ve To determine the most approprite hypoxic concentration and duration for prenatal hypoxic adaptation animal experiment by exposing pregnant rats to the hypoxic air of different oxygen concentration.Methods Full-term pregnant rats( gestation time 22 days) were placed in an airtight cabin specially designed for hypoxic adaptation experiment. The rats were divided into 7 groups. The Q2 concentration in the airtight cabin was decreased from 21% (group Ⅰ as control) to 18% (group Ⅱ), 17% (group Ⅲ), 16% (group Ⅳ), 15% (group Ⅴ), 14% (group Ⅵ) and 13% (group Ⅶ) respectively. The animals were exposed to short duration of hypoxic air twice with a break of 5min breathing fresh air. The duration of the first hypoxic episode lasted 10 min (group Ⅰ ) , 5 min (group Ⅱ), 7.5min (group Ⅲ), 9.83 min (group Ⅳ), 11.5 min (group Ⅴ), 13.17 min (group Ⅵ) and 14 min (group Ⅶ) respectively. The second hypoxic episode lasted 10min, 9.33 min, 11 min, 15.17 min, 13.33 min, 17 min and 18 min respectively. Ten newborn rats (1 day after birth) randomly selected from each group were placed in a 100ml airtight bottle and the duration from the start to the time when the newborn rat stopped breathing was recorded as hypoxia surviving time. Another 10 newborn rats randomly selected from each group were decapitated and brain was removed for light and electron microscopic examination to determine the degree of neuronal damages. Results In group Ⅰ-Ⅴ the newborn rats were normal (pink skin color and good extremity movement) . In group VI 10/55 (18%) newborn rats were cyanotic with diminished extremity movement, the others were normal. In group VIII 11/52(21% ) newborn rats died, 14/ 52(27%) were cyanotic with diminished extremity movement. Neuronal damages could be seen in cyanoticnewborn rats including decreased number, swelling, apoptosis of neurons and expanded mitochondria. The hypoxia surviving time was significantly longer in group Ⅳ, Ⅴ and Ⅵ than that in control group. Conclusions Hypoxic air containing 15% O2 is appropriate for animal experiment of prenatal hypoxic adaptation. It is better to divide prenatal hypoxia into two episodes lasting 11.5 min and 13.33 min with a break of 5 min between them when animals breathe fresh air.

Angioplasty, Balloon, Coronary ; Humans ; Percutaneous Coronary Intervention ; methods ; Tungsten

Objective To study the role and mechanism of microRNA-16(miR-16)in the proliferation,invasion and apoptosis of ovarian epithelial carcinoma cells in vitro.Methods The SKOV-3 cells were transfected with miR-16 mimics or negative control RNA(NC)by lipofectamine 2000.The expression of miR-16 was detected by real-time reverse transcription(RT)-PCR in SKOV-3 cells,and western blot was used to detect the expression of vascular endothelial growth factor(VEGF),matrix metalloproteinase-2(MMP-2)and bcl-2 protein.Methyl thiazolyl tetrazolium(MTT),5-ethynyl-2'-deoxyuridine(EdU)and transwell assay were used to determine the proliferation and invasion abilities.And the rate of apoptotic cell was detected by flow cytometry method.Results(l)The expression level of miR16 in the transfection cells group was significantly higher than that in NC group(125.93 ± 15.30 versus 0.78 ± 0.16,P ＜ 0.01).(2)The rclative expression level of VEGF protein in transfection cells,NC and blank control group was 0.58 ± 0.05,1.22 ± 0.03,1.20 ± 0.03,MMP-2 protein was 0.63 ± 0.03,1.16 ±0.03,1.21 ± 0.03,and bel-2 protein 0.52 ± 0.03,1.19 ± 0.05,1.28 ± 0.06,respectively.The level of VEGF,MMP-2 and bcl-2 protein in the transfection group were lower than those in other control groups,and there were significantly differences among them(all P ＜0.01).(3)After transfected 4 days,the inhibition rate of cell proliferation in the transfection group was dramatically higher than that in NC group[(37.2 ±6.2)％ versus(3.6 ± 3.2)％,P =0.001].(4)The percentage rate of proliferative cells in the transfection,NC and blank control group was(12.3 ± 0.8)％,(23.4 ± 1.8)％,(31.1 ± 4.9)％.And it was lower in the transfection group(P ＜ 0.05).(5)Decreased cells via the transwell member in the transfection group(6 ± 3)were detected as compared with NC group(40 ± 9)and blank control group (48 ± 8,P ＜ 0.01).(6)Twenty-four hours after cultured in serum starvation and hypoxia,the rate of the viable and late apoptotic cells in the transfection group were significantly higher than those in NC group and blank control group[the rate of viable apoptotic cell was(16.9 ± 2.1)％,(10.3 ± 1.7)％ and(9.0 ±0.8)％ respectivcly,P＜0.01;the rate of late apoptotic cell was(13.4±3.3)％,(3.2 ±1.8)％ and (0.7 ±0.6)％ respectively,P ＜ 0.01].After cultured 48 hours,total apoptotic cells in the transfection group was significantly more than those in other groups(P ＜ 0.01).Conclusion miR-16 might inhibit the proliferation,invasion of ovarian epithelial carcinoma cells and enhance their sensitivity to apoptotic stimuli via downregulation of the expression of VEGF,MMP-2 and bcl-2 protein.

