Oestrogen dose tapering during luteal phase does not affect clinical outcomes after hormone replacement treatment-frozen-thawed embryo transfer cycles: a retrospective analysis.

STUDY QUESTION: Does oestrogen dose tapering during the luteal phase affect the clinical outcome after hormone replacement treatment-frozen-thawed embryo transfer (HRT-FET) cycles? SUMMARY ANSWER: Our results suggest that tapering oestrogen doses during the luteal phase results in similar clinical outcomes to those obtained with the traditional luteal phase support (LPS). WHAT IS KNOWN ALREADY: Traditional LPS with oestrogen and progesterone is considered necessary in HRT-FET cycles. However, case reports have shown successful clinical pregnancies and live births in the absence of oestrogen administration after embryo transfers. STUDY DESIGN, SIZE, DURATION: This was a retrospective study on 6035 HRT-FET cycles extending over 7 years from January 2011 to June 2018 at the reproductive medicine centre of Xiangya Hospital. PARTICIPANTS/MATERIALS, SETTING, METHODS: We compared the clinical outcomes of 1632 HRT-FET cycles with tapered oestrogen doses from 12 days after embryo transfer (study group) to those of 4403 HRT-FET cycles maintained on constant oestrogen doses during the luteal phase (control group) in the case of positive serum HCG test. MAIN RESULTS AND THE ROLE OF CHANCE: We found similar biochemical pregnancy rates (52.1% vs. 51.9, P = 0.864), clinical pregnancy rates (44.9% vs. 43.2%, P = 0.249), implantation rates (29.8% vs. 29.3%, P = 0.591) and miscarriage rates (16.0% vs. 14.6%, P = 0.379) between the studied groups. LIMITATIONS, REASONS FOR CAUTION: Retrospective, design-associated biases are possible. In addition, some baseline characteristics differed between groups. Finally, we did not compare live birth rates between groups. WIDER IMPLICATIONS OF THE FINDINGS: Our study showing similar outcomes between traditional LPS and oestrogen tapering during the luteal phase indicates that oestrogen may be cautiously tapered during the luteal phase after HRT-FET cycles. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the National Natural Science Foundation of China (grant no. 81401269) and the class General Financial Grant from the China Postdoctoral Science Foundation (grant no. 2017M620360). The authors declare that they have no competing interests. TRIAL REGISTRATION NUMBER: N/A.

Hetong decoction relieves loperamide-induced constipation in rats by regulating expression of aquaporins.

OBJECTIVE: To investigate whether Hetong decoction (, HTT) alleviates constipation via regulating AQPs expression. METHODS: Constipation in rats was induced by loperamide, and rats were randomly assigned into model (saline), HHT-low (95 g/kg), HTT-medium (190 g/kg), HTT-high (380 g/kg) and positive control (mosapride) groups. Then the defecation function, the concentration of serum arginine vasopressin (AVP) and cyclic adenosine monophosphate (cAMP), and the expression of AQP3 and AQP8 in colon tissues were assessed. NCM460 colon cells with AQP3 and AQP8 knockdown or overexpression were exposed to serum from rats that received low or high dose of HTT, followed by detection of AQP3 and AQP8 expression. RESULTS: The model group showed lower fecal weight and water content, weaker intestinal transit, higher serum concentration of AVP and cAMP, increased proximal and distal AQP8 expression, increased proximal but decreased distal AQP3 expression. However, these trends were reversed in both the HTT group (low, medium and high dose) and the positive control group. In NCM460 cells, HTT dose-dependently stabilized AQP3 and AQP8 expression under AQP3/8 plasmid interference or overexpression. CONCLUSIONS: HTT relieves constipation in rats through regulating AQP3 and AQP8 expression.

Phylogenetic analysis of the fecal flora of the wild pygmy loris.

