Objective To investigate the expression of microRNA-29b (miR-29b) in gastric cancer cell line GC9811 and high peritoneal metastatic gastric cancer cell line GC9811-P and its effect on invasion,proliferation and apoptosis.Methods The relative quantitative expression of miR-29b was detected by quantitative realtime polymerase chain reaction(qRT PCR).GC9811 cells were divided into three groups,miRNA down-regulated and transfected with lentiviruses LV-miR-29b inhibitor group,negative control group with negative transfection,and untransfected blank control group.GC9811-P cells were divided into three groups,miR-29b up regulated and transfected with lentiviruses LV miR 29b group,negative control with negative transfection group,and untransfected blank control group.The cell invasion ability was detected with Transwell assay,the cell proliferation ability was measured by methyl-thiazolyl tetrazolium (MTT) test,the colony forming ability was determined by plate colony formation assay,and the apoptosis was tested by flow cytometry.The expressions of miR 29b and secreted protein,acidic and rich in cystenie (SPARC) in gastric cell line GC9811-P,GC9811,MKN28M,MKN28NM and normal gastric cell line GES were determined by qRT-PCR,and the correlation was analyzed.Two independent samples t test or SNK-q test was performed for mean comparison between two groups,and one way analysis of variance was used for mean comparison among three groups.Results The relative quantitative expression of miR-29b inGC9811-P (0.21±-0.04) was significantly lower than that of GC9811 (1.00±0.03,t 28.140,P＜ 0.01).After GC9811 cells transfected with lentiviruses LV-miR-29b inhibitor,the expression of miR-29b (0.21±0.04) was significantly lower than that of control group (0.89±0.07) and blank control group (1.00±0.04,q 12.76,14.73,both P＜0.01).Compared with negative group,the transmembrane cell number and the clonality of miRNA up-regulated group raised(274.33± 9.03 vs 110.67 ± 13.69,t=9.981,P＜0.01;131.33±4.91 vs69.67±2.33,t 11.340,P＜0.01),and the results of MTT test also shows the proliferation was increased.After GC9811-P cells transfected with LV-miR-29b,the expression of miR 29b (4.08±0.20) was significantly higher than that of negative control group (1.15±0.05) and blank control group (1.00±0.10,q=21.73,22.81,both P＜0.01).Compared with negative group,the transmembrane cell number and the clonality of miRNA up regulated group reduced (51.33±5.55 vs 104.00±6.24,t 6.305,P＜0.01; 48.00±5.51 vs 113.33±5.17,t 11.340,P＜0.01),and the results of MTT test also shows the proliferation was weakened.Apoptosis assays demonstrated miR-29b promoted apoptosis; however,the difference was not statistically significant.The expression of miR-29b was negatively correlated with SPARC mRNA in gastric cancer cells (r=-0.97,P=0.03).Conclusions The low expression of miR-29b in high peritoneal metastatic gastric cancer cell inhibited the ability of invasion and proliferation.MiR-29b might be a new target of inhibiting peritoneal metastasis in gastric cancer.

Although ethylene glycol (EG) has been widely used for embryo cryopreservation in domestic animals, few attempts were made to use this molecule to freeze mouse and human embryos. In the few studies that used EG for slow-freezing of mouse and human embryos, complicated protocols for human embryos were used, and the protocols need to be simplified. Besides, freezing mouse morula with EG as a cryoprotectant has not been reported. In this paper, we studied the effects of embryo stages, EG concentration, duration and procedure of equilibration, sucrose supplementation and EG removal after thawing on the development of thawed mouse embryos, using the simple freezing and thawing procedures for bovine embryos. The blastulation and hatching rates (81.92% +/- 2.24% and 68.56% +/- 2.43%, respectively) of the thawed late compact morulae were significantly (P < 0.05) higher than those of embryos frozen-thawed at other stages. When mouse late compact morulae were frozen with different concentrations of EG, the highest rates of blastocyst formation and hatching were obtained with 1.8mol/L EG. The blastulation rate was significantly higher when late morulae were equilibrated in 1.8 mol/L EG for 10 min prior to freezing than when they were equilibrated for 30 min, and the hatching rate of embryos exposed to EG for 10 min was significantly higher than that of embryos exposed for 20 and 30 min. Both rates of blastocyst formation and hatching obtained with two-step equilibration were higher (P < 0.05) than with one-step equilibration in 1.8 mol/L EG. Addition of sucrose to the EG-based solution had no beneficial effects. On the contrary, an increased sucrose level (0.4 mol/L) in the solution impaired the development of the frozen-thawed embryos. In contrast, addition of 0.1 mol/L sucrose to the propylene glycol (PG)-based solution significantly improved the development of the frozen-thawed embryos. Elimination of the cryoprotectant after thawing did not improve the development of the thawed embryos. The cell numbers were less (P < 0.05) in blastocysts developed from the thawed morulae than in the in vivo derived ones. In summary, embryo stage, EG concentration, duration and procedure of equilibration and sucrose supplementation had marked effects on development of the thawed mouse embryos, and a protocol for cryopreservation of mouse embryos is recommended in which the late morulae are frozen in 1.8 mol/L EG using the simple freezing and thawing procedures of bovine embryos after a two-step equilibration and the embryos can be cultured or transferred without EG removal after thawing.
Animals ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dose-Response Relationship, Drug ; Embryo, Mammalian ; drug effects ; physiology ; Embryonic Development ; physiology ; Ethylene Glycol ; pharmacology ; Female ; Mice ; Morula ; physiology ; Pregnancy ; Sucrose ; pharmacology

Objective:To explore the association between atypical small acinar proliferation(ASAP)and subsequent diagnosis of intermediate and high risk prostate cancer(PCa),and analyze whether delaying repeat biopsy timing is safe and effective.Meth-ods:From June 2000 to June 2022,we retrospectively analyzed the clinical data of 276 patients accepting prostatic biopsy and diag-nosed with ASAP in China-Japan Union Hospital of Jilin University.54.7％(151/276)patients had a repeat biopsy.We used statis-tic methods to process the data.Results:25.2％(38/151)patients were diagnosed with PCa on repeat biopsy.Among them,78.9％(30/38)patients had Gleason score(GS)3+3 and 21.1％(8/38)had GS 3+4 disease.There were 4 and 6 patients got RP re-spectively in the two cohorts.Only 5.3％(8/151)of ASAP patients were diagnosed as intermediate risk PCa in repeated biopsy and specially,no high risk PCa was identified in our study.Conclusion:It was safe and valid to delay the repeat biopsy.

BACKGROUNDData on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.

METHODSThe survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.

RESULTSThe analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).

CONCLUSIONSThe prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.

Adult ; Aged ; Awareness ; Female ; Humans ; Hypertension ; complications ; epidemiology ; therapy ; Male ; Middle Aged ; Prevalence ; Renal Insufficiency, Chronic ; complications