BACKGROUND:The blastula culture medium can assist the development of zygote from the fertilized egg to the blast blastula. The safety and quality of blastula culture medium directly influences the quality of blastula. OBJECTIVE:To evaluate the effect of blastula culture medium on the development of mouse embryos. METHODS: In this study, B6D2F1 mice were used. The female mice were superovulated and mated with male B6D2F1 mice. One day later, the zygotes were colected and cultured in the M16 medium to 4-cel stage. Then, 4-cel stage embryos were transferred into the tested blastula culture medium (experimental group). After 5 days of culture, the forming rate of blastula was examined. Meanwhile, the M16 medium containing endotoxin was used to culture 1-cel mouse zygote (positive control group). The M16 medium with no embryo toxicity was used to culture 1-cel zygote (negative control group). RESULTS AND CONCLUSION:The formation rate of blastula was 0 in the positive group, 87.1% in the negative control group, and 87.3% in the experimental group. From the results, the tested blastula culture medium could assist the 1-cel zygote growing to the stage of blastula, and the formation rate of blastula was above 80%. The tested blastula culture medium had no toxicity to the mouse embryo.

Objective To explore the effect of nicotinaide nucleotide transhydrogenase(NNT) mutation on glucose homeostasis in C57BL/6 mice with mix background. Methods We generated wild type NNT homozygous, mutant NNT homozygous and heterozygous by mating the C57BL/6J (with NNT mutation) and 6N (without NNT mutation). At the age of 4 weeks, those mice were randomly assigned to normal control diet(NCD) or high-fat diet(HFD) for 4 weeks. The body weight was measured every week. At the age of 8 weeks, an intraperitoneal glucose tolerance test(IPGTT) and an intraperitoneal insulin tolerance test (ITT) were performed. Results The body weight growth was not affected by NNT mutation during an HFD fed. NNT mutant mice showed significant glucose intolerance. After 4 weeks of high fat diet, the NNT mutant mice showed a decreased insulin sensitivity, while the glucose excursion curve was not elevated in the heterozygous mice. Conclusion NNT mutation had a significant influence on the phenotype of glucose metabolism and insulin resistance of mice, in particular under a metabolic stress. The phenotypes of heterozygous and homozygous mutant ones differed from each other. When using mice with C57BL/6J and C57BL/6N mixed background in research, NNT mutation should be carefully screened in all metabolic studies.

This study was aimed to detect the expression of miR-665 in the hippocampal of depression rat model treated with Kai-Xin Jie-Yu (KXJY) decoction and bioinformatic analysis of target genes of miR-665,in order to investigate the role of miR-665 in the pathogenesis of depression and the antidepressant mechanism of KXJY decoction.The rat model of chronic stress depression was established and then treated with KXJY decoction for 42 days.The total RNA from hippocampus tissues was extracted.And the relative expression of miR-665 in hippocampus of rats in each group was detected by RT-qPCR.TargetScan and microRNAorg databases were used to predict target genes for miR-665.DAVID database was used to classify GO function and to analyze KEGG signaling pathway of target genes.The results showed that compared with the normal group,the expression level of miR-665 in hippocampus of the model group was significantly higher with significant difference (P ＜ 0.01).Compared with the model group,the expression level of miR-665 in hippocampus of Chinese medicine group and western medicine group decreased significantly with significant difference (P ＜ 0.01).The biological functions of miR-665 target genes were mainly concentrated in the response to organic substance.The signal pathway was mainly concentrated in N-Glycan biosynthesis.It was concluded that miR-665 may be involved in the pathological process of depression,by correcting the abnormal expression of miR-665,which may be one of the antidepressant mechanisms of KXJY decoction.Through the analysis and prediction of the target genes,it provided a certain direction and theoretical basis for further study on the specific mechanism of KXJY decoction intervention on miR-665.

