Optimized study of an in vitro 3D culture of preantral follicles in mice

Background@#in vitro culture of preantral follicles is a promising technology for fertility preservation. @*Objectives@#This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. @*Methods@#The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E 2 ) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence.The ability of oocytes to fertilize was assessed using in vitro fertilization. @*Results@#The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 (p < 0.05). Meanwhile, the E 2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 (p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro. @*Conclusions@#The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBSfree system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

rBMSCs/ITGA5B1 Promotes Human Vascular Smooth Muscle Cell Differentiation via Enhancing Nitric Oxide Production

BACKGROUND AND OBJECTIVES: Previous studies have shown that integrins alpha5beta1 (ITGA5B1) gene-modified rat bone marrow mesenchymal stem cells (rBMSCs) could prevent cell anoikis and increase the nitric oxide (NO) production. Here we examined the capability of rBMSCs/ITGA5B1 on the phenotype modulation of Human Pulmonary Artery Smooth Muscle Cell (HPASMC) in vitro. METHODS AND RESULTS: The synthetic (dedifferentiated) phenotype of HPASMC was induced by monocrotaline (MCT, 1μM) for 24 h and then co-cultured with rBMSCs/ITGA5B1 in a transwell culture system. The activation of NO/cGMP (nitric oxide/Guanosine-3′, 5′-cyclic monophosphate) signaling was investigated in HPASMC. The changes of pro-inflammatory factors, oxidative stress, vasodilator, vasoconstrictor, contractile and synthetic genes, and the morphological changes of HPASMC were investigated. The results of this study showed that the NO/cGMP signal, endothelial nitric oxide synthase (eNOS) expression, the expression of the vasoprotective genes heme oxygenase-1 (HMOX1) and prostaglandin-endoperoxide synthase 2 (PTGS2) were increased, but the expression of transforming growth factor-β1 (TGF-β1), CCAAT/enhancer-binding proteins delta (Cebpd), Krüppel-like factor 4 (KLF4), and activating transcription factor 4 (ATF4) were reduced in MCT treated HPASMC co-cultured with rBMSCs/ITGA5B1. The synthetic smooth muscle cells (SMCs) phenotype markers thrombospondin-1, epiregulin and the vasoconstrictor endothelin (ET)-1, thromboxane A2 receptor (TbxA2R) were down-regulated, whereas the contractile SMCs phenotype marker transgelin expression was up-regulated by rBMSCs/ITGA5B1. Furthermore, rBMSCs/ITGA5B1 promoted the morphological restoration from synthetic (dedifferentiation) to contractile (differentiation) phenotype in MCT treated HPASMC. CONCLUSIONS: rBMSCs/ITGA5B1 could inhibit inflammation and oxidative stress related genes to promote the HPASMC cell differentiation by activation NO/cGMP signal.

Advances in haplotype analysis technique / 生物工程学报

Haplotype is the combination of a series of genetic mutations that coexist on a single chromosome, each of which has its own unique haplotypes. As a common data analysis method, the analysis of haplotype is effective for the localization of heterozygosis SNPs on single chromosome, the excavation of disease genes and the search of maladies treatments. It mainly includes indirect computational inferential method and direct experimental method. In this review we introduced various haplotype analysis methods and applications, especially two important ones: single-molecule dilution and contiguity-preserving transposition sequencing common technology. Meanwhile, further research prospects on haplotype sequencing were proposed.

Study on hemostatic effect of different methods of oxytocin in laparoscopic myomectomy / 中国综合临床

