Zichong granules promote differentiation of ovarian granulosa-like cells from human embryonic stem cells in vitro.

OBJECTIVE: To investigate whether Zichong granules (, ZCKL), a very effective herbal formula for treating infertility, have an impact on the differentiation of ovarian granulosa cells from human embryonic stem cells (hESCs) in vitro, and to explore the cellular mechanisms of its clinical effects. METHODS: Serum from ZCKL-medicated rats was prepared and used to treat mesoderm cells derived from hESCs for 6 d. Normal rat serum and a set of growth factors were used as negative and positive controls, respectively. RESULTS: ZCKL-medicated rat serum, but not normal rat serum, induced hESCs-derived mesoderm cells to differentiate into functional ovarian granulosa-like cells (OGLCs) in a similar manner to defined growth factors. The induced OGLCs resembled the morphology of native human granulosa cells, expressed granulosa cell-specific markers at both the mRNA and protein levels, produced high levels of estradiol and strongly responded to follicle-stimulating hormone stimulation. Furthermore, mRNA levels of follistatin, mothers against decapentaplegic homolog 8 and bone morphogenetic protein 6 were dynamically changed during the process. CONCLUSION: In the ZCKL treatment of infertility, one mechanism by which ZCKL may act is by influencing ovarian granulosa cell differentiation and development, possibly through the follistatin and BMP/SMAD signaling pathways.

Clinical significance of plasma anti-TOPO48 autoantibody and blood survivin-expressing circulating cancer cells in patients with early stage endometrial carcinoma.

PURPOSE: To examine the clinical significance of an autoantibody (AAb) against a novel tumor-associated antigen (TAA) derived from human DNA-topoisomerase I, termed as TOPO48 AAb, and peripheral blood survivin-expressing circulating cells (CCC) in patients with early stage endometrial cancer (EC). METHODS: Blood samples were collected from 80 patients with early stage EC and 80 age-matched healthy subjects. Plasma levels of the TOPO48 AAb were measured with a specific antibody capture enzyme-linked immunosorbent assay (ELISA) and blood survivin-expressing CCC assessed with a reverse transcription-polymerase chain reaction products based on a hybridization-enzyme-linked immunosorbent assay (RT-PCR-ELISA). Sixty patients were followed up for 36 months after the initial assay test. RESULTS: There were 75% and 60% samples with positive levels of the TOPO48 AAb and survivin-expressing CCC in the cancer patients, respectively. However, the cumulative positive rate of combination of the two markers was increased to 93.3% with 0.927 (95% CI 0.871-0.984) of area under the curve (AUC) in receiver operating characteristic (ROC) curve analysis. During the follow-up period, patients with positive TOPO48 AAb but negative surviving-expressing CCC had a higher survival rate and a longer survival time than those with negative AAb but positive CCC (P = 0.01). CONCLUSIONS: The combination of TOPO48 AAb and survivin-expressing CCC may be used as a novel recipe to improve the efficiency of early diagnosis and provide more accurate prognostic prediction in patients with early stage EC.

A misdiagnosed case of osteoid osteoma of L5.

Clinically, it is difficult to differentiate osteoid osteoma, more than 50% of which occur in the fibia or tibia, from other diseases, i.e. spinal degenerative diseases, inflammatory and noninflammatory arthritis. In this case report, we presented an unusual case of lumbar osteoid osteoma in a 38-year-old male, who experienced low back pain and sciatica as initial symptoms. The patient was initially misdiagnosed as lumbar disc herniation for more than 10 years. With the usage of computed tomography (CT) and magnetic resonance imaging (MRI), the patient was finally diagnosed as osteoid osteoma in L5. To our knowledge, spinal osteoid osteoma with sciatica as initial symptoms has never been reported. Although lumbar vertebra osteoid osteoma is clinically uncommon, it should be taken into consideration especially when patients experience long duration of pain in lumbar.

Common Variants in OPG Confer Risk to Bone Mineral Density Variation and Osteoporosis Fractures.

Although many common variants have been identified for bone mineral density (BMD) and osteoporosis fractures, all the identified risk variants could only explain a small portion of heritability of BMD and osteoporosis fractures. OPG belongs to the tumor necrosis factor receptor superfamily, which plays a crucial role in bone remodeling and is thus a promising candidate gene of osteoporosis. Several studies have explored the association of OPG variants with BMD or osteoporosis fractures, however, the results remain inconsistent among different populations. In the study, we first assessed the relationship between OPG variants and BMD or osteoporosis fractures in our sample size (227 subjects with postmenopausal osteoporosis and 189 controls), and then performed a systematic meta-analysis. Among the nine SNPs genotyped, rs6469804 and rs2073618 showed significant associations with both BMD and osteoporotic fractures, while rs3102735 was only associated with BMD in our samples (P < 0.05). For meta-analyses, data for a total of 12 SNPs were pooled (4725 patients and 37804 controls), and five SNPs, including rs6993813, rs6469804, rs3134070, rs2073618 and rs3102734, showed association with osteoporosis fractures (P < 0.05). On light of the above analysis, we believe that OPG is one promising susceptibility gene of BMD or osteoporotic fractures.

[Effects of Zuogui pill, Yougui pill and relative compositions on differentiation towards germ cells of mouse embryonic stem cell 1B10].

