The in-vitro fertilization and embryo transfer technique has been widely applied in human insemination. The rate of successful insemination is gradual y rising, and the in-vitro fertilization directly determine the insemination outcome.
OBJECTIVE:To evaluate the difference between the two common using insemination methods, microdrop and open, in in-vitro fertilization and embryo development.
METHODS:A randomized study was conducted to compare microdrop and open insemination methods among non-male factor patients undergoing in-vitro fertilization and embryo transfer. A total of 1 175 cases were enrol ed in the research. There were 573 cases in the microdrop group, and 602 cases in open insemination group. The fertilization rate and embryo development in the two groups were compared.
RESULTS AND CONCLUSION:The fertilization failure rate [total fertilization failure rate+low fertilization rate (<25%oocytes fertilized)] in the microdrop insemination group was higher than in the open insemination group (11.9%, 3.3%, P<0.001), while the good quality embryo rate and pregnancy rate did not differ significantly between the two groups (al P>0.05). The open insemination method is a simple insemination method with a lower fertilization failure rate. As the fertilization is a highly complicated process involving many extrinsic and intrinsic factors, further study is needed to confirm the effects of the two insemination methods on in-vitro fertilization outcome.

Objective To investigate whether recombinant human endostatin can create a time window of vascular normalization prior to vascular pruning to alleviate hypoxia in Lewis lung carcinoma in mice. Methods Kinetic changes in morphology of tumor vasculature in response to recombinant human endostatin were detected under a confocal microscope with immunofluorescent staining in Lewis lung carcinomas in mice. The hypoxic cell fraction of different time was assessed with immunohistochemical staining . Effects on tumor growth were monitored as indicated in the growth curve of tumors . Results Compared with the control group vascularity of the tumors was reduced over time by recombinant human endostatin treatment and significantly regressed for 9 days. During the treatment, pericyte coverage increased at day 3, increased markedly at day 5, and fell again at day 7. The vascular basement membrane was thin and closely associated with endothelial cells after recombinant human endostatin treatment, but appeared thickened, loosely associated with endothelial cells in control tumors. The decrease in hypoxic cell fraction at day 5 after treatment was also found. Tumor growth was not accelerated 5 days after recombinant human endostatin treatment. Conclusions Recombinant human endostatin can normalize tumor vasculature within day 3 to 7, leading to improved tumor oxygenation. The results provide important experimental basis for combining recombinant human endostatin with radiation therapy in human tumors.

Objective To investigate the effect of mitogen-activated protein kinase(MAPK)pathway on the transcriptional expression of mdr1 gene induced by doxorubicin ( DOX)and study the transcription regulation of mdr1 gene.Methods K562 cells were treated with DOX(0.01 μg/ml)with the initial concentration of 0.01 μg/ml for 24 hours,then change the culture media without DOX.K562 cells were cultured until the its status wag recovered.Subsequently the cells were treated with DOX(0.02μg/ml)for 24 hours again.The concentration of DOX was increaged until 0.05 μg/ml by following the protocol above.K562 cells were collected at the concentration of 0.01 μg/ml,0.03μg/ml and 0.05μS/ml DOX.Expression of mdr1 gene were examined by reverse transcription-polymerase chain reaction(RT-PCR).Pglycoprotein(P-gP)wag detected by flow cytometry.Western blot wag performed to detect ERK and P-ERk.K562 cells were pretreated with MAPK inhibitor PD98059 for 1 hour.and then DOX was added.RT-PCR and FCM were used to detect the expression of mdr1 mRNA and P-gp.Results When K562 cells were exposured to DOX.the phosphorylation of ERK wag increaged.the mdr1 gene wag highly expressed as well as its corresponding protein P-gp.When the concentration of DOX was 0.05μg/ml,the expression of mdr1 gene and P-gp were increased over 5 fold.When K562 cells were pretreated with MAPK inhibitor PD98059,the expression of mdr1 gene induced by DOX(the concentration was 0.03 μg/ml and 0.05 μg/m1)was effectively inhibited by(74.1±0.11)%and(70.2±0.14)%respectively.Conclusions DOX could induce the expression of mdr1 gene in K562 cells accompanied by the activation of MAPK/ERK pathway.The block of activation of ERK could inhibit the induced expression of mdr1 gene.

