Objective To analyze published literature about the clinical studies on elective single versus double embryo transfer using meta-analysis, so as to provide more convincing evidence for the clinical application of elective single embryo transfer. Methods We electronically searched foreign and domestic biomedical databases including PubMed, Ovid, EMbase, MEDLINE and CENTRAL, to collect randomized controlled trials (RCTs) on elective single versus double embryo transfer. According to Cochrane systematic review method, two reviewers independently screened studies according to inclusion and exclusion criteria, extracted data, and assessed methodological quality of the included studies. Then, meta-analysis was performed using RevMan 5.1 software. Results A total of 9 RCTs involving 2 784 cases were included, of which, 1 452 were in the trial group while the other 1 332 were in the control group. The results of meta-analysis indicated that, compared with elective double embryo transfer, during each transfer period elective single embryo transfer reduced the live birth rate (RR=0.66, 95%CI 0.59 to 0.73, Plt;0.000 01), multiple pregnancy rate (RR=0.05 95%CI 0.02 to 0.11, Plt;0.000 01), preterm birth rate (RR=0.39, 95%CI 0.26 to 0.60, Plt;0.000 1), and low birth weight rate (RR=0.25, 95%CI 0.15 to 0.44, Plt;0.000 01). However, it had no effect on the ectopic pregnancy rate (RR=0.55, 95%CI 0.11 to 2.77, P=0.47), miscarriage rate (RR=1.33, 95%CI 0.92 to 1.91, P=0.13), and neonatal mortality rate (RR=0.31, 95%CI 0.03 to 2.76, P=0.29). Conclusion Compared with elective double embryo transfer, during each transfer period elective single embryo transfer reduces the live birth rate, multiple pregnancy rate, preterm birth rate, and low birth weight rate. No significant difference was found between the two groups in the other indicators.
Objective To systematically review the effectiveness of letrozole combined with GnRH antagonist for in vitro fertilization-embryo transfer (IVF-ET) in poor responders. Methods Such databases as VIP, CNKI, PubMed, EMbase and FMJS were electronically searched for randomized controlled trials (RCTs) or quasi-RCTs on the effectiveness of letrozole combined with GnRH antagonist for IVF-ET. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data, and assessed methodological quality. Then, meta-analysis was performed using RevMan 5.0 software. Results Six studies involving 977 patients were finally included. The results of meta-analysis showed that, for IVF-ET poor responders, compared with the control group, the letrozole combined with GnRH antagonist group had less dosage of Gn (MD=–8.05, 95%CI –13.67 to –2.43, P=0.005), and lower serum E2 value on the day of HCG administration (MD= –1 026.41, 95%CI –1 949.61 to –103.20, P=0.03). However, no significant difference was found in the number of ocytes obtained (MD= –0.61, 95%CI –2.41 to –1.19, P=0.51) and clinical pregnancy rates (OR=1.03, 95%CI 0.53 to 2.02, P=0.92) between the two groups. Conclusion As for the effectiveness of impelling-ovulation treatment for IVF-ET in poor responders, letrozole combined with GnRH antagonist is similar to the control scheme in clinical outcomes, but it reduces the dosage of Gn and treatment costs of IVF-ET, which provides another clinical option for poor responders. Due to the limited quantity and quality of the included studies as well as the difference in methodology, we suggest this above conclusion could be taken as a reference for clinical analysis which needs to be further evaluated in its effects.
