ABSTRACT
BACKGROUND: Studies have confirmed that anti-zona zona antibodies can accelerate the destruction and depletion of oocytes, thereby causing premature ovarian failure. OBJECTIVE: To investigate the time of establishing autoimmune premature ovarian failure model in BALB/c mice induced by zona pellucida 3 peptides. METHODS: Thirty healthy female BALB/c mice (7-8 weeks) were randomly divided into immune experimental group (20 mice) and the control group (10 mice). In the experimental group, the mice were given immunization injection starting at 0 week, 0.15 mL of immune reagent injected into the soles at both sides and the lower abdomen. The vaginal exfoliated cell smears were observed every morning to observe the changes in the estrous cycle of the mice. After 2 weeks of injection, 0.15 mL of immune enhancement agent was injected subcutaneously into the same site. On the first day of weeks 4 and 6, the immune agent or immune enhancement agent was injected alternatively. Blood samples were collected before each injection and the serum sex hormone level was measured by enzyme-linked immunosorbent assay. Finally, ovarian tissue and uterus morphology were observed. RESULTS AND CONCLUSION: There was no difference in serum follicle stimulating hormone and estrogen levels between the first immunization injection and 2 weeks after the injection. The serum estrogen level in the experimental group was significantly lower than that in the control group at 4 weeks after injection. The serum estrogen level of the experimental group was significantly lower than that of the control group at 6 weeks after injection (P < 0.05), and meanwhile the serum follicle stimulating hormone level was significantly higher than that of the control group (P < 0.01). The degree of ovarian interstitial fibrosis in the experimental group was more obvious than that in the control group. The ovarian volume decreased and the uterus atrophied in the experimental group. The number of primitive follicles, primary follicles and secondary follicles in mice was significantly lower in the experimental group than the control group, and the number of atresia follicles was significantly higher than the control group (P < 0.05). Therefore, 75 μg of zona pellucida 3 peptides can be used to establish an autoimmune premature ovarian failure disease model in BALB/c mice, and a good modelling effect can be achieved at 6 weeks after immunization.
ABSTRACT
Objective To investigate the regulatory effects of Zishen Yutai Pills on the expression levels of homeboxA10 (HOXA10) and its downstream target gene empty spiracles homebox 2 (EMX2) in the endometria of ovulation-inducing mice at different implantation stages. Methods Seventy-five estrous female Kunming mice were randomly divided into 5 groups, namely normal group, model group 1, model group 2, treatment group 1, treatment group 2, 15 mice in each group. The model group 1 was given short-term protocol for ovulation induction; the model group 2 was given long-term protocol for ovulation induction; the treatment group 1 was given Zishen Yutai pills (at the dose of 0.4 g/mL) on the basis of the protocol for the model group 1; the treatment group 2 was given Zishen Yutai Pills (at the dose of 0.4 g/mL) on the basis of the protocol for the model group 2; the normal group was given intragastric administration or intraperitoneal injection of the same volume of normal saline. The mRNA and protein expression levels of HOXA10 and EMX2 in mouse uterus were detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot method, respectively. Results Compared with the normal group, the mRNA and protein expression levels of HOXA10 were decreased, and the mRNA and protein expression levels of EMX2 were increased in model group 1 and model group 2(P< 0.01). Compared with the corresponding model group 1 and 2, the mRNA and protein expression levels of HOXA10 were significantly up-regulated (P < 0.01) , and the mRNA and protein expression levels of EMX2 were decreased in the treatment group 1 and 2 (P < 0.01), respectively. Conclusion Zishen Yutai Pills may improve mouse endometrial receptivity by up-regulating HOXA10 expression and inhibiting EMX2 expression.
ABSTRACT
Objective To develop multiplex polymerase chain reaction (PCR) combined with reversedotblothy bridization (RDB) method for detection of DNA virus in respiratory samples,and provide a surveillance and rapid diagnosis tool of acute viral respiratory infection.Methods We designed multiple PCR primers and the probes referenced to virus nucleic acid sequences in the National Center for Biotechnology Information (NCBI) database,and fixed specific oligonucleotide probes on the nylon membrane.After multiple PCR amplification of virus DNA of human bocavirus (hBOV),karolinska Institutet (KI),adenovirus (AdV),Washington University polyomaviros (WUPyV),and human parvovirus B19 (HPVBI9),the denaturalized amplification products were hybridized with various specific probes,followed by visualization and analysis of the results.The sensitivity and specificity were tested.At the same time,108 cases of clinical specimens of multiple PCR products were analyzed by reverse spot hybridization detection,and compared to the results of culture method.Results The specific probes of multiple PCR-RDB only hybridized with corresponding amplification products without cross-hybridization reaction with other pathogen.The sensitivity of RDB hybridization was 1 colony-forming units (CFU).The positive rate of 34.26％ (37 cases out of 108 cases) with PCR-RDB method was significandy higher than that 27.78％ (30 cases out of 108 cases) with common test method.Conclusions The multiplex PCR combined with RDB might become a rapid and simple method to detect the DNA virus in respiratory samples,which might be a promising tool for clinical application.