Objective:To explore application value of dance movement therapy in the chemotherapy of young and middle-aged patients with breast cancer, so as to provide reference for rehabilitation nursing.Methods:By convenient sampling method, 90 young and middle-aged female breast cancer patients during chemotherapy from June 2020 to June 2021 in Renmin Hospital of Wuhan University were enrolled in the present study. They were assiged to experimental group and control group with 45 cases in each group according to the enrolled ward. The control group received routine nursing and the experimental group received 4 cycles of dance movement therapy. Before and after intervention, the effects were assessed by Cancer Fatigue Scale (CFS) and Patient-Generated Subjective Global Assessment (PG-SGA) as well as biochemical nutrition indexes.Results:After intervention, the physical fatigue score, emotional fatigue score, cognitive fatigue score and total CFS score were (8.29 ± 3.58), (7.74 ± 1.68), (5.57 ± 1.11), (21.59 ± 4.41) points in the experimental group, which were significantly lower than (9.86 ± 3.49), (8.95 ± 2.62), (6.27 ± 1.70), (25.09 ± 4.33) points in the control group ( t values were 2.07-3.71, all P<0.05). After intervention, the PG-SGA score was (2.81 ± 0.71) points in the experimental group, which was significantly lower than (3.29 ± 1.15) points in the control group ( t=2.37, P<0.05). Conclusions:Dance movement therapy can alleviate the cancer related fatigue and promote nutritional status of young and middle-aged female breast cancer patients with chemotherapy.

Objective: To clone mouse rod opsin promoter (ROP) and establish transgenic mice harboring mouse rod opsin promoter and enhanced green fluorescent protein(mROP-EGFP) fusion gene. Methods: Mouse ROP was cloned from C57BL/6 mouse genomic DNA by polymerase chain reaction (PCR). Expression vector of mROP-EGFP fusion gene were constructed by recombination DNA technique. It was identified by restriction endonucleases digestion and confirmed by DNA sequencing. After Not I restriction endonuclease digestion, the coding elements were microinjected into male pronuclei of mice zygotes to generate transgenic mice. The pups were evaluated by PCR at genomic DNA level and mated with normal mouse. Expression of GFP in retina of transgenic mice was detected by fluorescent microscope. Results: 2. 1 kb mouse rod opsin promoter fragment was amplified from mice genome DNA. Expression vector pmROP-EGFP was constructed successfully. Following microinjection of coding sequence of pmROP-EGFP, 3 pups were verified to integrate the mROP-EGFP fusion gene in their genomic DNA by PCR assay, named C57-TgN (mROP-EGFP )SMMU21, C57-TgN (mROP-EGFP)SM-MU26 and C57-TgN(mROP-EGFP) SMMU27. They could express GFP in retina. Conclusion: 2. 1 kb mouse rod opsin promoter is cloned and expression vector pmROP-EGFP is constructed. mROP-EGFP fusion gene transgenic mice are established, which harboring mROP-EGFP gene and expressing GFP in their retina. This is valuable for studying the development of brain and retina, pathogenesis of retina disorder and retina transplanting.