Previous studies have indicated that ERp44 inhibits inositol 1,4,5-trisphosphate (IP(3))-induced Ca(2+) release (IICR) via IP(3)R(1), but the mechanism remains largely unexplored. Using extracellular ATP to induce intracellular calcium transient as an IICR model, Ca(2+) image, pull down assay, and Western blotting experiments were carried out in the present study. We found that extracellular ATP induced calcium transient via IP(3)Rs (IICR) and the IICR were markedly decreased in ERp44 overexpressed Hela cells. The inhibitory effect of C160S/C212S but not C29S/T396A/ΔT(331-377) mutants of ERp44 on IICR were significantly decreased compared with ERp44. However, the binding capacity of ERp44 to L3V domain of IP(3)R(1) (1L3V) was enhanced by ERp44 C160S/C212S mutation. Taken together, these results suggest that the mutants of ERp44, C160/C212, can more tightly bind to IP(3)R(1) but exhibit a weak inhibition of IP(3)R(1) channel activity in Hela cells.
Adenosine Triphosphate ; pharmacology ; Amino Acid Substitution ; Biological Transport ; drug effects ; physiology ; Blotting, Western ; Calcium ; metabolism ; Calcium Signaling ; drug effects ; physiology ; HeLa Cells ; Humans ; Immunoprecipitation ; Inositol 1,4,5-Trisphosphate ; metabolism ; Inositol 1,4,5-Trisphosphate Receptors ; physiology ; Membrane Potentials ; drug effects ; physiology ; Membrane Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Molecular Chaperones ; genetics ; metabolism ; Mutation ; Plasmids ; Transfection

OBJECTIVETo explore a step-by-step exclusive diagnosis and analyze the clinical characters of non-allergic rhinitis (NAR).

METHODSPatients with symptoms (nasal itching, sneezing, rhinorrhea, nasal congestion) were selected to take four-step exclusive diagnosis for NAR and we tried to eliminate the false NAR and retain the true NAR. First step was to exclude the patients who were not suitable for skin prick test (SPT, such as during pregnancy, breastfeeding, asthma, oral antihistamine medication in 7 day, severe skin diseases). The second step was to exclude the patients with positive SPT and the third step was to exclude the patients with 1 level or above of specific sero-immunoglobulin E (sIgE). The fourth step was to exclude the patients with infection rhinitis, clear abnormal nasal structure, drug-induced rhinitis, nasal neoplasm. The remained patients were finally diagnosed as NAR and who were further differential diagnosed as vasomotor rhinitis (VMR) or non-allergic rhinitis with eosinophilia syndrome (NARES) according to the eosinophilia counts in nasal secretion and venous blood. The common characters of patients with NAR were analyzed and their symptoms and quality of life were evaluated by visual analogue scale (VAS) and rhino-conjunctivitis quality of life questionnaire (RQLQ) separately.

RESULTSOne thousand four hundred and thirty-seven patients were included after first step exclusion and 735 cases with negative SPT were remained after second step exclusion. Of 735 patients, 302 were tested in vitro for sIgE and 93 cases with 0 level of sIgE and total IgE were remained after third step exclusion. Sixty-two patients were finally diagnosed as NAR after fourth step exclusion. The NAR diagnosis rate was 51.15% (735/1 437) with negative SPT alone and the NAR diagnosis rate was 29.06% (93/302) with combination of negative SPT and sIgE. Of 62 patients with NAR, 47 patients (75.81%) were diagnosed as VMR and 15 cases (24.19%) as NARES. There were 23 males and 39 females in the 62 patients aged 11 - 77 years. The history was 11-47 months. The biggest numbers of patients with VMR or NARES were among 41-50 years. Their onset ages were among 21-30 years in both two groups. VAS scores of nasal congestion in VMR patients were the highest with significant difference among nasal symptoms (F = 3.958 0, P = 0.009 1). VAS scores of sneezing in NARES patients were the highest but without significant difference among nasal symptoms. There were no difference in seven domain scores of RQLQ and the total mean scores between VMR group and NARES group but the nasal symptoms got the highest scores with significant difference among the seven domains in each group (VMR group, F = 9.771 2, P = 0.000 0;NRAES group, F = 3.226 9, P = 0.006 2).

CONCLUSIONSSPT combined with sIgE may exclude much more patients with AR. Females with NAR are much more than males. Patients with NAR aged 21-30 years. The characters of NAR are helpful to improve our knowledge about NAR. VAS and RQLQ may be a suitable tool in assessment of NAR.

