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

Objective:To explore the risk factors of recurrent contralateral hip fracture within 2 years after operation in elderly patients with hip fracture, construct a nomogram prediction model and validate the model.Methods:A total of 601 elderly patients with hip fracture who underwent surgical treatment in Department of Orthopedics in First Hospital of Shanxi Medical University from June 2018 to June 2020 were selected as research objects by the convenient sampling method. They were divided into the modeling set ( n=421) and the verification set ( n=180). According to the incidence of recurrent contralateral hip fracture within 2 years of follow-up, the modeling set was divided into the recurrent fracture group and the normal group, and the clinical data of the two groups were compared. Logistic regression was used to analyze the risk factors for recurrent contralateral hip fracture in elderly patients within 2 years after surgery. R 3.6 software was used to build a risk factor nomogram model for recurrent contralateral hip fracture. Receiver operating characteristic ( ROC) curve and calibration curve were used to evaluate the differentiation and consistency of the model. Results:In 601 elderly patients with hip fracture, the incidence of recurrent contralateral hip fracture within 2 years after surgery was 8.49% (51/601), among which the incidence of modeling set was 8.31% (35/421) and the incidence of verification set was 8.89% (16/180). In the modeling set, the age, female proportion, osteoporosis proportion, combined internal medical disease proportion and malnutrition proportion of patients in the recurrent fracture group were higher than those in the normal group ( P<0.05). Logistic regression analysis showed that gender, age, osteoporosis and combined internal medical diseases were the factors affecting the recurrence of contralateral hip fracture within 2 years after operation in elderly patients with hip fracture ( P<0.05). The equation for constructing a nomogram prediction model was Logit ( P) = -8.521+0.335×age+ 0.116×female +0.341× osteoporosis +0.280 ×combined internal medical diseases. The modeling set predicted the probability of recurrent contralaterial hip fracture according to the nomogram model, and plotted the ROC curve with sensitivity of 0.826, specificity of 0.804, and area under ROC curve ( AUC) of 0.876. The sensitivity of ROC curve of the validation set was 0.788, the specificity was 0.781, and the AUC was 0.830. After internal verification by Bootstrap method, the prediction model of the modeling set and the verification set were well distinguished, and the prediction probability and the actual incidence were well consistent (Hosmer-Lemeshow χ 2=0.462, P=0.674) . Conclusions:Advanced age, female, osteoporosis and combined internal medical diseases are independent risk factors for recurrent contralateral hip fractures in elderly patients with hip fractures within 2 years after surgery. The nomogram model constructed based on this has high predictive efficacy for recurrent hip fractures, which can be used to assess the risk of recurrent fractures and improve the prognosis of patients.

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 observe the auxiliary analgesic effect of electroacupuncture at Hegu and Neiguan combined with in-domethacin suppository in patients undergoing transvaginal ultrasound-guided oocyte retrieval(TUGOR and its effect on the outcome of in vitro fertilization(IVF.METHODS 64 IVF-ET patients undergoing TUGOR were randomly divided into a treatment group and a control group,32 cases in each group.One case dropped out of the treatment group during the treatment.The control group was given indomethacin suppository rectal administration,and the treatment group was given electroacupuncture at Hegu and Neiguan in addition to the treatment of the control group.The patients'tenderness threshold,VAS score,pain grade score,respiratory rate,and pulse rate were evaluated before and after TUGOR operation.The number of oocytes obtained,the rate of two pronuclei(2PN,embryo utiliza-tion rate,and high-quality embryo rate were evaluated after TUGOR operation.The adverse reactions of the two groups were monitored during and after operation.RESULTS After TUGOR operation,the VAS score and pain grade of the treatment group were signifi-cantly lower than those of the control group(P<0.01;the tenderness threshold of the two groups was significantly reduced after opera-tion(P<0.05,P<0.01,and the treatment group was better than the control group(P<0.05;the incidence of nausea during opera-tion,abdominal distension and nausea 48 h after operation in the treatment group were better than those in the control group(P<0.05;the high-quality embryo rate in the treatment group was better than that in the control group(P<0.05.CONCLUSION Electroacupuncture at Hegu and Neiguan combined with indomethacin suppository can effectively assist in analgesia,and can reduce the incidence of adverse reactions during and after TUGOR to varying degrees,and may have certain advantages in improving the rate of high-quality embryos.

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.