Objective To evaluate the effectiveness of GnRH antagonist on vitro fertilization-embryo transfer (IVF-ET). Methods We searched CBMdisc (1979 to 2010), Wanfang (1994 to 2010), CNKI (1994 to 2010), VIP (1989 to 2010), PubMed (1997 to 2010), PML (1997 to 2010), FMJS (2000-2010), and 9 related journals to identify randomized controlled trials (RCTs) on the comparison between GnRH antagonist (GnRHA) and GnRH agonist (GnRHa). The quality of included trials was critically appraised. RevMan 4.2.7 software was used for statistical analysis. Results Six published RCTs involving 1 208 participants were included. Compared with the GnRHa group, stimulation duration in the GnRHA group was lower (WMD= –1.07, 95%CI –1.38 to –0.76), dose of gonadotrophins (Gns) in the GnRHA group was slightly lower (WMD= –0.49, 95%CI –1.63 to 0.66), endometrial thickness at the time of HCG administration was no significant difference in the two groups (WMD= –0.09, 95%CI –0.42 to 0.24), number of oocytes retrieved in the GnRHA group was lower (WMD= –1.80, 95%CI –2.48 to –1.12), OHSS rate in the GnRHA group was slightly lower (Peto OR= 0.77, 95%CI 0.35 to 1.72), pregnancy rate in the GnRHA group was slightly lower (Peto OR= 0.83, 95%CI 0.65 to 1.05), miscarraige rate as no significant difference in the two groups (Peto OR= 1.49, 95%CI 0.79 to 2.82). Conclusions Compared with GnRHa, GnRHA requires shorter stimulation duration and less Gn, less affected the pregnancy rate, and reduces the incidence of OHSS. The use of GnRHA in clinical practice is relatively flexible with good acceptability. GnRHA for the superovulation IVF-ET offers an alternative treatment. The above conclusion still needs more well-designed, multi-center, and large-scale RCTs to confirm and update.
Human SW480 colonic cancer cell line was evaluated for its growth response to pentagastrin, gastrin receptor antagonist proglumide (PGL) in vitro by MTT assay and flow cytometry. The results showed that gastrin possessed a proliferative effect on SW480 cell, PGL alone had no obvious effect on SW480 cell, but it inhibited gastrin-induced growth of SW480 cell with dosage dependent when it was used with gastrin, its inhibitive effect did not steadly increase at a dose＞32μg/ml. This suggests that effect of gastrin is achieved via gastrin receptor. Gastrin promoted the sythesis of DNA, protein and triggered the cancer cell shifting from phase G0/G1 to phase S, G2M. PLG inhibited the effect of gastrin, it suggests that gastrin possessed a proliferation on SW480 cell at post receptor is achieved by the effect of gastrin on cell cycle.
Human SW480 colonic cancer cell line was evaluated for its growth response to Octa peptide somatostatin (SMS 201·995, SMS) in vitro by MTT assay and flow cytometry. The results showed that SMS possessed an inhibitive effect on SW480 cell at dose 1.563－200ng/ml, the maximal effective dose was 50ng/ml. Inhibitive effect of SMS did not steadily increase at a dose ＞50ng/ml. It suggests that effect of SMS is achieved via somatotatin receptor. SMS obviously inhibited the synthesis of DNA and protein, and prohibited the SW480 cell shifting from phase G0/G1 in phase S, G2M, which suggests that somatostatin (SS) possessed an inhibitive effect on large intestinal at cancer cell, it is achieved at receptor by inhibiting the synthesis of DNA and protein and prohibiting cell cycle of cancer.
Objective To observe the morphology and growing status of mesenchymal stem cells(MSCs) of human bone marrow in vitro, in order to confirm that MSCs of human bone marrow are ideal seed cells and provide basic theory for further MSCs research. Methods The methods of density gradient centrifugation with lymphocyte separation medium for human and adherent filtration were used to isolate and purify MSCs of human bone marrow. We observed the cellular growth status and morphology of the primary MSCs and the surface antigens of second passage MSCs were tested. Results The primary culture cells fused into monolayer after 14-16 d. The passage cells kept the same morphological characteristics of primary culture cells. Ultrastructure of the second passage MSCs showed that the shape of nuclei was irregular, there were multiple nucleoli in some of the nuclei, and morphological differentiation of intracytoplasm organelles was immature. The growth curve of the first, fifth and tenth passage cells showed a logarithmic growth at day 3, a peak growth at day 5, and no clones occurred after tenth passage. Cloning efficiency of first passage, fifth passage and tenth passage was respectively 25.83%±2.93%, 14.67%±1.63% and 4.67%±0.52%. Test of MSCs phenotypic characteristics showed a high homogeneity among the cells and surface antigen profiles were positive for CD29, CD44 and negative for CD34, CD45. Conclusion The methods of density gradient centrifugation with lymphocyte separation medium for human and adherent filtration are simple, economic and efficient to isolate and purify MSCs from human bone marrow. With a high proliferating ability in vitro, MSCs from human bone marrow are ideal seed cells for tissue engineers.