Aim To investigate how chili-derived exosome-like nanovesicle(CDELN)inhibited ox-LDL uptake and reduced ox-LDL-induced intracellular cholesterol accumulation.Methods CDELN were isolated and purified using tissue crushing,differential centrifugation,ultracentrifugation and sucrose density gradient centrifugation.Ox-LDL was used to stimulate THP-1-derived macrophages for 24 hours to establish a foam cell model in vitro,and the effect of CDELN on macrophage foam and its mechanism were further studied.Confocal laser microscopy was used to detect the uptake of CDELN and DiL-acetylated low density lipoprotein(DiL-ac-LDL)by THP-1 macrophages.Oil red O staining was used to detect intracellular cholesterol content and the positive area of oil red 0 staining in cells was analyzed to evalu-ate the effect of intracellular lipid accumulation.RT-qPCR and Western blot were used to detect mRNA and protein levels of scavenger receptor A(SRA)and cluster of differentiation 36(CD36),lectin-like oxidized low density lipoprotein recep-tor-1(LOX-1),ATP-binding cassette transporter A1/G1(ABCA1/G1).The expression of mitogen-activated protein ki-nase(MAPK)pathway proteins including p-ERK,p-p38 MAPK and p-c-Jun,were also analyzed.Results CDELN were exosome-like nanovesicles with uniform size and double membrane,rich in protein and nucleic acids,which can be taken up by macrophages.The results of DiL-ac-LDL uptake showed that CDELN could inhibit cholesterol uptake of mac-rophages.Oil red 0 staining showed that CDELN could reduce ox-LDL-induced intracellular cholesterol accumulation.RT-qPCR and Western blot showed that CDELN could significantly reduce mRNA levels of matrix metalloprotein-9(MMP-9),SRA,CD36 and LOX-1 and protein levels of p-ERK,SRA,CD36 and LOX-1 in ox-LDL-induced THP-1-derived macro-phages.Treatment with the p-ERK agonist Yoda1 diminished the protective effect of CDELN.Conclusion CDELN can significantly inhibit macrophage foam cell formation,and this effect may be associated with the inhibition of phosphoryl-ation levels of ERK1/2 in the MAPK pathway.

AIM:To predict the mechanism of Sanguis draconis flavones(SDF)in the prevention and treat-ment of myocardial ischemia reperfusion injury(MIRI)based on network pharmacology and molecular docking methods.METHODS:The main chemical constituents of SDF were collected through literature search,and the targets of key con-stituents were screened by using the SwissTargetPrediction and TargetNet databases.Disease targets were also screened based on GeneCards,OMIM,TTD and PharmGkb databases,then targets were intersected with Cytoscape to construct the"drug-key constituent-target"network diagram,and the core target was obtained through visualization and analysis by Cytoscape software.Gene Ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were analyzed by the Metascape platform.By utilizing AutoDock Vina software and Pymol,molecular docking between core compounds and core targets was carried out.Further,animal experiments were performed to explore the pharmacodynamic mechanism of SDF.RESULTS:The active constituents of SDF included loureirin B and loureirin A,which were mapped to 391 targets.A total of 3 096 MIRI disease targets were obtained from the database,af-ter intersection,172 intersection targets were obtained,and 56 core targets were acquired through analysis.The core relat-ed pathways included the cancer pathway and cell death signaling pathway.The results of molecular docking verified the strong binding activity between key constituents and key targets.Animal experiments demonstrated that SDF effectively prevented and treated MIRI,significantly inhibited the arachidonic acid 15-lipoxygenase(ALOX15)mRNA and protein expression,and reduced the myocardial infarction size after MIRI.CONCLUSION:SDF may play a positive role in the treatment of MIRI,which may be related to the regulation of the ALOX15 factor.

Purpose: The risk stratification of pediatric anaplastic large cell lymphoma (ALCL) has not been standardized. In this study, new risk factors were included to establish a new risk stratification system for ALCL, and its feasibility in clinical practice was explored. Materials and Methods: On the basis of the non-Hodgkin’s lymphoma Berlin–Frankfurt–Munster 95 (NHL-BFM-95) protocol, patients with minimal disseminated disease (MDD), high-risk tumor site (multiple bone, skin, liver, and lung involvement), and small cell/lymphohistiocytic (SC/LH) pathological subtype were enrolled in risk stratification. Patients were treated with a modified NHL-BFM-95 protocol combined with an anaplastic lymphoma kinase inhibitor or vinblastine (VBL). Results: A total of 136 patients were enrolled in this study. The median age was 8.8 years. The 3-year event-free survival (EFS) and overall survival of the entire cohort were 77.7% (95% confidence interval [CI], 69.0% to 83.9%) and 92.3% (95% CI, 86.1% to 95.8%), respectively. The 3-year EFS rates of low-risk group (R1), intermediate-risk group (R2), and high-risk group (R3) patients were 100%, 89.5% (95% CI, 76.5% to 95.5%), and 67.9% (95% CI, 55.4% to 77.6%), respectively. The prognosis of patients with MDD (+), stage IV cancer, SC/LH lymphoma, and high-risk sites was poor, and the 3-year EFS rates were 45.3% (95% CI, 68.6% to 19.0%), 65.7% (95% CI, 47.6% to 78.9%), 55.7% (95% CI, 26.2% to 77.5%), and 70.7% (95% CI, 48.6% to 84.6%), respectively. At the end of follow-up, one of the five patients who received maintenance therapy with VBL relapsed, and seven patients receiving anaplastic lymphoma kinase inhibitor maintenance therapy did not experience relapse. Conclusion This study has confirmed the poor prognostic of MDD (+), high-risk site and SC/LH, but patients with SC/LH lymphoma and MDD (+) at diagnosis still need to receive better treatment (ClinicalTrials.gov number, NCT03971305).