Objective To investigate the expression of long intergenic non-protein coding RNA-regulator of reprogramming (Linc-ROR) in high-grade ovarian serous cancer, and explore the relationship between Linc-ROR expression and biological function of high-grade ovarian serous cancer. Methods A total of 34 high-grade ovarian serous cancer tissue samples and 19 normal fallopian tube tissue samples were collected between June 2014 and February 2016. Real-time reverse transcription (RT)-PCR was used to detect the Linc-ROR mRNA expression in different samples. The relationship between Linc-ROR expression level and ovarian cancer International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis was analyzed. Constructed Linc-ROR small interference RNA (siRNA) and pIRES2-EGFP-Linc-ROR plasmid, then Linc-ROR siRNA and pIRES2-EGFP-Linc-ROR plasmid were respectively transfected into SKOV3 cells. Cell proliferation, migration and invasion ability were assessed by cell counting kit-8 (CCK-8), wound healing assay and transwell invasion assay. Results (1) The expression level of Linc-ROR mRNA was significantly higher in high-grade ovarian serous cancer than normal fallopian tube tissues (4.31± 0.38 vs 1.03 ± 0.21; t=25.842, P<0.01). With the progression of FIGO stages, the expression of Linc-ROR was increased (F=95.702, P<0.01), and it was associated with lymph node metastasis (t=7.397, P<0.01). (2) The results of RT-PCR showed that the expression level of linc-ROR in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (0.30 ± 0.11 vs 1.02 ± 0.10; t=15.269, P<0.01). The expression level in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (8.90 ± 0.45 vs 1.03 ± 0.17;t=21.934, P<0.01). The CCK-8 assay showed that when the cells were cultured for 3, 4, 5 and 6 days, the A value in Linc-ROR-i group was significantly lower than that in Linc-ROR-NC-i group (P<0.05). And the A value in Linc-ROR-p group was significantly higher than that in Linc-ROR-NC-p group (P<0.05). Wound healing assay showed that, after 48 hours incubation, migration rate of cells in Linc-ROR-i group was significantly less than that in the Linc-ROR-NC-i group [(52±4)%vs(67±5)%;t=5.720,P<0.01]. The migration of cells in Linc-ROR-p group was significantly greater than that in the Linc-ROR-NC-p group [(84±4)%vs(66±4)%;t=7.330,P<0.01]. Cell transwell invasion assay showed that, after 48 hours of incubation, the number of invasive cells in Linc-ROR-i group was lower than that in Linc-ROR-NC-i group (74 ± 3 vs 104 ± 3; t=15.810,P<0.01). And the number of invasive cells in Linc-ROR-p group was higher than that in Linc-ROR-NC-p group (217 ± 4 vs 108 ± 5; t=38.060, P<0.01). Conclusion Highly expressed Linc-ROR could enhance the proliferation, migration and invasion ability of high-grade ovarian serous cancer cells, which may be one of the important molecules in the occurrence and development, invasion and metastasis of high-grade ovarian serous cancer.