The bacterial diversity in fecal samples from the wild pygmy loris was examined with a 16S rDNA clone library and restriction fragment length polymorphism analysis. The clones were classified as Firmicutes (43.1%), Proteobacteria (34.5%), Actinobacteria (5.2%), and Bacteroidetes (17.2%). The 58 different kinds of 16S rDNA sequences were classified into 16 genera and 20 uncultured bacteria. According to phylogenetic analysis, the major genera within the Proteobacteria was Pseudomonas, comprising 13.79% of the analyzed clone sequences. Many of the isolated rDNA sequences did not correspond to known microorganisms, but had high homology to uncultured clones found in human feces.

The association between telomere length and sensitivity to apoptosis of HUVEC.

OBJECTIVES: To study the association between the mean telomere length (MTL) of human umbilical endothelial cells (HUVEC) and their sensitivity to apoptosis. METHODS: Apoptosis of HUVEC was induced by using free hydroxyl radicals. The rate of apoptosis was determined and mean telomere length of HUVEC that were cultured for 1 or 3 months were measured by Southern Blot. RESULTS: At 0.2 mmol/l FeSO(4)/0.0001 mmol/l H(2)O(2) free radical concentration, the apoptosis rate was 8.0 and 17.5% and MTL was 4.66 +/- 0.05 and 3.40 +/- 0.46 kb for HUVEC cultured for 1 and 3 months, respectively. At 0.2 mmol/l FeSO(4)/0.005 mmol/l H(2)O(2), apoptosis rates were 17.4 and 36.0% and MTL were 3.67 +/- 0.06 and 2.90 +/- 0.20 kb for HUVEC cultured for 1 and for 3 months, respectively. Control HUVEC had apoptosis rates of 0.5 and 1.0% and MTL of 5.43 +/- 0.45 and 4.57 +/- 0.21 kb for 1 and 3 months, respectively. The MTL and the apoptosis rates in the treatment groups differed significantly from the controls (p <0.05). CONCLUSIONS: HUVEC with less culture time or short telomere were sensitive to oxidation stress. Oxidation stress also can enhance the shortening of telomere length.

Therapeutic Effects of VEGF Gene-Transfected BMSCs Transplantation on Thin Endometrium in the Rat Model.

OBJECTIVE: Bone mesenchymal stem cells (BMSCs) transplantation has a therapeutic effect on the thin endometrium in animal researches and clinical trials. The present study aims at assessing whether transplantation of VEGF-transfected BMSCs (VEGF-BMSCs) have a better therapeutic effect on endometrial regeneration and endometrial receptivity compared with BMSCs therapy alone. METHODS: Sprague-Dawley (SD) rats were used in the study. Thin endometrium model was established with 95% ethanol injection into uterine. VEGF-BMSCs or BMSCs was transplanted via tail vein IV injection. Endometrial thickness, morphology, and pinopodes were assessed by hematoxylin and eosin (HE) staining and scanning electron microscope (SEM). The proteins and mRNAs expressions of markers for endometrial cells and endometrial receptivity were measured after treatment. The fertility testing was done to assess the embryo implantation efficiency. RESULTS: VEGF-BMSCs transplantation significantly increased endometrial thickness compared with the BMSCs group and the control group. There was no significant difference in endometrial thickness between VEGF-BMSCs group and sham operation group. Importantly, in protein level, expressions of cytokeratin, vitamin, VEGF, LIF, and integrin ανß 3 in VEGF-BMSC group were increased dramatically compared with those of the control group and BMSC group both 4 days and 8 days after stem cells transplantation. Accordingly, mRNA expression of LIF and integrin α ν ß 3 was significantly upregulated compared with those of the control group and BMSC group both 4 and 8 days after treatment. The pinopodes were developed better in the VEGF-BMSCs group and the sham operation group compared with BMSCs group and the control group. The number of embryo implantation is largest in the sham operation group, followed by VEGF-BMSCs group, BMSCs group, and the control group. CONCLUSIONS: Transplantation of VEGF gene-transfected BMSCs may be a better therapeutic treatment for thin endometrium than stem cell therapy alone.

Involvement of the TGFß1/Smad2/MMP3 signaling pathway in SB431542-induced inhibition of cell invasion in multiple myeloma RPMI 8226 cells.

