Objective To investigate the effect and mechanism of metformin on endometrial carcinoma subline of progestin-resistance and find whether metformin could regulate progestin-resistance in endometrial carcinoma. Methods Ishikawa endometrial carcinoma cells were cultured for a long period in the presence of the synthetic medroxyprogesterone 17-acetate (MPA) to generate a subline refractory to the growth-suppressive effects of MPA,named Ishikawa/MPA cells. The effect of MPA (1, 5, 10, 20, 40 and 60 μmol/L) or metformin (1, 10, 20, 40, 60, 70 and 80 mmol/L) on proliferation of the Ishikawa/MPA and parental Ishikawa cells was detected by cell counting kit-8 (CCK-8) method. Western blot was used to detect the effect of metformin and(or)MPA on the expression of PR-B in the Ishikawa/MPA and Ishikawa cells. Results The Ishikawa/MPA showed that growth stimulation rather than inhibition in the Ishikawa cells after low MPA concentration treatment. The doubling time of Ishikawa/MPA cell lines were (43±4) hours and that of Ishikawa cell line were (47 ± 3) hours. No changes in doubling time were observed (t=0.349,P=0.572). Low concentration (1 and 5μmol/L) of MPA could promote the growth of Ishikawa/MPA cells (by 3% and 13%). High concentration (10, 20, 40 and 60 μmol/L) of MPA could inhibited the growth of Ishikawa/MPA cells (by 4%, 3%, 9%and 40%) and the growth of Ishikawa cells (by 41%, 55%, 65%and 66%). At the same concentration, the difference of inhibition rates between the two cell lines were statistically significant (P<0.01). Metformin (1, 10, 20, 40, 60, 70 and 80 mmol/L) could inhibite the growth of Ishikawa/MPA (by-10%, 20%, 56%, 89%, 97%, 98%and 99%) greater than those in parental Ishikawa cells (by-6%, 19%, 37%, 54%, 70%, 72%and 83%), and the inhibitory effect was dose-dependent manner. Western blot assay showed that for Ishikawa cells, the protein expression levels of PR-B in metformin group and MPA+metformin group were respectively (53.5±4.0)%and (37.7±5.2)%, which were higher than that in the control group [(23.4 ± 3.0)%], and there were significant the differences (P<0.01). For Ishikawa/MPA cells, the protein expression levels of PR-B in metformin group and MPA+metformin group were respectively (38.6 ± 1.7)%,(36.3 ± 2.5)%,which were higher than those in the control group [(6.4 ± 1.6)%], and there were also significant differences (P<0.01). Conclusion Metformin may regulate the progestin-resistance in endometrial carcinoma by increasing the expression of PR-B.

Objective To investigate the urodynamic measurements in later pregnant women with urinary incontinence. Methods According to the symptoms, a total of 63 volunteers in later pregnancy were divided into two groups including urinary incontinence group and no symptom group. Fourteen women who were married but not delivered were included in control group. Urodynamic study was performed on all women. Results The occurrence rate of urinary incontinence in later pregnancy was 26.98%. The valsalva leak point pressure only occurred on two pregnant women were 50 cmH2O and 67 cmH2O respectively. Compared with the no symptom group, the maximum urethral closure pressure[(83.69±42.55)mmHg vs(108.09±34.95)mmHg, P＜0.05])and the functional urethral length [(30.45±8.42)mm vs (37.60±18.45)mm ,P＜0.05]of urinary incontinence group were decreased obviously. Conclusions The main reason of urinary incontinence in pregnancy was that the maximum urethral closure pressure could not sufficiently increase to compensate for the progressive increase in bladder pressure during pregnancy and functional urethral length could not correspondingly increase along with the pregnancy.

Nursing experience in the therapy of 45 cases of Rh blood group incompatibility was reported,and the nursing requirements were raised as follows:(1)good nurse patients relationship is important and psychological nursing should be done well.(2)Teach the patient to insist of systematic treatment,master timely intrauterine state of fetus.(3)Observe progressions and regression of jaundice,and general statement of newborn.(4)Enhance newborn nursing in the phototherapy.(5)Observe newborns physical signs in the course of blood exchange,predict the occurrence of complications.Results:one case was labor induced sine fetal death,40 cases lived without complications,4 cases died,the cure rate reached 88.9 percent.Conclusion:Though Rh blood group incompatibility is severe,most of them can be cured through timely treatment and careful nursing.