OBJECTIVE:To observe the efficacy and safety of paclitaxel combined with cisplatin and fluorouracil for gastric cancer with liver metastases via indwelling hepatic arterial catheter. METHODS:56 gastric cancer patients with liver metastases were randomly divided into control group(28 cases)and observation group(28 cases). Control group received Paclitaxel injection 135 mg/m2,d1+Cisplatin injection 75 mg/m2,d1+Fluorouracil injection 750 mg/m2,d1-5,pumping administrated via central venous. Ob-servation group received Paclitaxel injection 135 mg/m2,d1+Cisplatin injection 75 mg/m2,d1+Fluorouracil injection 750 mg/m2,d1-5, administrated via indwelling hepatic arterial catheter. 3-4 weeks were a course,it lasted 8 courses at most. Magnesium isoglycyrrhiz-inate injection 200 mg/d was intravenously infused for liver protection in 2 groups during treatment. Clinical efficacy,serum car-cinoembryonic antigen (CEA),alanine aminotransferase (ALT),aspartate aminotransferase (AST) levels before and after treat-ment,and the incidence of adverse reactions in 2 groups observed. RESULTS:Short-term clinical efficacy in observation group was significantly higher than control group,with statistical significance(P<0.01). Before treatment,there were no significant dif-ferences in CEA,ALT and AST levels(P>0.05). After treatment,CEA,ALT and AST levels in 2 groups were significantly high-er than before,ALT and AST levels in observation group were significantly higher than control group,while CEA level in observa-tion group was lower than control group,with statistical significances(P<0.05). The incidences of bone marrow suppression,nau-sea,vomiting and fever in observation group were significantly lower than control group,with statistical significances (P<0.05). CONCLUSIONS:Paclitaxel combined with cisplatin and fluorouracil has good efficacy for gastric cancer with liver metastases via indwelling hepatic arterial catheter,while it exists liver dysfunction.

objective To analyze the killer cell immunoglobulin-like receptors(KIR)gene polymorphism of pre-eclampsia patients and approach the correlation between KIR genes and pre-eclampsia.Methods The KIR gene polymorphisms of 71 pre-eclampsia patients and 100 healthy pregnant women were detected by PCR-SSP.The KIR2DL4 mRNA level in placentas from pre-eclampsia and gestational normal pregnancies were quantified by real time RT-PCR.Forty pre-eclampsia patients and 38 healthy pregnant women were detected for single nucleotide polymorphisms(SNP)in the gene coding and joint areas between introns and extrons by directly sequencing techniques of KIR2DL4 genomic DNA.Finally,all alleles and genotypes of KIR2DL4 gene were case-control studied.Result Distributions of some relatively activating KIR genotypes shewed a significant association with pre-eclampsia.Real-time RT-PCR showed that KIR2DL4 mRNA can be measured both in placenta of women with pre-eclampsia being of pre-eclampsia waft significantly lower than that of normal pregnancy.only as much as 0.276 times that of controls.We identified 18 polymorphisms,of which,7 were first reported.But no significant differences in genotype distributions or allele frequencies were observed in these SNPs between cases and controls.Conclusion The distributions of some relatively activating KIR genotypes showed a significant association with pre-eclampsia,which indicates that the polymorphism of KIR genes may be associated with the genetic predisposition to pre-eclampsia.And because the expression of KIR2DL4 mRNA in the placentas of cases wag significantly lower than control group,it is speculated that the decrease of KIR2DL4 expression in placenta may participate in the pathogenesis of pre-eclampsia.

Objective To investigate the clinic pathologic features of young patients with renal malignant tumour by comparing with older patients. Methods The clinical data of 83 younger adults(age,≤40 years) with renal cell carcinoma and 703 cases of older renal cell carcinoma (age＞40 years)was analyzed and compared retrospectively. All of the patients were surgically treated between 1986and 2007. Results Among patients with symptoms, younger female patients were more likely to have abdominal pain(12/27, 44.4% vs. 154/703, 21.9%)and mass (2/27, 7.4% vs. 154/703,1.3%)than older (P＜0.05). The older groups were more likely to have dear cell carcinoma than in the younger(501/703, 71.3% vs. 47/83, 56.6%)(P＜0.05). The rate of PRCC has the trends to be higher in young patients than that in older patients (21/83, 25.3% vs. 118/703, 16.8%) (P=0.054),especially in male patients (17/56, 30.4 %) (P=0.011). The rate of MFH and leiomyosarcoma was significantly higher in young patients than that in older patients (6/83, 7.2% vs. 12/703,1.7%) (P＜ 0.05), especially in female patients (4/27,14.8%). Younger male patients were more likely to have a higher incidence of small tumors (T≤4 cm)(24/56,42.9 % vs. 173/703,24.6 % )(P＜0.05)and a lower incidence of big RCC(T＞7 cm) than older patients(12/56,21.4% vs. 295/703,42.0%) (P＜0.05). Conclusion Younger female renal cell carcinoma patients tends to have worseprognosis than older patients, while younger male patients have better prognosis due to different pathology.