Multiple myeloma (MM) is a malignancy characterized by plasma cell hyperplasia. The majority of patients with MM suffer from mortality due to tumor recurrence and metastasis, which has become an emerging clinical problem. Transforming growth factor ß1 (TGFß1) has been implicated in tumor metastasis; however, its role in RPMI 8226 cells remains to be elucidated. In the present study, RPMI 8226 cells were treated with various concentrations of SB431542, a TGFß1 inhibitor, for 12, 24 and 48 h. RPMI 8226 cells were transfected with lentiviral-TGFß1 vectors to overexpress TGFß1. Cell proliferation rate was subsequently determined by cell-counting kit-8 assay and cell invasion was assessed by Transwell assay. Expression of TGFß1, SMAD family member 2 (Smad2) and matrix metallopeptidase 3 (MMP3) were analyzed by western blotting. The results demonstrated that cell proliferation and invasion of RPMI 8226 cells was significantly inhibited by SB431542 (P<0.05). SB431542 was able to significantly downregulate the expression of TGFß1, phosphorylated (p)-Smad2 and MMP3; however, the overexpression of TGFß1 significantly upregulated the expression of TGFß1, p-Smad2 and MMP3. In conclusion, SB431542 reduced cell invasion in RPMI 8226 cells, and this effect may be mediated via the TGFß1/Smad2/MMP3 signaling pathway.

Establishment and validation of an enzyme-linked immunosorbent assay for IgG antibody against cytomegalovirus based on pp150 antigen.

The development of HCMV vaccines for the prevention of congenital HCMV has been identified as a top priority by the Institute of Medicine (USA), and virus infection is an important endpoint for the efficacy evaluation of the candidate vaccines. However, it is technically difficult to capture the HCMV viremia or uremia in infected individuals because the viremia or uremia can be detected only transiently during acute infection, and most people who are infected are asymptomatic. Thus, it is much desired to develop a serological assay for effectively distinguishing anti-HCMV antibodies as a result of natural infection from those elicited by subunit antigen vaccination. In this study, five HCMV proteins other than antigens commonly used as subunit vaccine candidates were expressed in Escherichia coli and purified, and out of them, the HCMV tegument protein pp150 exhibited the most robust reactivity to seropositive sera and the faintest cross reaction to seronegative sera. With a coefficient of variability less than 15%, an ELISA based on pp150 antigen (pp150-ELISA) showed 100% (366/366) sensitivity and 100% (77/77) specificity (using neutralizing activity as a gold standard) and good quantitative correlation with an in-house ELISA based on virus lysate antigens. Taken together, these results indicate that pp150-ELISA, which has comparable performance to generally used assays based on virus lysate antigens, might provide a simple method to detect HCMV infection or reactivation and could be useful in future HCMV subunit vaccine efficacy trials.

Iron overload increases osteoclastogenesis and aggravates the effects of ovariectomy on bone mass.

Postmenopausal osteoporosis is a metabolic disease associated with estrogen deficiency. The results of numerous studies have revealed the positive correlation between iron accumulation and postmenopausal osteoporotic status. Although the results of previous studies have indicated that estrogen or iron alone have an effect on bone metabolism, their combined effects are not well defined. Using an in vivo mouse model, we found that bone mass was minimally affected by an excess of iron in the presence of estrogen. Once the source of estrogen was removed (ovariectomy), iron accumulation significantly decreased bone mass. These effects were accompanied by fluctuations in the level of oxidative stress. To determine whether these effects were related to bone formation or bone resorption, primary osteoblasts (OBs), RAW264.7 cells, and bone-marrow-derived macrophages were used for in vitro experiments. We found that iron accumulation did inhibit the activity of OBs. However, estrogen had little effect on this inhibition. In contrast, iron promoted osteoclast differentiation through the production of reactive oxygen species. Estrogen, a powerful reactive oxygen scavenger, suppressed this effect in osteoclasts. Our data provided direct evidence that iron affected the bone mass only in the absence of estrogen. The inhibitory effect of estrogen on iron-induced osteopenia was particularly relevant to bone resorption rather than bone formation.