Tooth enamel is prone to be attacked by injurious factors, leading to a de/remineralization imbalance. To repair demineralized enamel and prevent pulp inflammation caused by biofilm accumulation, measures are needed to promote remineralization and inhibit bacterial adhesion on the tooth surface. An innovative material, poly (aspartic acid)-polyethylene glycol (PASP-PEG), was designed and synthesized to construct a mineralizing and anti-adhesive surface that could be applied to repair demineralized enamel. A cytotoxicity assay revealed the low cytotoxicity of synthesized PASP-PEG. Adsorption results demonstrated that PASP-PEG possesses a high binding affinity to the hydroxyapatite (HA)/tooth surface. In vitro experiments and scanning electron microscopy (SEM) demonstrated a strong capacity of PASP-PEG to induce in situ remineralization and direct the oriented growth of apatite nanocrystals. Energy dispersive X-ray spectroscopy (EDS), X-ray diffraction analysis (XRD) and Vickers hardness tests demonstrated that minerals induced by PASP-PEG were consistent with healthy enamel in Ca/P ratio, crystal form and surface micro-hardness. Contact angle tests and bacterial adhesion experiments demonstrated that PASP-PEG yielded a strong anti-adhesive effect. In summary, PASP-PEG could achieve dual effects for enamel repair and anti-adhesion of bacteria, thereby widening its application in enamel repair.

Objective    To summarize the experience of surgical treatment of asphyxiating thoracic dysplasia (Jeune syndrome). Methods    A total of 15 patients with asphyxiating thoracic dysplasia from August 2018 to April 2020 in our hospital were retrospectively included. There were 7 males and 8 females, aged 1-25 (8.87±6.71) years. Special steel bars were used to correct the growth direction of the rib and costal cartilage. Meanwhile, the concave and convex deformities of the chest wall on both sides were corrected to increase the chest volume and correct the thoracic deformity. Results    The contour appearance of the chest wall of all patients changed after the operation. The shape was close to normal, and the symptoms of hypoxia were improved. The operation time was 147.73±59.78 min, intraoperative bleeding volume was 105.67±91.90 mL, ICU stay time was 14.20±13.54 d and hospital stay time was 26.00±17.87 d. Eleven patients were directly extubated after the operation, 4 patients underwent tracheotomy and received assisted respiration, and the assisted respiration time was 19, 13, 22 and 12 days, respectively. The postoperative chest circumference was significantly increased, and the blood oxygen saturation was significantly improved. There were 5 patients with cardiac insufficiency, and 3 of them were improved by cardiotonic therapy, 2 of them died of heart failure on the 2nd and 31st day after the operation, respectively. Abdominal distention occurred in 10 patients after operation, and 5 of them were obstinate and eliminated by comprehensive treatment. All patients were followed up. The appearance of thorax was improved obviously and there was no sign of compression in lungs. One 13-year-old patient developed respiratory discomfort 3 months after the operation, and the symptoms were relieved after self-administration of oxygen. A 25-year-old patient developed cardiac insufficiency half a month after the discharge, and the symptoms disappeared after cardiotonic treatment. Four patients took out the steel bars in 13, 13, 15 and 17 months after the operation, respectively. The appearance of thorax remained well after the operation. The imaging examination showed that the position of bone structure was normal, the lung field was clear, and there was no sign of chronic inflammation. Conclusion    This technique is a safe and simple operation method. It can not only eliminate the deformity of chest wall, but also increase the volume of chest obviously. However, the long-term effect needs to be further evaluated.