Objective To study telomerase activity in cervical cancer and it′s precursor lesion Methods Thirty six cervical cancer and 16 cervical intraepithelial neoplasia (CIN) specimens were measured for telomerase activity using TRAP ELISA, and 11 normal cervix, 6 chronic cervicitis and 8 adjacent normal tissue specimens as controls Results Mean telomerase activity in CIN, cervical cancer, and controls were 0 398?0 293, 1 580?0 819, 0 050?0 012 There was statistically significant difference among three groups ( P

Objective To explore the expression and function of myosin light streptokinase (MLCK) in small intestine mucosa of acute necrotizing pancreatitis (ANP) rats.Methods Fifty-six male SD rats were randomly assigned to control group and ANP group.A rat model of ANP was reproduced by retrograde injection of 4％ sodium taurocholate into the biliopancreatic duct,while the control group underwent a sham operation.The rats were sacrificed at 6th,12th,24th,48th hour after ANP induction.Serum amylase、TNF α,IL 1β,diamine oxidase (DAO) were measured.The pathological scores in the pancreas and small intestine were observed.The ultrastructure and tight junction (TJ) changes in the small intestine mucosa were observed with an electron microscope.The localization and expression of MLCK in small intestine mucosa was determined by immunohistochemistry method.Results Compared to the control group,the serum amylase,TNF-α,IL-1 β,DAO level,in the ANP group were all significantly increased;[(4 978 ± 1 574) U/L vs (1 176 ± 124))U/L,(47.88 ± 15.85) μg/L vs (17.24 ± 1.99) μg/L,(132.48 ± 68.54) μg/L vs (23.51 ± 6.44) μg/L,(95.96 ± 30.84)μg/L vs (38.06 ± 17.73)U/L at 12 h],and the pathology scores of pancreas and small intestine were both significantly elevated [12 h:(12.2 ± 1.80) vs (4.68 ± 0.35),(2.58 ± 0.52) vs (0.58 ±0.26)] (P ＜0.05);the MLCK protein expression in small intestine mucosa was significantly increased in ANP group (12 h:0.1863 ± 0.0230 vs 0.1636 ± 0.0049),and the difference was statistically significant (P ＜0.05).The small intestine ultrastructure was seriously damaged and TJ was widened significantly in ANP Group.Conclusions The increased serum TNF alpha and IL-1β concentration and DAO activity and up-regulated MLCK protein expression in small intestine mucosa may damage the integrity of tight junction of intestinal epithelial cell and cause intestine mucosa barrier dysfunction.

Objective To analyze the mutations of IDS gene in a mucopolysaccharidosis type Ⅱ (MPS Ⅱ) family and to make prenatal diagnosis on the high-risk fetus which has been pregnant for eleven weeks.Methods IDS gene was analyzed by bidirectional DNA sequencing in 2 patients and their mother,and 5 unaffected individuals.Prenatal diagnosis for the high-risk fetus was performed by chorionic villus sampling after the genotypes was identified.Results The mutation c.344delA (N115fsX15) was detected in the two patients,and the mother of patients carried the heterozygous c.344delA (N115fsX15) mutation.None of the mutant was detected in the 5 unaffected subjects.The fetus carried c.344delA (N115fsX15) heterozygous mutation and was a carrier.Conclusion The deletion mutation c.344delA (N115fsX15) is causative to the pedigree of MPS Ⅱ,and prenatal diagnosis is the efficient method to avoid defect birth.