Objective To explore an effective evaluation method for students' self-and peer-assess-ment. Methods The students of 6 groups participating in extracurricular teaching activities were selected as research subject. Traditional method (final score = mean score of group/2 + teacher's score/2) and mean difference method [final score=teacher's score-(mean difference of group-mean difference of all groups)] were used to calculate final score of each group, and effect of two methods were compared. Results Scores of most groups were higher than the teacher's scores, and high scores were given by group 3 in self- and peer-assessment. The final score of all groups were higher than teacher's scores in traditional method. Compared with teacher's scores, final scores increased significantly in group 1, 4, 5 below mean difference, final score decreased significantly in group 2, 3 above mean difference, and final score did not differ in group 6 equal to mean difference in mean difference method. Conclusion The mean difference method can reflect the effect of student's self- and peer-assessment, and guide student to make objective and accurate evaluation. It is a more reasonable and scientific evaluation method for self-and peer-assessment.

ObjectiveTo evaluate the effect and safety of recombinant human erythropoietin (rhEPO) in treatment of lung cancer chemotherapy-related anemia.MethodsNinety-eight lung cancer chemotherapy-related anemia patients were divided into treatment group and control group with 49 cases each by random digits table method.The patients in treatment group were given rhEPO and chalybeate.The patients in control group were merely given chalybeate.The hemoglobin (Hb),hematocrit,allogeneic blood transfusion rate and quality of life between two groups were observed and compared.ResultsThree cases were rejected in treatment group,and 3 cases with anergy and dizzy and 2 cases with local injection site pain and sclerosis recovered spontaneously.Hb and hematocrit showed downward trend after treatment in control group,but there was no significant differences (P ＞ 0.05).Hb and hematocfit had upgrade trend after treatment in treatment group,and there were significant differences between after 4 - 8 months treatment and before treatment (P ＜ 0.05 ).The allogeneic blood transfusion rate was 24.5％ (12/49) in control group and 6.5％ (3/46) in treatment group,and there was significant difference between two groups (P ＜ 0.05 ).The quality of life in treatment group was increased compared with that in control group.There were significant differences in the effective rate after 4 or 8 weeks treatment between two groups [52.2％(24/46) vs.6.1％(3/49) and 95.7％ (44/46) vs.20.4％ ( 10/49 )].ConclusionsrhEPO is effective and safe in treatment of lung cancer chemotherapy-related anemia.rhEPO has little adverse reaction and can improve the quality of life.

ObjectiveTo investigate the role and mechanism of microRNA-21 (miR-21) in the proliferation and apoptosis of ovarian epithelial carcinoma cells. MethodsA short-hairpin RNA specifically targeting miR-21 plasmid was constructed, and the recombinant was identified by restriction endonuclease analysis and DNA sequencing. Three experimental groups were included, transfection group (transfected with pSIREN-miR-21 ), negative control group ( transfected with pSIREN-miR-21-neg) and blank control group (without transfection plasmid ). The expression of miR-21 was detected by stem-loop real-time reverse transcription (RT)-PCR in OVCAR3 cells ,and western blot was used to detect the expression of programmed cell death 4 ( PDCD4 ) protein. Tethyl thiazolyl tetrazolium (MTT) and flow cytometry method were used respectively. ResultsRecombinant plasmid (pSIREN-miR-21) was constructed successfully and identified by restriction endonuclease analysis and DNA sequencing. The relative expression level of miR-21 in cells transfection, negative control and blank control group was 0.26 ± 0.08, 1.26 ± 0.21and 1.00 respectively. The level of miR-21 in the cells in transfection group was significantly lower than those in the negtive control and blank control group(P ＜0. 01 ). The gray scale of PDCD4. protein was 1443 ±33,858 ± 19 and 846 ± 16 in the transfection group, negative control and blank control group respectively. The value of PDCD4 in transfection group was higher than other control groups, and there were significantly difference among them( P ＜0. 01 ). Moreover, the optical density of the cells in transfection group was 0. 661 ±0. 015,significantly lower than those in two control groups (0. 848 ± 0. 150 for negative control, 0. 935 ± 0. 133 for blank control,P ＜ 0. 01 ). Forty-eight hours after tranfection, the rate of viable apoptotic cell was significantly higher than negative control and blank control group [(25.821 ± 0. 763 )％ vs.(0. 010 ± 0. 003 ) ％ vs. (0. 238 ± 0. 023) ％ ; P ＜ 0. 01];72 hours after tranfection, the rates of viable apoptotic cell and necrotic cell were all higher than the two control groups [the rate of viable apoptotic cell was ( 30. 480 ±0. 821 ) ％, ( 7. 792 ± 0. 312 ) ％ and ( 7. 033 ± 0. 257 ) ％ respectively ( P ＜ 0. 01 ) ; the rate of necrotic cell was (3.558 ±0.211)％, (1.557 ±0.067)％ and (1.049 ±0.028)％, respectively (P＜0. 01)].ConclusionmiR-21 might play an important role in the proliferation and apoptosis of ovarian epithelial carcinoma cells through negatively control the expression of PDCD4.