OBJECTIVE: To investigate the effects of Zuogui pill, Yougui pill and the relative compositions on the differentiation towards germ cells of stem cells. METHOD: The rat drug sera for Zuogui pill, Yougui pill and the common composition of Zuogui pill and Yougui pill were prepared respectively as the experimental drugs; the mouse embryonic stem cell 1B10 (MESC-1B10) was used as the representative of stem cells; the above rat drug sera were used to intervene MESC-1B10 and the process was traced by microscopy imaging; after 72 h of the intervention, the RNAs were extracted from the different intervened MESC-1B10, cDNAs were synthesized immediately and finally the Real-time quantitative PCR (qPCR) was used to measure the expression patterns of the 10 reproductive-differentiation-related genes for each intervention. RESULT: The rat drug serum of Zuogui pill (ZGW-RS) significantly up-regulated Oct-4 and SCP3 and significantly down-regulated GDF-9 and Stra8; the rat drug serum of Yougui pill (ZGW-RS) significantly up-regulated Oct-4, GDF-9, Mvh and SCP3 and significantly down-regulated Stra8, Itga6 and Itgb1; the rat drug sera for the common composition of Zuogui pill and Yougui pill (ZGWYGW-RS) significantly up-regulated Oct-4, SCP3 and ZP3 and significantly down-regulated GDF-9, Stra8, Itga6 and Itgb1. CONCLUSION: ZGW-RS can initiate the change towards meiosis, but can not start the reproductive differentiation of MESC-1B10; YGW-RS can initiate the change towards meiosis, and can also start the reproductive differentiation of MESC-1B10 towards female germ cells; ZGWYGW-RS can initiate the change towards meiosis, and can lightly start the reproductive differentiation of MESC-1B10 towards female germ cells but the inductive effect is smaller than YGW-RS. The experimental results, on one hand, strengthen the knowledge about the influence of the relative compositions of Zuogui pill and Yougui pill on the reproductive differentiation of stem cells, on the other hand, help to explain the mechanism of the treatment of the infertility by Zuogui pill and Yougui pill.

Study of oleanolic acid on the estrodiol production and the fat production of mouse preadipocyte 3T3-L1 in vitro.

The women during the menopause period have an increased tendency for the obesity, which represents the more fat production than during the premenopausal period. Although this is not beneficial overall, it could provide a compensatory source for the estrogen production for the menopausal women. So it would be meaningful to find an agent that could inhibit the fat production while does not disturb the total estrogen production by fat tissues. In the present study, the effect of oleanolic acid (OA) on the fat production and the total estrogen production of the differentiating mouse preadipocyte 3T3-L1 as well as the mechanisms behind those effects were preliminarily investigated. The cell line 3T3-L1 was chosen as the model cell because it is usually used for the research about the obesity. During the induced differentiation of 3T3-L1 cells, cells were intervened continuously with OA. The fat production was determined with the oil red staining assay and the total estrogen production was measured with the ELISA assay. Finally, the expression patterns for important genes of the fat production and the estrogen production were studied, respectively with the real-time fluorescence quantitative PCR (qPCR). The results showed that for the differentiating 3T3-L1 cells, OA could significantly inhibit the fat production and did not disturb the total estrogen production significantly. In the mechanism studies, OA was found to significantly down-regulate ACC, the key gene for fat synthesis, which could explain the inhibitory effect of OA on the fat production; OA was also found to significantly up-regulate CYP11A1, CYP17, CYP19, the key genes for the estrogen synthesis and significantly down-regulate CYP1A1, the key gene for the estrogen decomposition, which preliminarily explained the lack of the effect of OA on the total estrogen production. In conclusion, OA was found able to inhibit the fat production while maintaining the total estrogen level and the mechanisms for the above findings were preliminarily clarified, which suggests that OA may be useful to treat the menopausal obesity.

Impacts of You Gui Wan on the expression of estrogen receptors and angiogenic factors in OVX­rat vagina: a possible mechanism for the trophic effect of the formula on OVX­induced vaginal atrophy.

The administration of You Gui Wan (YGW) decoction has been observed to improve vaginal atrophy induced by ovariectomy (OVX) in rats. The aim of the current study was to explore the possible mechanisms underlying this effect. Following OVX, 37 Sprague Dawley female rats were randomly divided into three groups which were orally administered with YGW decoction, saline or estrogen for 11 weeks. In parallel with this, 19 normal and 17 rats with sham-surgery were used as controls. The effects of these treatments on estrogen receptors (ER) and various angiogenic factors, including vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-1 (VEGFR-1), angiopoietin (Ang)1 and 2 and basic fibroblast growth factor (bFGF) in the vagina were compared using immunohistochemistry or quantitative polymerase chain reaction (qPCR). OVX was found to induce significant vaginal atrophy and decrease the expression of ER and various angiogenic factors when compared with the normal and sham-surgery animals (all P<0.05). Estrogen replacement and the administration of YGW decoction reversed the vaginal atrophic process. The hormonal replacement and YGW treatment recovered the protein expression of ER-α and -ß, VEGF and VEGFR-1 and the mRNA levels of ER-α, VEGF, VEGFR-1, Ang1 and 2, and bFGF when compared with OVX-rats with saline, normal and sham-surgery treatments (all P<0.05). Thus, it may be concluded that a possible mechanism underlying the effect of YGW on OVX-induced vaginal atrophy may be the upregulated expression of ER and various angiogenic factors in the vaginal tissue.