Adolescent ; Adult ; Aged ; Asthma ; diagnosis ; Child ; Diagnosis, Differential ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Quality of Life ; Rhinitis ; diagnosis ; Rhinitis, Allergic ; diagnosis ; Rhinitis, Vasomotor ; diagnosis ; Skin Tests ; Surveys and Questionnaires ; Young Adult

Objective To evaluate the role of 3.0 Tesla magnetic resonance hysterosalpingography (MR?HSG) work?up in the diagnosis of female infertility. Methods Between July 2015 and December 2018, a total of 1 052 infertile women aged from 20 to 40 years in the Affiliated Hospital of Nanjing University of Chinese Medicine were prospectively enrolled in the study. All the patients underwent pelvic plain scanning and X?ray hysterosalpingography (HSG) followed by MR?HSG examination, and the patency of the fallopian tubes as well as the abnormalities of the uterus and ovaries were evaluated. Among which 33 cases were randomly selected. The chi?square test and Kappa test were used to compare the difference and the consistency of the two methods in the evaluation of fallopian tubes. Results MR?HSG and HSG had good consistency in evaluating tubal patency (Kappa=0.88, P<0.01), and there was no statistically significant difference between the two groups (P=0.65). The examination of MR?HSG was successfully completed in 97.1%(1 021/1 052) cases. There were 81.7% (834/1 021) cases had at least one abnormality. Bilateral tubal, uterine and ovarian abnormalitiesoccurred in 42.6% (435/1 021), 34.2% (349/1 021)and 46.8% (478/1 021) cases, respectively. In which tubal abnormalities display the results as follows: bilateral obstructed 4.7% (48/1 021), bilateral poor pass 8.5% (87/1 021), one smooth one obstructed11.7% (119/1 021), one smooth one poor pass 12.6% (129/1 021), and one poor pass one obstructed 5.1% (52/1 021). Conclusion 3.0 T MR?HSG is expected to be a routineexam for evaluating female infertility, which allows a comprehensive assessment of tubal patency and other pelvic abnormalities of infertile women.

Objective To evaluate the efficacy,safety of rigid gas permeable contact lens (RGPCL) wearing for over 5 years in different degrees of keratoconus eyes.Methods A retrospective case study was performed.The clinical data of 217 eyes with different degree of keratoconus from 126 keratoconus patients who fitted with RGPCL in Beijing Tongren Hospital from 2000-2010 over 5 years were analyzed.The eyes were divided into mild keratoconus group (Ks≤45.0 D),moderate keratoconus group (45.0 D＜ Ks ＜ 52.0 D) and severe keratoconus group (Ks ≥52.0 D) according to the severity.Uncorrected visual acuity (UCVA,LogMAR),spectacle corrected visual acuity (SCVA) and RGP corrected visual acuity (RGPVA) was examined before RGPCL wearing and the end of followingup after RGPCL wearing.The refraction,corneal curvature,corneal astigmatism were measured with auto-refractomer/keratometer and keratoconus screening analysis system of computer-assisted corneal topography.Comparisons of the changes of corrected visual acuity and corneal parameters were assessed.Results The RGPVA was 0.09±0.17,0.05±0.07 and 0.07 ±0.07 in the mild,moderate and severe keratoconus group,respectively,showing a significant difference among the three groups (F=0.522,P=0.594);The △Ks was (2.25±5.42),(0.26±3.44) and (-4.52±3.44)D,and △Kf was (2.06±4.98),(1.02±3.41) and (-2.03±5.05)D,and the change value of corneal astigmatism was (0.19±2.87),(-0.78±2.84) and (-2.44±3.77)D in the mild,moderate and severe keratoconus group(all at P＜ 0.05),respectively,with the minimum amount of change in the severe keratoconus group.The variation of differential sector index (△DSI) was-0.33 ± 1.64,0.14±3.01 and-2.11 ±4.28;the variation of center/ surround index (△CSI) was-0.41 ± 1.07,0.03±2.22 and-2.49±4.15;the variation of standard deviation of power (△SDP) was-0.43 ±0.64,-0.02 ±0.89 and-1.67 ± 1.68;the variation of keratoconus prediction index (△KPI)was 0.00±0.07,0.03±0.09 and-0.05±0.11 in the mild,moderate and severe keratoconus group,respectively,and the reduced amount in above parameters was much more in the severe keratoconus group than that in the mild and moderate keratoconus group (all at P＜0.01).Mild conjunctivitis and corneal affection occurred in 12 eyes (5.5％)during the follow-up.Conclusions Long-term wearing RGPCL can improve the visual acuity and slow the tendency of corneal curvature increase in keratoconus eyes,and this procedure is safe and effective for the correction of different degree of keratoconus.