Objective To investigate the effects of different temperatures on the system of in vitro physiological environment fostering limbs. Methods Twenty-four limbs were harvested from 6 adult Bama mini pigs and were randomly divided into 4 groups (n=6) according to different temperatures: limbs were placed in in vitro physiological environment foster-ing limbs at 26℃ (group A), 4℃ (group B), 10℃ (group C), and 18℃(group D). After 12 hours of perfusion, the morphology observation was done for the structure and ultrastructure changes of the skeletal muscle by light microscope and transmission electron microscope. The mRNA levels of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) were detected by real-time fluorescent quantitative PCR (RT-qPCR). Results Histological results showed that the skeletal muscle exhibited mild edema, integrity of the sarcolemma, and occasional perivascular inflammatory cell infiltration in groups B, C, and D, meanwhile, the cells of group C had normal morphology; however, muscle fibers degenerated, muscle cells were seriously damaged, a great number of inflammatory cells infiltrated in the fractured muscle fibers in group A. Transmission electron microscope results showed as follows: the muscle fibers arranged in disorder, and many focal solubility necrosis occurred in group A; the muscle fibers arranged in order relatively and sarcolemma was still intact, with mild swelling and flocculent degenerative mitochondria in group B; a large number of muscle fibers arranged in order and regularity with clear sarcomere in group C; and the muscle fibers arranged in disorder and irregularity and partly dissolved in group D. RT-qPCR results showed that the expressions of inflammatory factor TNF-α and IL-1β mRNA in group A were significantly higher than those in groups B, C, and D (P lt; 0.05); the expressions were significantly lower in groups B and C than in group D, and in group C than in group B (P lt; 0.05). Conclusion In the system of in vitro physiological environment fostering limbs, temperature plays an important role in the preservation of amputated limbs. It is suggested that 10℃ can significantly attenuate the reperfusion-induced skeletal muscle cell injuries in this system.
Objective To establish a rapid, simple, and economic method to prepare osteoporosis (OP) in vitro model. Methods Eighty pairs of fresh goat femur were collected from 18-month-old female goats and were randomly divided into 4 groups (20 pairs in each group). The femur was immersed decalcifying solution (18% EDTA) for 1-5 days (group B), 6-10 days (group C), and 11-15 days (group D), while group A had no treatment as control. Four pairs of femur were taken out every day. Quantitative computed tomography was used to scan the medial and lateral femoral condyles, and the bone mineral density (BMD) was calculated. Electronic universal testing machine was used to do three-point bending test and compress and tensile ultimate strenght test, and the mechanical parameters for femur were calculated. Results With demineralized time passing, BMD of the medial and lateral femoral condyles were downtrend in groups A, B, C, and D, showing significant differences among 4 groups (P lt; 0.05); BMD of the lateral femoral condyle was significantly higher than that of the medial femoral condyle in each group (P lt; 0.05). The three-point bending test showed that broken load, ultimate strength, and elastic modulus of groups A and B were significantly higher than those of groups C and D (P lt; 0.05); but no significant difference was found between groups A and B, and between groups C and D (P gt; 0.05). Compress and tensile ultimate strength test showed that the compress and tensile ultimate strengths were significantly higher in group A than in groups C and D (P lt; 0.05), and in group B than in group D (P lt; 0.05), but no significant difference was found between groups A and B, between groups B and C, and between groups C and D (P gt; 0.05). Conclusion The 18% EDTA immersing for 6-15 days is a fast, simple, economical method to prepare an OP in vitro model of goat femur.
