Efficacy and Safety of the Chinese Herbal Compound TJAOA101 in Treating Diminished Ovarian Reserve: A Protocol for Multicenter, Prospective, and Pre-Post Study.

OBJECTIVE: Diminished ovarian reserve (DOR) can lead to early menopause, poor fecundity, and an increased risk of disorders such as osteoporosis, cardiovascular disease, and cognitive impairment, seriously affecting the physical and mental health of women. There is still no safe and effective strategy or method to combat DOR. We have developed a novel Chinese herbal formula, Tongji anti-ovarian aging 101 (TJAOA101), to treat DOR. However, its safety and efficacy need to be further validated. METHODS: In this prospective and pre-post clinical trial, 100 eligible patients aged 18-45 diagnosed with DOR will be recruited. All participants receive TJAOA101 twice a day for 3 months. Then, comparisons before and after treatment will be analyzed, and the outcomes, including anti-mullerian hormone (AMH) and follicle-stimulating hormone (FSH) levels and the antral follicle count (AFC), the recovery rate of menopause, and the Kupperman index (KMI), will be assessed at baseline, every month during medication (the intervention period), and 1, 3 months after medication (the follow-up period). Assessments for adverse events will be performed during the intervention and follow-up periods. CONCLUSION: A multicenter, prospective study will be conducted to further confirm the safety and efficacy of TJAOA101 in treating DOR and to provide new therapeutic strategies for improving the quality of life in DOR patients.

[Protective effects of caloric restriction on ovarian function].

OBJECTIVE: To study the protective effects on ovarian function by caloric restriction (CR) and its mechanism. METHODS: Thirty female C57BL/6 mice of 8 weeks old were randomly divided into two groups, including ad libitum (AL) group and caloric restriction (CR) group. The general situation and ovarian function of those mice were compared and evaluated.Ovarian follicles were counted by hematoxylin-eosin staining. Anti-Miillerian Hormone(AMH) mRNA expression of the ovary were detected by using real-time PCR. The concentrations of serum estradiol, progesterone of the mice were measured by ELISA. And the fertility of mice by mating trials were evaluated, SIRT3, Hypoxia inducible factor 1α (HIF-1α) and catalase (CAT) mRNA expression of the mice ovaries were detected by Real-Time PCR. RESULTS: The total follicles were 546 in CR mice and 286 in AL mice. The proportion of primordial follicles were 38.6% (211/546) in ovaries of CR mice and 29.4% (84/286) in ovaries of AL mice, which reached statistical difference. The proportion of atretic follicles 5.3% (29/546) in ovaries of CR mice, compared with 16.8% (48/286) in AL mice, was significantly decreased (P < 0.05). The AMH mRNA expression in CR mice ovaries was 3.37 times of that of AL mice (P < 0.05). The serum concentration of estradiol in CR mice was up to (5.3 ± 1.6) pmol/L, which was much higher than (3.6 ± 1.6) pmol/L in AL mice. While, the progesterone concentration of (0.4 ± 0.3) nmol/L in CR mice was lower than (1.4 ± 0.8) nmol/L in AL mice (P < 0.05).Fertility and survival of offsprings were both improved in CR mice. The expression level of SIRT3 mRNA in CR mice ovary was 1.39 times, CAT was 1.55 times and HIF-1α was 0.31 times of those in AL mice (P < 0.05). CONCLUSIONS: Caloric restriction can delay the ovary aging process through reduce follicle depletion by suppressing follicle recruitment and ovulation. The function of ovarian reserve and reproductive endocrine was effectively protected. Caloric restriction can reduce the incidence of follicular atresia, its mechanism might be associated with anti-oxidative stress.

[Expression and significance of Smad2/3 and HPV16 E7 in cervical intraepithelial neoplasia and cervical carcinoma].

BACKGROUND & OBJECTIVE: Smad proteins are downstream signal proteins of the transforming growth factor-beta (TGF-beta) superfamily, which are intimately related to the genesis of many human carcinomas. Human papillomavirus (HPV) is an important carcinogenic agent of cervical cancer. However, their relationship in the genesis of cervical cancer is unclear. This study was to detect the expression of Smad2/3 and HPV16 E7 protein in different cervical lesions, and to explore their possible roles in tumor genesis and progression. METHODS: The expression of Smad2/3 and HPV16 E7 in 20 specimens of chronic cervicitis, 30 specimens of cervical intraepithelial neoplasia (CIN), and 30 specimens of cervical cancer was detected by SP immunohistochemistry. RESULTS: The positive rates of Smad2/3 were significantly lower in cervicitis and CIN than in cervical cancer (50.0% and 73.3% vs. 93.3%, P<0.05), and significantly lower in cervicitis than in CINIII(P<0.05). The positive rates of HPV16 E7 were 60.0% in cervicitis, 66.7% in CIN, and 83.3% in cervical cancer (P>0.05). The expression of Smad2/3 in cervical cancer had no correlation to clinical stage, pathologic classification, histological grade, and lymph node involvement(P>0.05). Smad2/3 expression was positively correlated to HPV16 E7 infection (r=0.271, P=0.015, R-Spearman). CONCLUSIONS: Overexpression of Smad2/3 may be involved in the genesis of cervical cancer. It is closely correlated to HPV infection in cervical carcinoma.

[Reversal effect of antisense RNA targeting human papillomavirus 16 (HPV16) E6E7 on malignancy of human cervical cancer cell line SiHa].

BACKGROUND & OBJECTIVE: The ubiquitinated degradation of p53 can be induced by human papillomavirus type 16 (HPV16) early gene 6 (E6); the phosphorylated inactivation of pRb can be induced by HPV16 E7. They are closely associated with the carcinogenesis and progression of cervical cancer. This study was to investigate the effects of HPV16 E6E7 antisense RNA on the expression of oncogene E6 and E7 and apoptosis of human cervical cancer cell line SiHa harboring HPV16 genome. METHODS: The antisense sequence of HPV16 E6E7 was cloned into eukaryotic expression vector pEGFP to prepare recombinant plasmid containing E6E7AS, which was transfected into SiHa cells. The mRNA and protein levels of E6 and E7 gene were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Cell proliferation after transfection was evaluated by MTT assay. The apoptosis of transfected SiHa cells was assessed by flow cytometry (FCM) and confocal laser microscopy. RESULTS: After transfection of HPV16 E6E7 antisense RNA, the mRNA and protein levels of HPV16 E6 and E7 in SiHa cells were obviously decreased. The proliferation activity of SiHa/E6E7AS cells was significantly lower than that of SiHa/EGFP and SiHa cells (0.50+/-0.05 vs. 1.01+/-0.06, 1.28+/-0.06, P<0.05). The apoptosis rate was significantly higher in SiHa/E6E7AS cells than in SiHa/EGFP and SiHa cells [(59.3+/-11.3)% vs. (9.4+/-1.8)%, (2.1+/-0.4)%, P<0.05]. The apoptotic cells increased noticeably after transfection. CONCLUSION: Down-regulating HPV16 E6 and E7 with antisense RNA induces apoptosis in SiHa cells, and may be useful for HPV-associated malignancy gene therapy.