Plant-derived nanovesicles: Further exploration of biomedical function and application potential

Extracellular vesicles (EVs) are phospholipid bilayer vesicles actively secreted by cells, that contain a variety of functional nucleic acids, proteins, and lipids, and are important mediums of intercellular communication. Based on their natural properties, EVs can not only retain the pharmacological effects of their source cells but also serve as natural delivery carriers. Among them, plant-derived nanovesicles (PNVs) are characterized as natural disease therapeutics with many advantages such as simplicity, safety, eco-friendliness, low cost, and low toxicity due to their abundant resources, large yield, and low risk of immunogenicity in vivo. This review systematically introduces the biogenesis, isolation methods, physical characterization, and components of PNVs, and describes their administration and cellular uptake as therapeutic agents. We highlight the therapeutic potential of PNVs as therapeutic agents and drug delivery carriers, including anti-inflammatory, anticancer, wound healing, regeneration, and antiaging properties as well as their potential use in the treatment of liver disease and COVID-19. Finally, the toxicity and immunogenicity, the current clinical application, and the possible challenges in the future development of PNVs were analyzed. We expect the functions of PNVs to be further explored to promote clinical translation, thereby facilitating the development of a new framework for the treatment of human diseases.

Perioperative management and operative treatment of malignant tumor of anal canal merging severe abdominal protuberance / 中华胃肠外科杂志

Objective: To report the perioperative management and robot-assisted minimally invasive surgery results of one case with malignant tumor of anal canal combined with severe abdominal distention. Methods: A 66-year-old male suffer from adenocarcinoma of anal canal (T3N0M0) with megacolon, megabladder and scoliosis. The extreme distention of the colon and bladder result in severe abdominal distention. The left diaphragm moved up markedly and the heart was moved to the right side of the thoracic cavity. Moreover, there was also anal stenosis with incomplete intestinal obstruction. Preoperative preparation: fluid diet, intravenous nutrition and repeated enema to void feces and gas in the large intestine 1 week before operation. Foley catheter was placed three days before surgery and irrigated with saline. After relief of abdominal distention, robotic-assisted abdominoperineal resection+ subtotal colectomy+colostomy was performed. Results: Water intake within 6 hours post-operatively; ambulance on Day 1; anal passage of gas on Day 2; semi-fluid diet on Day 3; safely discharged on Day 6. Conclusion: Robotic-assisted minimally invasive surgery is safe and feasible for patients with malignant tumor of anal canal combined with severe abdominal distention after appropriate and effective preoperative preparation to relieve abdominal distention.

Roles of TSA upregulation miR-4298 targeting inhibition of PADI4 expression in inducing U251 cells apoptosis / 国际肿瘤学杂志

