abnormal.GDM group were divided into Ⅰ,Ⅱ,and lⅢ.GDM Ⅰ defined as one abnormal blood glucose of result.GDM Ⅱ:two abnormal blood glucose.GDM Ⅲ:three abnormal blood glucose.1868 cases of healthy pregnant women were reselected as the control group.TSH,FT4 and TPO Ab were detected in two groups.Analysis of Variance,Mann-Whitney U test,Kruskal Wallis rank test or Fisher's test was used for statistical analysis.[Result] There were statistically significant difference in TSH,FT4 between GDM subgroup and control group (P =0.012,P =0.002).TSH median trend to increase in GDM Ⅱ,and FT4 median trend to decrease in GDM Ⅱ.The Prevalence of hypothyroidism in GDM Ⅱ and GDM Ⅲ were higher than those in control group.[Conclusion] The GDM group with two or three abnormal blood glucose had a higher incidence thyroid gland dysfunction,especial with subclinical hypothyroidism.We should fully test the thyroid function,treat diabetes as early as possible and improve the pregnancy outcome as we could.

[Objective] To investigate associations between the functional polymorphisms of signal transducer and activator of transcription 4 (STAT4) gene and unexplained recurrent spontaneous abortion (URSA) and between STAT4 protein expression and the genotypes of rs 10181656 locus.[Methods] PCR-restriction fragment length polymorphism was used to genotype rs 1 0181656 locus polymorphism in 332 URSA cases and 260 normal controls,in 86 URSA cases and 77 normal controls of which immunohistochemical technique was used to detect STAT4 protein expression.[Results] The frequencies of rs10181656 C/G were 36.45 ％,46.54％ in genotype C/C,46.99％,45.38％ in genotype C/G and 16.57％,8.08％ in genotype G/G between URSA patients and normal controls.They reached statistical difference (P ＜ 0.05).The carriers of rs 10181656 G allele increased the risk of URSA (OR =1.50,P ＜ 0.05).STAT4 protein expression in decidual tissues:①There was statistical difference in the STAT4 protein expression in decidual tissues between cases and controls (P ＜ 0.05).In URSA patients there was statistical difference in the STAT4 protein expression among genotype CC,CG and GG of rs10181656 locus (P ＜ 0.05).So was in normal controls (P ＜ 0.05).In genotype CC there was no difference in the STAT4 protein expression between cases and controls (P ＞ 0.05).Neither was In genotype CG and GG respectively (P all ＞ 0.05).[Conclusion] Functional polymorphisms of the rs10181656 locus could probably associate with the susceptibility of URSA via STAT4 protein expression increased by genotype G/G in maternal decidual tissue.

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α(+) dendritic cells (DCs) in vitro was investigated in this study. Immature CD8α(+) DCs were prepared from C57BL/6 (H-2(b)) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α(+) DCs were pulsed by allogeneic (Balb/c, H-2(d)) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α(+) DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α(+) DCs and T co-culture supernatant were detected by using ELISA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced from C57BL/6 (H-2(b)) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α(+) DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α(+) DC/T ratio increased (P<0.05). When antigen concentration ≤ 5 μg/mL and CD8α(+) DC/T ratio ≤ 2:1, the ability of CD8α(+) DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P<0.05), but not in syngeneic tumor antigen-pulsed groups (P>0.05). The level of IFN-γ and IL-10 in CD8α(+) DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P<0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α(+) DC/T was 1:1 or 2:1 (P<0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxicity assay showed that when CD8α(+) DCs were pulsed with allogeneic tumor antigen, the maximal T cell killing efficiency could reach (100±7.7)%, whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)%. It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α(+) DCs could stimulate T cells to exert the GVT effect in vitro, and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen. The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α(+) DC/T ratio (1:1 and 2:1).

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α+dendritic cells (DCs) in vitro was investigated in this study.Immature CD8α+ DCs were prepared from C57BL/6 (H-2b) bone marrow cells by using a cytokine cocktail.On the 3rd day of culture,CD8α- DCs were pulsed by allogeneic (Balb/c,H-2d) EL9611 leukemia antigen,or RM-1 syngeneic prostate cancer antigen,with the concentration series of 0,2.5,5.0,10.0,20.0 μg/mL,respectively,then antigen-loaded immature CD8α+ DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1∶1,2∶1and 4∶1.T cell proliferation was measured by MTT assay.Cytokines including interferon gamma (IFN-γ)and interleukin-10 (IL-10) in CD8α+ DCs and T co-culture supernatant were detected by using ELISA.Cytotoxic effect of antigen-specific T cells was tested by LDH release assay.Conventional mature DCs　(mDCs) induced from C57BL/6 (H-2b) bone marrow cells by using granulocyte-macrophage colony　stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control.The results showed that the　proliferative activity of T cells stimulated by CD8α+ DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α+ DC/T ratio increased (P＜0.05).When antigen concentration ≤ 5μg/mL and CD8α+ DC/T ratio ≤ 2∶1,the ability of CD8α+ DCs to stimulate T cell proliferation was　higher than mDC control in allogeneic tumor antigen-pulsed groups (P＜0.05),but not in syngeneic tumor antigen-pulsed groups (P＞0.05).The level of IFN-γ and IL-10 in CD8α+DCs and T cell co-culture　supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P＜0.05),and the　cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when　the CD8α+DC/T was 1∶1 or 2∶1 (P＜0.05).There existed a negative correlation between the level of　IL-10 and T cell proliferation.T cell cytotoxicity assay showed that when CD8α+ DCs were pulsed with　allogeneic tumor antigen,the maximal T cell killing efficiency could reach (100±7.7)％,whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)％.It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α+ DCs could stimulate T cells to exert the GVT effect in vitro,and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen.The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α+ DC/T　ratio (1∶1 and 2∶1).