Objective To examine the hemostatic effect of oxytocin in laparoscopic myomectomy,and to clarify the optimal dosage of oxytocin in laparoscopic myomectomy. Methods From January 2016 to July 2017, ninety patients with myoma of uterus who underwent laparoscopic myomectomy in Tangshan Union Medical College Hospital were divided into two groups: group A and group B,45 cases in each group. In group A,the patients were given 20U oxytocin and 5％ glucose injection 250 ml via intravenous drip, and in group B, the patients were injected with 20U oxytocin on the muscular wall of uterus myoma during operation. The preoperative, postoperative basic situation, intraoperative blood loss, operation time, intraoperative and postoperative adverse reactions and the time of hospitalization were compared between the two groups. Results The average amount of blood loss during operation in group B was (76. 5±15. 8) ml,significantly lower than that in group A((99. 8±20. 7) ml),and the operation time in group B was (71. 2±10. 7) min,shorter than that in group A ((90. 1±18. 4) min) The difference was statistically significant (t=1. 788、2. 878,P<0. 05). At 5 minutes,30 minutes and 180 minutes after medication, the systolic blood pressure of patients in group A was (132. 2±1. 4) mmHg,(120. 7±5. 3) mmHg,(110. 9±1. 8) mmHg and the diastolic blood pressure was (80. 9 ±12. 5) mmHg,(71. 4±12. 6)mmHg,(70. 1±2. 8) mmHg,while the systolic blood pressure in the group B was (140. 4±11. 1) mmHg,(131. 2±4. 5) mmHg,(121. 4±11. 2) mmHg and the diastolic blood pressure was (93. 4±8. 1) mmHg,(86. 4±7. 1) mmHg,(81. 1±1. 5) mmHg,the differences were statistically significant (systolic blood pressure:Ftime=68. 57,P<0. 001;FgrouP=16. 54,P<0. 001;Finteraction=4. 34,P=0. 011;diastolic blood pressure:Ftime=45. 16,P<0. 001;FgrouP=10. 79,P=0. 024;F interaction=4. 16,P=0. 037);at the same treatment time,the systolic blood pressure and diastolic blood pressure in group B were higher than those in group A,the differences were statistically significant (P<0.05).Conclusion Laparoscopic uterine myomectomy with oxytocin hemostasis, intrauterine myoma wall injection method can significantly reduce intraoperative blood loss and shorten the operation time,the effect is obviously better than intravenous drip,and it is conducive to the operation.

Study on hemostatic effect of different administration methods of hemabate in laparoscopic myomectomy / 中国综合临床

Objective To investigate the hemostatic effect of different administration Methods of hemabate in laparoscopic myomectomy, and to clarify the best mode of hemabate in laparoscopic myomectomy.Methods Ninety patients were randomly divided into two groups,group A and group B,45 patients in each group,group A was given intramuscular injection of hemabate 0.25 mg 30 minutes before operation,and in group B,the patients were injected with hemabate 0.25 mg at the muscular wall of uterine leiomyoma during operation.The preoperative,postoperative basic situation,intraoperative blood loss,operation time,intraoperative and postoperative adverse reactions and length of hospital stay were compared between the two groups.Results The average amount of blood loss during operation in the group B was(67.6±17.6)ml, significantly less than that in the group A((88.7±21.6)ml),the operation time in the group B was(68.1 ±18.1)min,shorter than that in the group A(83.1 ± 17.5)min(t=1.877,2.787,P<0.05).The systolic blood pressure of patients in the group A after 15,30,45 and 180 minutes of medication were(155.6±10.31) mmHg,(141.2±5.7)mmHg,(138.6±10.3)mmHg,(127.5±7.4)mmHg,respectively and the diastolic blood pressure were(90.5±8.8)mmHg,(85.5±8.0)mmHg,(80.6±6.3)mmHg,(72.2±7.1)mmHg,while the systolic blood pressure were(141.1±8.3)mmHg,(120.5±6.1)mmHg,(125.7±4.6)mmHg,(118.8±5.5) mmHg and diastolic blood pressure were(81.2 ± 11.4)mmHg,(70.4 ± 10.1)mmHg,(71.5 ± 5.8)mmHg, (66.5±1.2)mmHg in the group B,the differences were statistically significant(systolic blood pressure,Ftime=79.75,P<0.001;FgrouP=25.45,P<0.001; Finteraction=5.48,P=0.041; diastolic blood pressure,Ftime=46.61,P<0.001;FgrouP=11.97,P=0.021,Finteraction=4.05,P=0.034).Conclusion The application of hemabate for hemostasis in laparoscopic myomectomy surgery and uterine fibroids at the muscle wall injection medication can significantly reduce the amount of blood loss during operation,shorten the operation time,and at the same time,the effect on blood pressure was lower than that by preoperative hemabate,is beneficial to the development of operation,with obvious advantages.

