OBJECTIVE: To evaluate the efficacy of Marvelon combined with metformin in the treatment of polycystic ovary syndrome(PCOS). METHODS: Both prospective and retrospective clinical studies on the efficacy of Marvelon combined with metformin were searched in several databases of medicine and pharmacy, and the data was analyzed by evidence-based literature analysis method. RESULTS: There were nine randomized controlled trials(RCTs) in Chinese on Marvelon combined with metformin for the treatment of PCOS. Marvelon combined with metformin could improve hormone levels, promote ovulation and pregnancy rate markedly. CONCLUSION: It has been demonstrated in the researches that Marvelon combined with metformin is more effective for polycystic ovary syndrome(PCOS), however, further randomized, double-blind studies with larger sample size are needed to prove the efficacy and safety of Marvelon combined with metformin.

Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.

Objective To share our experiences on integrated services in providing fetal diagnosis and postnatal treatment for congenital diaphragmatic hernia(CDH).Methods A retrospective analysis was conducted on 25 pregnancies diagnosed as CDH by both prenatal ultrasound and MRI in Maternal and Children Hospital of Guangdong Province from January 2012 to January 2014.All of the subjects received integral medical management including prenatal management (prenatal diagnosis and consultation), perinatal management (prenatal care and delivery) and neonatal treatment.Results Among the 25 CDH fetuses, 11 were mild, nine were moderate, and five were severe.One severe case, who was diagnosed at 26 gestational weeks, was aborted on demand of the mother.The other 24 cases continued their pregnancy and all delivered after 35 weeks including 13 cesarean sections (one due to twin pregnancy and 12 due to maternal demand) and 11 vaginal birth.The mean gestational age when CDH was diagnosed was (24.5 ± 3.5) weeks, and the 24 women delivered at an average of (37.5 ± 1.4) gestational weeks.The eleven mild cases accepted mask oxygenation.For those 13 moderate or severe CDH cases, all received dexamethasone to promote fetal lung maturity at 32 gestational weeks, seven were intubated before clamp the cord, and the other six did after.These 13 babies accepted high-frequency oscillation ventilation, with a median duration of 58 hours, and some of them treated with inhaled nitric oxide on requirement with a median duration of 52 hours.Except two cases died before operation, the rest 22 cases underwent neonatal surgery.One moderate case died at 48 hours after surgery due to pulmonary hypertension and respiratory failure.Another one severe case withdrew treatment at two months old.The other 20 infants recovered fully.Conclusions Integrated management including prenatal diagnosis and postnatal treatment, provides an effective and streamlined mode for diagnosis and treatment of CDH.Therefore,it might minimize potential medical risks.

AIM: Using optical coherence tomography angiography (OCTA) to observe the changes and clinical significance of macular blood flow density in patients with diabetic retinopathy (DR).METHODS: Totally 47 eyes (28 patients) with diabetic retinopathy (DR) were enrolled in the DR group.According to the international clinical grading criteria of diabetic retinopathy, 30 eyes (19 patients) with non-proliferative diabetic retinopathy were classified as the NPDR group, and 17 eyes (11 patients) with proliferative diabetic retinopathy were classified as PDR group.A total of 46 (27 subjects) healthy eyes with matched age were enrolled in the control group.All the subjects underwent the 3mm×3mm scanning of macular retina by optical coherence tomography angiography (OCTA), obtaining 4 levels of macular blood flow density map.The macular blood flow density at 3 levels, including superficial retinal layer, deep retinal layer and choroidal capillaries layer, were measured.RESULTS: The macular blood flow density of superfical retinal layer, deep retinal layer and choroidal capillaries layer in DR group were 0.4963±0.0840, 0.4798±0.0801 and 0.5290±0.0528, respectively.Among them, the blood flow density of each layer were 0.5064±0.0843,0.4983±0.0766,0.5345±0.0529, respectively, for the NPDR group, and were 0.4786±0.0830, 0.4473±0.0778,0.5192±0.0526, respectively, for the PDR group.For the control group, the density of each layers were 0.5919±0.0704, 0.6301±0.0527, 0.5691±0.0169, respectively.The macular blood flow density was significantly different in the superficial retinal layer, deep retinal layer and choroidal capillary layer between the control group and the NPDR group, as well as the PDR group and the DR group (total P<0.001).Statistically significant difference was found between the NPDR group and the PDR group in the deep retina layer (P=0.029), but not in the superficial retina layer and choroid capillary layer (P=0.236, 0.268).CONCLUSION: Compared with the control group, the macular blood flow density of superficial retinal layer, deep retinal layer and choroidal capillary layer in the patients with diabetic retinopathy decreased significantly.It indicated that the macular ischemia existed in both retina and choroid.By quantitatively measurement of the macular blood flow, OCTA may be used for monitoring the progression of diabetes, and early detection of diabetic retinopathy.