Effects of angiopoietin-1 on inflammatory injury in endothelial progenitor cells and blood vessels.

Endothelial progenitor cells (EPCs) and angiopoietin-1 (Ang-1) play important roles in vasculogenesis and angiogenesis, respectively. Thus, targeting both aspects of cardiovascular tissue regeneration may offer promising therapeutic options for cardiovascular disorders. To this end, we constructed a lentiviral vector (pNL) with the Ang-1 gene and transfected EPCs with it (Ang-1-EPCs) to investigate vasculogenesis in both cellular and animal models. Compared to controls, intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) increased significantly in both untreated EPCs and in the pNL vector group. After Ang-1 transcription, ICAM-1 and VCAM-1 decreased considerably in those treatment groups. Ang-1-modified EPCs alleviated inflammatory responses induced by tumor-necrosis factor-α (TNF-α) in vitro. Moreover, Ang-1-EPC implantation inhibited neointimal hyperplasia after balloon catheter injury in rats, dramatically diminishing the intimal-media (I/M) ratio and decreasing the neointimal area. Proliferating cell nuclear antigen expression in the Ang-1-EPC group was lower than the EPC non-treatment group as well, suggesting that Ang-1-EPC improved cell survival during inflammation and promoted endothelialization in damaged blood vessels.

[In vitro investigation on specific anti-leukemia cell effect of CTL induced by sensitized dendritic cells from umbilical cord blood].

This study was aimed to investigate the specific anti-leukemia cell effect of cytotoxic T lymphocytes (CTLs) induced by HL-60 or K562 cell-sensitized dendritic cells (DCs) from umbilical cord blood. 12 units of human umbilical cord blood (UCB) were collected and the mononuclear cells (MNCs) were isolated from UCB, then cultured with granulocyte monocyte colony- stimulating factor (GM-CSF), interleukin 3 (IL-3), recombinant human stem cell factor (SCF) and EPO for 3 - 4 weeks. Flow cytometry was used to determine the number of DCs and cell surface antigens before and after culture with monoclonal antibodies including CD83, CD1a, CD11c and CDw123. HL-60 and K562 were frozen-thawed, and released their tumor antigen peptides (TAP). The CTLs were produced by sensitizing T lymphocytes with DC-loaded HL-60 and K562 cell antigens. The test of (3)H-TdR incorporation was used to detect the immunostimulation activity of DCs. MTT assay was applied to evaluate specific cytotoxicity of CTL on leukaemia cells. The results indicated that the MNCs of UCBs cultured with GM-CSF, IL-3, EPO and SCF were shown to differentiate into CD1a(+) CD11c(+) CD83(+) CDw123(+) DCs. Numbers of DCs from UCBs remarkably increased in 2 - 4 weeks and then decreased. After culture with cytokines DCs increased (10.6 - 28.2) x 10(5)/ml in actual numbers. The CTL induced by DC pulsed with HL-60, K562 frozen-thawed lysates were effective to kill HL-60 and K562. Cytotoxicity of CTL to HL60 and K562 were (42.04 +/- 8.46)% and (31.25 +/- 11.07)% respectively. It is concluded that the MNCs of UCBs cultured with cytokines of GM-CSF, SCF, EPO and IL-3 can differentiate into CD1a(+), CD83(+), CD11c(+) and CDw123(+) DCs. The CTL induced by DCs pulsed with HL-60, K562 frozen-thawed lysates can effectively kill HL-60 and K562. These DCs as antigen presenting cells play an important role in cancer immunotherapy.

Correlation character of ionic current fluctuations in cell membrane ion channels.

The gating of ion channels has widely been modeled by assuming the transition between open and closed states is a memoryless process. Nevertheless, the statistical analysis of an ionic current signal recorded from voltage dependence K+ single channel is presented. Calculating the sample autocorrelation function of the ionic current based on the digitized signals, rather than the sequence of open and closed states durations time. The result is shown existence memory. For difference voltage, the ion channel current fluctuation has difference correlation attributions. The result suggests the correlation character of the ionic current fluctuations. Also the possible biological implication of the findings have been discussed.