Study on hemostatic effect of different administration methods of hemabate in laparoscopic myomectomy / 中国综合临床

Objective To investigate the hemostatic effect of different administration Methods of hemabate in laparoscopic myomectomy, and to clarify the best mode of hemabate in laparoscopic myomectomy.Methods Ninety patients were randomly divided into two groups,group A and group B,45 patients in each group,group A was given intramuscular injection of hemabate 0.25 mg 30 minutes before operation,and in group B,the patients were injected with hemabate 0.25 mg at the muscular wall of uterine leiomyoma during operation.The preoperative,postoperative basic situation,intraoperative blood loss,operation time,intraoperative and postoperative adverse reactions and length of hospital stay were compared between the two groups.Results The average amount of blood loss during operation in the group B was(67.6±17.6)ml, significantly less than that in the group A((88.7±21.6)ml),the operation time in the group B was(68.1 ±18.1)min,shorter than that in the group A(83.1 ± 17.5)min(t=1.877,2.787,P<0.05).The systolic blood pressure of patients in the group A after 15,30,45 and 180 minutes of medication were(155.6±10.31) mmHg,(141.2±5.7)mmHg,(138.6±10.3)mmHg,(127.5±7.4)mmHg,respectively and the diastolic blood pressure were(90.5±8.8)mmHg,(85.5±8.0)mmHg,(80.6±6.3)mmHg,(72.2±7.1)mmHg,while the systolic blood pressure were(141.1±8.3)mmHg,(120.5±6.1)mmHg,(125.7±4.6)mmHg,(118.8±5.5) mmHg and diastolic blood pressure were(81.2 ± 11.4)mmHg,(70.4 ± 10.1)mmHg,(71.5 ± 5.8)mmHg, (66.5±1.2)mmHg in the group B,the differences were statistically significant(systolic blood pressure,Ftime=79.75,P<0.001;FgrouP=25.45,P<0.001; Finteraction=5.48,P=0.041; diastolic blood pressure,Ftime=46.61,P<0.001;FgrouP=11.97,P=0.021,Finteraction=4.05,P=0.034).Conclusion The application of hemabate for hemostasis in laparoscopic myomectomy surgery and uterine fibroids at the muscle wall injection medication can significantly reduce the amount of blood loss during operation,shorten the operation time,and at the same time,the effect on blood pressure was lower than that by preoperative hemabate,is beneficial to the development of operation,with obvious advantages.

Detection of vitamin D in sera of patients with atopic dermatitis and its clinical significance / 中华皮肤科杂志

Objective To measure the serum levels of vitamin D,total immunoglobulin E (tIgE),interleukin-4 (IL-4) and IL-6 in patients with atopic dermatitis (AD),to evaluate the association between vitamin D and severity of AD,and to investigate the role of vitamin D in inflammatory and immunoregulatory processes during the occurrence of AD.Methods Peripheral blood samples were collected from 37 patients with AD (AD group) and 30 healthy controls (control group).The serum levels of vitamin D,tIgE,and IL-6 were measured by chemiluminescent sandwich enzyme immunoassay,and those of IL-4 by enzyme-linked immunosorbent assay.The severity of AD was assessed by the SCORing atopic dermatitis (SCORAD) score.The t test or Mann-Whitney U test was performed to assess the differences in vitamin D,tIgE,IL-4 and IL-6 levels between the AD group and control group,chi-square test to compare the proportion of patients with vitamin D deficiency,insufficiency and sufficiency,and Pearson's correlation analysis or Spearman's correlation analysis to evaluate the correlations between the SCORAD score and serum levels of vitamin D,tIgE,IL-4 and IL-6.Results Compared with the control group,the AD group showed significantly decreased serum levels of vitamin D (24.77 ± 9.29 vs.28.98 ± 6.87 μg/L,t =2.015,P =0.048),but significantly increased serum levels of tIgE (137.68 [37.59-414.53] vs.45.16 [14.56-112.12] IU/ml,Z =-3.399,P =0.001),IL-4 (8.86 ± 4.83 vs.4.78 ± 3.07 ng/L,t =4.147,P ＜ 0.001) and IL-6 (6.53 [3.99-15.30] vs.4.58[2.85-8.17] ng/L,Z =-2.173,P =0.030).Among patients with AD,the SCORAD score was negatively correlated with serum levels of vitamin D (r =-0.505,P =0.001),positively correlated with those of tIgE (r =0.531,P =0.001) and IL-4 (r =0.519,P =0.001),but uncorrelated with those of IL-6 (r =-0.139,P =0.411).There were significant differences in the proportion of patients with vitamin D deficiency,insufficiency and sufficiency between the AD group and control group (x2 =8.762,P =0.013).AD patients with vitamin D deficiency showed significantly increased serum levels of tIgE (2846.87 [319.02-7300.00] IU/ml) and IL-4 ([16.37-2.05] ng/L) compared with those with vitamin D insufficiency (110.07 [26.20-501.48] IU/ml,P =0.045;[8.28 ± 4.48] ng/L,P =0.011) and those with vitamin D sufficiency (123.93 [91.61-273.68] IU/ml,P =0.024;[8.00 ± 4.63] ng/L,P =0.041).In addition,serum levels of IL-6 were also higher in patients with vitamin D deficiency than in those with vitamin D sufficiency (15.10 [8.49-30.72] vs.6.22 [4.47-9.47] ng/L,P =0.011].Conclusions Vitamin D deficiency or insufficiency exists in patients with AD.Vitamin D deficiency is correlated with high serum levels of tIgE,IL-4 and IL-6,and the severity of AD is closely correlated with increased serum levels of tIgE and IL-6 as well as decreased serum levels of vitamin D.