Objective:To investigate the mechanism of microRNA-4298 (miR-4298) targeting inhibition of peptidylarginine deiminase 4 (PADI4) expression in U251 cells apoptosis induced by histone deacetylase inhibitor trichostatin A (TSA).Methods:The experiment was divided into TSA group (0.2 μmol/L TSA) and contral group. CCK8 method was used to detect the effect of TSA on the proliferation of U251 cells. Flow cytometry was used to detect the apoptosis level of U251 cells after drug action. Reverse transcription PCR and Western blotting experiments were used to determine the changes of PADI4 gene and protein expression after miR-4298 interference. Luciferase assay was used to determine the effects of miR-4298 on targeted binding and luciferase activity in the 3′UTR region of PADI4. U251 cells were transfected with PADI4 to observe the rescue effect of miR-4298 on apoptosis. Each experiment was divided into 3 groups, NC group, miR-4298 mimic group, TSA+ miR-4298 mimic group and NC group, miR-4298 inhibitor group, TSA+ miR-4298 inhibitor group.Results:U251 cells were treated with 0.2 μmol/L TSA for 4 days, the Absorbancy ( A) values of the control group was 1.168±0.148, which was higher than those of the TSA group (0.737±0.007), with statistically significant difference ( t=4.948, P=0.008). Compared with the control group, after treatment with 0.2 μmol/L TSA for 48 h, U251 cells showed obvious apoptosis (27.62%±3.49% vs. 4.99%±0.13%, t=11.190, P<0.001). Compared with those before the drug treatment, the PADI4 gene expression (0.386±0.020 vs. 0.903±0.021) and protein expression (0.276±0.041 vs. 0.777±0.031) after TSA treatment for 48 h both were decreased, with statistically significant differences ( t=30.400, P<0.001; t=16.770, P<0.001). The expression of miR-4298 during the process of TSA-induced U251 cell apoptosis increased (2.573±0.289 vs. 1.003±0.136; t=8.487, P=0.001). There was a negative correlation between the miR-4298 expression and PADI4 expression ( r=-0.877, P=0.002). Luciferase experiment confirmed that miR-4298 has targeted binding and luciferase inhibitory effects on the 3′UTR region of wild-type PADI4. Compared with NC group (0.920±0.026), the relative expression levels of PADI4 gene of miR-4298 mimic group (0.413±0.049) and TSA+ miR-4298 mimic group (0.213±0.035) were decreased, with statistically significant differences (all P<0.001). The PADI4 protein expression level change was consistent with the gene change trend. Compared with NC group (3.78%±0.68%), the apoptosis levels of miR-4298 mimic group (7.96%±1.10%) and TSA+ miR-4298 mimic group (13.74%±1.26%) were increased, with statistically significant differences ( P=0.005; P<0.001). Compared with NC group (0.183±0.025), the PADI4 gene expression of miR-4298 inhibitor group (0.483±0.032) and TSA+ miR-4298 inhibitor group (0.386±0.025) were increased, with statistically significant differences ( P<0.001; P=0.015). The PADI4 protein expression level change was consistent with the gene change trend. Compared with NC group (4.96%±0.59%), the apoptosis levels of miR-4298 inhibitor group (23.83%±2.20%) and TSA+ miR-4298 inhibitor group (9.55%±1.49%) were increased, with statistically significant differences (all P<0.001). In the rescue experiment, the expression level of PADI4 in miR-4298 mimic group was significantly increased ( P<0.001), while the expression level of PADI4 in miR-4298 mimic+ PADI4 group was relatively reversed ( P=0.002). Conclusion:miR-4298 can participate in the process of U251 cell apoptosis by targeting the expression of PADI4 gene, and miR-4298 may be the target of targeted intervention therapy for glioma.

Effect of Metabolic Reprogramming on Toxin Eliminating Therapeutics in Targeting Lung Cancer Stem Cells / 中国实验方剂学杂志

Objective::To observe the effect of realgar nanoparticles (a representative drug in toxin eliminating therapeutics) targeting hypoxia-inducible factors (HIF), which act as effector molecules on metabolic reprogramming of lung cancer stem cells, and to explore the effect mechanism of lung cancer stem cells and metabolic reprogramming in the process of lung cancer metastasis, so as to verify the effectiveness of toxin eliminating therapeutics in the prevention and treatment of lung cancer metastasis. Method::Lung cancer A549 cells were cultured in vitro, and lung cancer stem cells were then identified and selected. The stem cells were divided into blank control group, cisplatin group (5 mg·L-1), realgar nanoparticles low, medium and high dose groups (100, 200, 400 mg·L-1). After intervention, glucose oxidase method was used to detect the effect of realgar nanoparticles on the glucose metabolism of lung cancer stem cells, real-time polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of hypoxia-inducible factors-1α (HIF-1α), C-myc and p53, while Western blot was used to detect the expression of related proteins HIF-1α, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and mammalian target of rapamycin (mTOR), and enzyme linked immunosorbent assay (ELISA) was used to detect the glucose transporter 1 (GLUT1), pyruvate dehydrogenase kinase 1 (PDK1), pyruvate kinase M (PKM), phosphofructokinase(PFK), pyruvate dehydrogenase (PDH) and lactic dehydrogenase (LDH) expression. Result::As compared with the blank control group, realgar nanoparticles can reduce the glucose consumption of lung cancer stem cells, and the glucose consumption was reduced with the increase of dose in a time-and dose-dependent manner (P<0.01). Realgar nanoparticles can inhibit the mRNA expression of HIF-1α, a key factor in metabolic reprogramming of lung cancer stem cells (P<0.05, P<0.01), down-regulated C-myc mRNA and up-regulated the p53 mRNA expression (P<0.05, P<0.01), down-regulated protein expressions of PI3K, Akt, mTOR(P<0.05, P<0.01), and inhibited the expression of related enzymes GLUT1, PDK1, PFK, PKM, PDH, and LDH levels (P<0.05, P<0.01). With the increase of dose, the regulation and control ability of realgar nanoparticles gradually increased. Conclusion::Toxin eliminating therapeutics can drive the metabolic reprogramming of lung cancer stem cells by targeting HIF effector molecule, and then inhibit the invasion and metastasis of lung cancer.