Objective To investigate the effects of metformin on cell proliferation in differentiation degree of endometrial carcinoma cells and related mechanisms. Methods The endometrial cancer cell lines Ishikawa and AN3CA were used. Cell proliferation was assessed after exposure to metformin with or without epithelial growth factor receptor (EGFR) inhibitor AG1478 by cell counting kit-8 (CCK-8) method. EGFR mRNA was determined by reverse transcription (RT)-PCR. The expression of phosphorylation EGFR (p-EGFR) and total EGFR (t-EGFR) and phosphorylation extracellular signal-regulated kinase 1/2 (p-ERK1/2) and total ERK1/2 (t-ERK1/2) were examined by western blot. Results (1)CCK-8 experiment showed that metformin could inhibit the proliferation of endometrial cancer cells in a time-dependent manner and a dose-dependent manner (P<0.05), but the inhibition of well differentiated cell line Ishikawa was lower than that in poorly differentiated cells AN3CA (P<0.05). AG1478 also could inhibit the proliferation of endometrial cancer cells in a time-dependent manner and in a dose-dependent manner (P<0.05), but the inhibition rate of well differentiated cell line Ishikawa was higher than that in poorly differentiated cells AN3CA (P<0.05). Metformin+AG1478 also could inhibit the proliferation of endometrial cancer cells in a time-dependent manner and in a dose-dependent manner (P<0.05), and the inhibition of combined with metformin and AG1478 was stronger than that with a single application of drugs, but the inhibition rate of Ishikawa was higher than that in AN3CA (P<0.05).(2)RT-PCR method showed that different concentrations of metformin (0.01, 0.1, 1, 5, 10 mmol/L, respectively) for 24 hours, the expression level of EGFR mRNA in Ishikawa cells were respectively 0.74±0.03, 0.61±0.04, 0.46±0.03, 0.31±0.03 and 0.23±0.03, the expression level of EGFR mRNA in AN3CA cells were respectively 0.79±0.20, 0.61±0.03, 0.50±0.05, 0.32±0.03 and 0.26 ± 0.04, the inhibition effect showed a significant concentration-dependent manner (all P<0.01). (3) Western blot method displayed that the effect of metformin treated respectively 2, 4, 6 or 8 hours, there were not significant difference in the expression levels of t-EGFR protein and t-ERK1/2 between Ishikawa and AN3CA cells (all P>0.05). But the expression levels of p-EGFR and p-ERK1/2 protein were significantly lower between two groups (P<0.01), which showed a time-dependent manner(P<0.01). Conclusion Metformin could inhibit the proliferation of endometrial cancer cells, the inhibition is associated with the differentiation degree of cancer cells. Metformin could enhance the EGFR signaling pathway inhibitor AG1478 inhibition of endometrial cancer cells, which may inhibit EGFR expression of phosphorylated proteins to inhibit the phosphorylation of ERK1/2 proteins and then inhibit proliferation of endometrial cancer cells.

Objective To investigate the effects of signal pathway inhibitors PD98059 and LY294002 on cell proliferation, apoptosis, expressions of phosphorylated extracellular signal-regulared kinase (p-ERK) and phosphorylated protein kinase B ( p-Akt) in endometrial carcinoma xenografts. Methods Human endometrial carcinoma Ishikawa cells were cultured in vitro. The effects of PD98059 and LY294002 on proliferation, apoptosis, and cell cycle distribution of endometrial cancer cells were detected by monotetrazolium ( MTT) assay and fluorescence-activated cell sorting technique. The models of xenografted tumor were established by the subcutaneous inoculation in 24 nude mice, and then they were randomly divided into 4 groups ( n = 6) , normal saline group, PD98059 group (PD group) , LY294002 group ( LY group) or PD98059 + LY294002 group ( PD + LY group) by intraperitoneal injections, respectively. The anti-tumor efficacy was evaluated by measuring tumor volume and tumor growth status. The histopathological change of tumor specimens was observed using HE staining and terminal deoxynucleotidyl transferasemediated dUTP-digoxigen in nick and labeling method (TUNEL) testing and the expression levels of p-ERK and p-Akt were detected by immunohistochemistry method. Results ( 1) The proliferation of Ishikawa cells were suppressed after treated by PD98059 and ( or) Y294002, in which A570 values of cells decreased showing both time-dependent and concentration-dependent manner ( LY294002: Fgroup = 9. 801, P = 0. 002; Ftime = 10. 398, P = 0. 001. PD98059: Fgroup= 8. 213, P = 0. 015; Ftime = 6. 839, P = 0. 036). Cell cycle distribution analysis revealed that percentage of Ishikawa cells at G0/G1 phase(Ftime =35.049, P= 0.004; Fgroup = 32. 024, P ＜0. 01) increased and percentage of S phase cells (Ftime = 7. 789, P = 0. 049; Fgroup = 30. 132, P ＜0. 01) decreased significantly. The percentage of apoptotic cells increased significantly among PD group, LY group and PD + LY group, in which there were significant difference [(63. 3 ±0.5)% vs (30. 7 ± 20. 1) % vs(40. 8 ± 1. 3) % ; F = 621. 059, P ＜ 0. 01]. (2) Compared with the control group, the increasing of transplanting tumor volume in the treated groups were obviously ( F = 23. 545 , P ＜ 0. 01) , and the inhibited rate of the tumor was higher in PD + LY group than that in PD group or LY group [(68 ± 9 ) % vs ( 32 ± 16 ) % or ( 38 ± 17 ) % ; F = 10. 283 , P ＜ 0. 05]. ( 3 ) HE staining shown that there were different degrees of necrosis for endometrial carcinoma cell in different groups. The apoptosis of tumor cells were significantly increased in treated groups by TUNEL testing [(13. 7 ± 1. 5)% , ( 14. 1 ± 1. 2)% , (29. 0 ± 1. 8 ) % ; F = 320. 344, P ＜ 0. 01]. Immunohistochemistry results demonstrated that the expressions of p-ERK and p-Akt in treated groups were lower than that in control group, of which LY + PD group was the lowest one. Conclusion The signal pathway inhibitors PD98059 and LY294002 could inhibit the growth of human endometrial carcinoma in vivo and in vitro, in which may induce cell apoptosis.