Effect of remifentanil pretreatment on myoclonus after etomidate injection / 中国医师进修杂志

Objective To evaluate the effect of remifentanil pretreatment on myoclonus after etomidate injection.Methods Sixty patients undergoing scheduled for intestinal endoscopy under general anesthesia were divided into remifentanil group and control group by random digits table method with 30 cases each.Before 0.3 mg/kg etomidate was given,the patients in remifentanil group and control group were pretreated with remifentanil 0.5 μ g/kg or amount of 0.9％ sodium chloride.Myoclonus was observed and recorded.Nausea,pruritis and apnea were observed after procedures.Results The incidence of myoclonus was 73.3％(22/30) in control group,mild in 11 cases,moderate in 10 cases and severe in 1 case.The incidence of myoclonus was 6.7％(2/30) in remifentanil group and mild in 2 cases.There was significant difference in the incidence of myoclonus between two groups (P ＜ 0.01).In control group,the ratio of myoclonus was 14/16 in males and 8/14 in females.In remifentanil group,male and female each had 1 case of myoclonus.There was no nausea,pruritis and apnea in remifentanil group.Conclusion Pretreatment with 0.5 μ g/kg remifentanil reduces myoclonus after etomidate injection,male patients are more associated with the incidence of myoclonus than female patients.

Effect of ropivacaine combined with dexamethasone sprayed intratracheally in patients with resistant tubes and postoperative sore throat / 中国医师进修杂志

Objective To investigate the effect of ropivacaine combined with dexanmethasone sprayed intratracheally in patients with resistant tubes and postoperative sore throat.Methods Eighty patients ASA Ⅰ-Ⅱ grade undergoing laparoscopic cholecystectomy were randomly divided into four groups by random digits tabel method,20 patients in each.2 ml of 0.75％ ropivacaine combined with 1 ml dexamethasone (group A) ; 2 ml of 0.75％ ropivacaine combined with 1 ml 0.9％ sodium (group B) ; 3 ml of 1％ tetracaine (group C); 3 ml of 0.9％ sodium (group D) were immediately intratracheal sprayed before intubation.The resistant tubes and degree of postoperative 24 h sore throat [by visual analog scale (VAS) scores],comfort [by Bruggrmann comfort scale (BCS) scores] and the incidence of sore throat were recorded.Results The resistant tubes appraised rate in group A,B and C was higher than that in group D [65.0％ (13/20),60.0％ (12/20),70.0％ (14/20) vs.5.0％ (1/20)] (P ＜ 0.05),but there was no significant difference among group A,B and C (P＞ 0.05).The incidence of sore throat at 24 h after surgery in group A,B and C was lower than that in group D [20.0％(4/20),25.0％ (5/20),25.0％ (5/20) vs.90.0％ (18/20)] (P ＜ 0.05),but there was no significant difference among group A,B and C (P＞ 0.05).The VAS and BCS scores in group A were better than those in group B,C and D [(0.52 ± 1.14) scores vs.(1.68 ± 1.42),(1.59 ± 1.33),(4.22 ± 1.95) scores,(2.80 ± 1.54) scores vs.(1.60 ± 1.19),(1.80 ± 1.20) and (0.45 ± 0.81) scores] (P ＜ 0.05),but there were no significant differences between group B and group C (P ＞ 0.05).Conclusion Ropivacaine combined with dexamethasone sprayedintratracheally can significantly increase the degree of postoperative resistant tubes,decrease the rate of sore throat after general anesthesia,and increase patient' s comfort.