Effect of miR-5581-5p/TRIM22 on acute promyelocytic leukemia cell differentiation / 国际肿瘤学杂志

Objective:To investigate the function of miR-5581-5p and its interaction with tripartite motif 22 (TRIM22) during the terminal differentiation of human acute promyelocytic leukemia (APL) cells into granulocytes.Methods:APL cells (NB4) were differentiated into granulocytes by all-trans retinoic acid (ATRA), using dimethylsulfoxide (DMSO) as the control. The expression of TRIM22 was detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blotting, and the expression of miR-5581-5p was detected by qRT-PCR during cell differentiation. miRNA expression was regulated by cell transfection with miR-5581-5p mimic and inhibitor, and negative control was set, and qRT-PCR was used to verify the regulatory effect. Luciferase binding assay was performed to detect the presence of targeted binding. Western blotting was used to detect the expression of TRIM22 after miRNA differential expression. Flow cytometry was used to detect the effects of the regulation of miR-5581-5p on the differentiation of NB4 cells induced by ATRA.Results:After ATRA induced NB4 cells to differentiate into granulocytes, the gene expression level of TRIM22 was significantly higher than that of the control group (24.56±2.80 vs. 1.02±0.13; t=8.392, P=0.001). The level of protein expression was also significantly higher than that of the control group (0.80±0.01 vs. 0.17±0.01; t=44.900, P<0.001). The expression level of miR-5581-5p in NB4 cells differentiation group was significantly lower than that in the control group (0.14±0.02 vs. 1.01±0.08; t=10.840, P<0.001). The results of the dual luciferase reporter gene showed that the luciferase activity of the co-transfected miR-5581-5p mimic and TRIM22 WT group was significantly lower than that of the co-transfected miR-5581-5p mimic and TRIM22 MUT group (0.73±0.02 vs. 0.98±0.03; t=7.534, P=0.002). Western blotting showed that after transfection with miR-5581-5p inhibitor, the expression of TRIM22 was significantly higher than that of the negative control (0.44±0.01 vs. 0.21±0.01; t=18.290, P<0.001). While after transfection with miR-5581-5p mimic, the expression of TRIM22 decreased significantly compared with the negative control (0.62±0.01 vs. 0.80±0.02; t=6.402, P=0.003). CD11b expression of miR-5581-5p mimic group after ATRA treatment was significantly lower than that of the control group (45.80±1.80 vs. 56.61±1.88; t=4.159, P=0.014). The expression of CD11b in miR-5581-5p inhibitor group was significantly higher than that in the control group (66.48±2.54 vs. 52.60±1.70; t=4.539, P=0.011). Conclusion:miRNA-5581-5p can bind to TRIM22 3′UTR and negatively regulate TRIM22 expression. The decrease of miR-5581-5p can increase the expression of TRIM22, then promote the differentiation of ATRA-induced NB4 cells into granulocytes.

Research Progress and Forensic Identification of Alcoholic Cardiomyopathy / 法医学杂志

With the rapid development of the social economy in China, the incidence of diseases caused by excessive drinking is gradually increasing as well. Alcoholic cardiomyopathy refers to long-term high intake of ethanol, and has typical dilated cardiomyopathy characteristics, such as, hemodynamic changes, symptoms, signs, and morphological features. It is a kind of cardiomyopathy that excludes other causes of dilated cardiomyopathy. Due to the lack of specific pathological changes, the forensic pathological identification of alcoholic cardiomyopathy can only be based on the patient's medical history and by ruling out other causes of cardiomyopathy. This paper reviews the pathogenesis and forensic identification of alcoholic cardiomyopathy in order to provide reference for forensic pathologists and clinicians.