Objective In vivo, the microenvironment of epidermal stem cells (ESCs) is complex, and estrogen might be involved in the micro environment. To investigate the biological effects of estrogen on the prol iferation and migration of ESCs in vitro. Methods hESCs were isolated from normal human foreskin and cultured. The second generation of hESCs were identified with flow cytometry after being marked with integrin β1, cytokeratin 19 (CK19), CK14, and CK10 antigens.hESCs of 2 × 106 cell density were cultured with ESCs special medium supplemented with 0.1 nmol/L Diethylstilbestrol in group A (estrogen group), with ESCs special medium supplemented with 10 nmol/L Raloxifene hydrochloride in group B (ER blocking agent group), and with ESCs special medium in group C (control group), respectively. The 100 μm “scratch” wounds were created by scraping confluent hESCs plated on Petri dishes with a sterile pipette tip in vitro. The migrating cells from the wound edge were quantified at 24, 48, and 72 hours after incubation. The rates of wound heal ing were calculated by SigmaScan Pro 5.0 software at 72 hours. The prol iferating effect of estrogen on hESCs was determined with MTT method at 24, 48, 72, 96, and 120 hours. Results Cultured primary hESCs could adhere to the wall showing ovoid in shape and grew into colonies. Flow cytometry showed the positive results for integrin β1, CK19, and CK14 (with positive rate of 96.63%, 95.47%, and 94.27%, respectively) and the negative result for CK10 (with positive rate of 1.32%). In group A, the number of cells crossing the wound edge was more than those of group B and group C at 24, 48, and 72 hours. The rates of wound heal ing were 69.00% ± 0.05% in group A, 35.00% ± 0.05% in group B, and 48.00% ± 0.06% in group C at 72 hours, showing significant differences among groups (P lt; 0.05). The prol iferating speed of hESCs was significantly higher in group A than in groups B and C (P lt; 0.01), and significantly higher in group C than in group B (P lt; 0.01) at 24, 48, 72, 96, and 120 hours. Conclusion The estrogen can promote the prol iferation and migration of hESCs in vitro. It may be involved in many biological activity of skin.
Objective Titania and Ag containing nano-hydroxyapatite/polyamide 66 (TiO2-Ag-nHA/PA66) composite bone fill ing material has good biocompatibil ity and biological safety. To investigate the antibacterial effect and Ag+ release characteristics of TiO2-Ag-nHA/PA66 composite bone fill ing material containing different concentrations of Ag+ in vitro. Methods The n-HA/PA66 composite bone fill ing material A1 (material A1) was prepared by co-polymerization method, and TiO2-Ag-nHA/PA66 composite bone fill ing materials A2 and A3 (materials A2 and A3) were prepared by thesame way containing Ag+ of 0.22wt% and 0.64wt%, respectively, and the TiO2 content was 2.35wt%. The materials A2 and A3 were respectively immersed in 50 mL simulated body fluid (SBF), and Ag+ concentration was measured by atomic absorption spectrometry at 1, 3, 7, 14, 21, and 49 days. The inhibition ring test and colony count method were used to evaluate antibiotic effect against Staphylococcus aureus and Escherichia coli, the anti-adhesion capacity of Staphylococcus aureus and Escherichia coli was observed by scanning electron microscope (SEM). Results There was no significant difference in the Ag+ concentration between materials A2 and A3 at 1 day and 3 days (P gt; 0.05); and there were significant differences in the Ag+ concentration between materials A2 and A3 after 7 days (P lt; 0.05). The inhibition ring diameters of materials A2 and A3 to Staphylococcus aureus and Escherichia coli reached the maximum at 1 day, which were (13.40 ± 2.88), (9.40 ± 1.14) mm and (23.60 ± 1.14), (18.80 ± 0.84) mm, showing significant difference (P lt; 0.05) between materials A2 and A3 respectively; and then, the diameter of inhibition ring reduced with the time. The antibacterial effect of materials A2 and A3 against Staphylococcus aureus and Escherichia coli lasted 15, 33 days and 9, 24 days, respectively. No inhibition ring was observed around material A1 all the time. And the inhibitory rates of materials A2 and A3 were 89.74% ± 3.62%, 94.18% ± 2.05% and 78.65% ± 5.64%, 85.96% ± 2.50%; showing significant differences (P lt; 0.05) among materials A1, A2, and A3. SEM showed that bacterial adhesion of materials A2 and A3 was obviously fewer than that of material A1. Conclusion TiO2-Ag-nHA/PA66 composite bone fill ing material has antibacterial property against Staphylococcus aureus and Escherichia coli, and it has a good release effect in SBF.