BACKGROUND: Treatment of ulcerative colitis (UC) with herbs-partition moxibustion is of good intervenient effect, whether the effect is related to neutrophil apoptosis in the pathogenic process of UC?OBJECTIVE: To investigate the changes of neutrophil appptosis rate of rats with UC and the regulative effect of medicinal cake moxibustion. DESIGN: A randomized controlled trial based on SD rats.SETTING: Shanghai Research Institute of Acupuncture and Meridian.MATERIALS: The experiment was completed from August 2002 to June 2003. A total of 30 healthy male SD rats of clean grade, weighing (140±20) g,provided by Experimental Animal Center of Shanghai University of Traditional Chinese Medicine, were at random divided into 2 groups: model building group (n=20) and normal control group (n=10).METHODS: Rat UC models were set up with immunological method. After the models were successively set up, four model rats and two normal rats were randomly selected for pathological observation of distal colon tissue. The rest models were divided as model group and herbs-partition moxibustion group with 8 rats for each, and the rest 8 normal rats were taken as control group. The rats of herbs-partition moxibustion group were given herbs-partition moxibustion at tian shu (ST-25) of both sides and qi hai (CV-6) points; the herbs cones was made from Radix Aeoniti Praeparata,Cortex Cinnamoni, and so on. About 90 mg moxa cone was put on the herbs cones for moxibustion, once a day, for 10 times. After treatment, all rats of the three group were executed, then the peripheral monocytes were isolated for cell culture. A 1:50 diluted supernatants of each group were taken, and the same volume of RPMI-1640 culture fluid was used as blank control group. They were respectively incubated together with neutrophils of peripheral blood of normal rats. The neutrophil apoptosis rates were measured by using flow cytometry.MAIN OUTCOME MEASURES: Comparison of neutrophil apoptosis rates of rats in each group.RESULTS: Totally 20 rats in model group and 10 in normal control group were accepted. Then 4 rats in model group and 2 in normal group were sacrificed and observed with tissue pathology, and 16 and 8 rats entered the final analysis in model and normal control groups respectivelly. ① The neutrophil apoptosis rate of UC rats was significantly lower that that of rats in blank control group [(16.34±2.80) ％, (52.33±9.94) ％, q= -35.99, P＜ 0.01]; but there was no significant diffenence between normal control group [(48.79±11.33) ％] and blank control group, (P＞0.05). ② After treatment the neutrophil apoptosis rate of herbs-partition moxibustion group [(36.03±8.31) ％] was significantly higher than that of UC model group (q=19.69, P ＜ 0.01), but it was still lower than that of normal control group (q= -16.30, P ＜ 0.05).CONCLUSION: One of the main mechanism for herbs-partition moxibustion to treat UC might be regulating and/or reducing the inhibited state of neutrophil apoptosis in UC.

Objective To investigate therapeutic effect of the low dose clomiphene treatment of idiopathic oligoasthenospermia and effect on sperm epididymal protein level 4.Methods183 cases of idiopathic oligoasthenospermia were divided into three groups, vitamin group of 61 patients were given vitamin E,clomiphene group(61 cases)treated with low dose clomiphene, combined group of 61 patients were given low dose clomiphene and vitamin E, lasted for 12 weeks.Semen volume, sperm density, sperm motility, sperm survival rate and sperm reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), human epididymis protein 4 (HE4), follicle stimulating hormone, luteinizing hormone, testosterone concentration were observed before and after treatment.After treatment, the patients were followed up for 1 year, the observation of the natural pregnancy rate after treatment.ResultsSperm density, sperm survival rate and sperm and a A+B grade sperm in the clomiphene group were significantly higher comparedwith the vitamin group (P< 0.05), sperm density, sperm survival rate and sperm and a A+B grade sperm in the combined group were significantly increasedcompared with clomiphene group and vitamin group (P< 0.05),levels of HE4 after treatment was significantly lower compared with before treatment (P< 0.05), And the difference between the three groups were statistically significant (P< 0.05).Follicle stimulating hormone, luteinizing hormone, testosterone in clomiphene group and combined group after treatment, were significantly increased compared with before treatment (P< 0.05), and the combined group had no statistically difference compared with clomiphene group.The pregnancy rate in 1 year of vitamin group, clomiphene group and combined group were 24.59%, 42.62% and 62.29% (P< 0.05).ConclusionClomiphene treatment of idiopathic asthenospermia is better than vitamin E, the combination of the two has a synergistic effect.