1.Diversity analysis of luminal and mucosa-associated microbial communities in obese mice
Qianyun MAO ; Yun SUN ; Chen WANG
Chinese Journal of Internal Medicine 2018;57(10):743-748
Objective Gut microbiota have been reported to be able to regulate host metabolism and is closely associated to obesity. The purpose of this study was to explore the differences between the diversity of luminal and mucosa-associated microbial communities in obese mice. Methods Colonic luminal contents and colonic mucosa were separately collected from 10 obese mice fed with high-fat diet for 12 weeks. DNA of gut microbiota was extracted and micro flora populations were analyzed by Illumina sequencing. Species annotation, diversity analysis, and species difference analysis were conducted. Results The microbial flora from colonic contents had similar richness, evenness and overall structure to those from colonic mucosa (ACE index 250 vs. 285, Chao index 257 vs. 291, Shannon index 3.84 vs. 3.97, Simpson index 0.05 vs. 0.06,all P>0.05). However, there were differences in the microbial composition on specific levels. At the phylum level, colonic contents had higher abundance of Bacteroidetes (56.08% vs. 27.25%, P=3.21×10-5), while colonic mucosa had higher abundance of Firmicutes (49.09%vs. 34.27%, P=0.03) and proteobacteria (18.48% vs. 3.62%, P=0.0009). At the genus level, butyrate-producing bacteria-Lactobacillus was more abundant in colonic content (LDA score=3.89), whereas gram-negative genus Helicobacter, Sphingomonas and Desulfovibrio were relatively abundant in colonic mucosa (LDA score=4.78, 3.59 and 4.11, respectively). Conclusion There were differences in microbial composition at the phylum and genus levels between microbial flora from colonic contents and colonic mucosa, although they had similar richness, evenness and overall structure.
2.Fertilization of in vitro matured human oocytes by intracytoplasmic sperm injection (ICSI) using ejaculated and testicular spermatozoa.
Yun QIAN ; Ting FENG ; Juan CHEN ; Ling-Bo CAI ; Jia-Yin LIU ; Yun-Dong MAO ; Wei DING ; Jia-Hao SHA
Asian Journal of Andrology 2005;7(1):39-43
AIMTo evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI).
METHODSFifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration.
RESULTSA total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term.
CONCLUSIONIt appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.
Adult ; Cell Culture Techniques ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; therapy ; Infertility, Male ; therapy ; Male ; Oocytes ; growth & development ; Pregnancy ; Pregnancy Rate ; Semen ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Testis ; cytology

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