Background In vitro study showed that chemotaxis consist of chemotaxis factor 4(CXCR4)and stromal cells derived factor-1(SDF-1)and may play a role in the orientation and migration of retinal progenitor cells (RPCs)toward lesion.Overexpression of CXCR4 in RPCs can enhance the chemotaxis activity.Objective This work was to explore the feasibility and underlying mechanism of up-regulation of CXCR4 on RPCs induced by hypoxia.Methods RPCs were retained in an incubator with normal O2volume(16％)or hypoxia condition(10％ O2)for 12 hours and 24 hours respectively.Flow cytometer cell analysis screening(FACS)was conduced to measure the proportion of CXCR4-expressing cells,and CXCR4,HIF-1 mRNA were analyzed by reverse transcription-polymerse chain reaction(RT-PCR).The chemotical effect of 30 mg/L SDF-1 to RPCs cultured under the hypoxia condition was assessed using Boyden chamber.Results The expression level of CXCR4(CXCR4 mRNA/β-actin mRNA)inRPCs cultured by 10％ O2 for 12 and 24 hours were 0.28+0.07and 0.48+0.17 and increased by 1.75 and 3.00 fold more than that of 16％ O2 culture group(0.16+0.02)(P＜0.01).The expression level of HIF-1 mRNA(HIF-1 mRNA/β-actin mRNA)in RPCs cultured by 10％ O2 for 12 and 24 hours were 0.18 ±0.07and 0.38 ±0.13 and increased by 3.00 and 6.30 fold more than that of 16％ O2 culture group(0.06±0.01)(P＜0.01).The chemotical effect of 30 μg/L SDF-1 to RPCs increased from 13.00％ in 16％ O2 culture group to 36.00％ and 46.00％ in the cells cultured by 10％ O2for 12 and 24 hours.FACS revealed that the proportion of CXCR4+ cells in hypoxia-exposure for 12 and 24 hours were 26.90％ and 46.10％,respectively,but that in 16％ O2 culture group was 9.10％,showing a statistically significant difference(P ＜ 0.01).Conclusions RPCs induced by hypoxia can enhance the expression of CXCR4 in RPE cells and the chemotaxia to SDF-1.The overexpression of H1F-1 in RPCs may be involved in the up-regulation of CXCR4 expression.

Aim To investigate the up-regulation of miR-22 through Wnt pathway inhibits the proliferation,migration and invasion in human gastric MGC803 cells induced by diallyl disulfide(DADS).Methods The effects of proliferation,migration,and invasion of gastric cancer cells were evaluated by MTT,wound-healing and invasion assays.Online prediction software was applied to search the target gene of miR-22.Luciferase report gene assay was used to assess the target genes Wnt-1 of miR-22.The expressions of Wnt-1,β-catenin and TCF-4 were tested by qRT-PCR and Western blot,respectively.Results MTT showed that DADS and miR-22 notably decreased the proliferation compared with control group(P<0.05).Wound-healing assay showed that DADS and miR-22 could significantly inhibit the migration of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Invasion assay showed that DADS and miR-22 could markedly inhibit the invasion of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Online prediction software to search the target gene exhibited that Wnt-1 may be a target gene of miR-22. Luciferase report gene assay disclosed that Wnt-1 was identified as a direct target of miR-22. Qrt-PCR showed that the expression of Wnt-1 Mrna was respectively down-regulated by DADS and miR-22 compared withcontrol group, especially in miR-22+DADS(P<0.05). Western blot exhibited that DADS and miR-22 obviously suppressed the expressions of Wnt-1, β-catenin and TCF-4 proteins, especially in miR-22+DADS(P<0.05).Conclusion Up-regulation of miR-22 through Wnt pathway can remarkably suppress the proliferation, migration and invasion in MGC803 cells by DADS.

Animals ; Cell Line, Tumor ; Enoyl-CoA Hydratase ; genetics ; metabolism ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Lymphatic Metastasis ; Mice ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To discuss the application of prospective risk nursing measures in reducing the occurrence of postoperative local hematoma at femoral artery puncture site. Methods A total of 605 patients receiving femoral artery puncture for interventional management were collected as the control group , and 322 patients were used as the intervention group. The risk factors causing postoperative local hematoma at femoral artery puncture site in the control group were recorded, and based on which risk estimation survey table was designed. According to risk estimation survey, the prospective intervention measures, including risk assessment, standardization of professional training, individual health education, the improvement of care appliance, etc. were carried out for the patients of the intervention group. The occurrences of postoperative local hematoma at femoral artery puncture site were recorded and the results were compared between the two groups. Results No statistically significant differences in demographic and clinical data existed between the two groups. However, the incidence of hematoma in the intervention group was significantly lower than that in the control group (χ2 = 4.652, P < 0.05), although the difference in the severity of hematoma was not significant between the two groups. Conclusion The use of prospective risk nursing measures can effectively reduce the incidence of postoperative hematoma at femoral artery puncture site.

Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.

Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.