Molecular Mechanism of Belamcandae Rhizoma and Ephedrae Herba Couplet Medicines in Treating Asthma Based on Network Pharmacology / 中国实验方剂学杂志

Objective:To analyze the effective active ingredients of Belamcandae Rhizoma and Ephedrae Herba couplet medicines(BREH)in the treatment of bronchial asthma based on network pharmacology, in order to predict their potential targets and explore the mechanism. Method:Active ingredients and predict their targets were collected from traditional Chinese medicine system pharmacology(TCMSP) database. Drugs-components-targets network and Proteins interations network were built by STRING database and Cytoscape software. ClusterProfiler and ClueGO was used to enrich the biological function and metabolic pathway of core targets. Finally, candidate targets were mapped onto the pictures of correlative pathways. Result:The 38 effectively active ingredients were screened out, including luteolin, stigmasterol, diosmetin, naringenin, quercetin, iristectorigenin A, isorhamnetin. There were 214 candidate targets relating to bronchial asthma, and 55 core ones were selected to be mainly studied, including RAC-alpha serine/threonine-protein kinase (Akt1), tumor necrosis factor (TNF), mitogen-activated protein kinase 1 (MAPK1), vascular endothelial growth factor A (VEGFA), interleukin-10 (IL-10), NF-kappa-B inhibitor alpha (NFKBIA), and a number of relevant gene ontology(GO) functions and Kyoto Encyclopedin of Genes and Genomes(KEGG) pathways were enriched. Conclusion:BREH may regulate the Th1, Th2 and Th17 cell differentiations, Asthma, IL-17, phosphatidylinositol-3-kinases(PI3K)/Akt, MAPK, NF-κB, VEGF signaling pathways, so as to interfere the process of cell metabolism, and inhibit gene expression of proinflammatory factor in the treatment of bronchial asthma.

Platelet-rich plasma combined with conventional surgery in the treatment of atrophic nonunion of femoral shaft fractures: study protocol for a prospective, randomized,controlled clinical trial / 中国组织工程研究

BACKGROUND:Internal and external fixation combined with autologous bone graft for treating atrophic nonunion has a long treatment cycle,and moreover,it cannot achieve a 100％ cure rate.Platelet-rich plasma contains a variety of growth factors and a large number of white blood cells,and contributes to tissue healing.However,there is no clinical study on the effectiveness of platelet-rich plasma combined with conventional surgery in the treatment of atrophic nonunion.OBJECTIVE:To investigate the effectiveness of platelet-rich plasma in the treatment of atrophic nonunion of femoral shaft fractures.METHODS:We conducted a prospective,open-label,randomized,controlled clinical trial at the Affiliated Hospital of Qinghai University,China.Ninety-two patients with atrophic nonunion of femoral shaft fractures were equally and randomly divided into control group and experimental group.Patients in the control group received conventional surgery.Patients in the experimental group were injected with autologous platelet-rich plasma on the basis of conventional surgery.The primary outcome was fracture healing rate at postoperative 9 months.The secondary outcomes were visual analogue scale scores in resting state and during passive motion,healing time,treatment costs,and adverse reactions.The study protocol was approved by the Ethics Committee of Affiliated Hospital of Qinghai University of China (approval number:QHG0223A) on May 20,2014.Written informed consent was provided by each patient and their family members after they fully understood the treatment plan.RESULTS AND CONCLUSION:Our partial results demonstrated that visual analogue scale scores and complications were similar between the two groups at postoperative 1-3 days.The healing rate was significantly higher in the experimental group than in the control group.The healing time was significantly shorter in the experimental group than in the control group.This trial will provide objective data for the clinical use of platelet-rich plasma combined with conventional surgery for the treatment of atrophic nonunion.

Evaluation of the Sensitivity and Specificity of the New Clinical Diagnostic and Classification Criteria for Kashin-Beck Disease, an Endemic Osteoarthritis, in China / 生物医学与环境科学（英文）

This study aimed to evaluate the sensitivity and specificity of the new clinical diagnostic and classification criteria for Kashin-Beck disease (KBD) using six clinical markers: flexion of the distal part of fingers, deformed fingers, enlarged finger joints, shortened fingers, squat down, and dwarfism. One-third of the total population in Linyou County was sampled by stratified random sampling. The survey included baseline characteristics and clinical diagnoses, and the sensitivity and specificity of the new criteria was evaluated. We identified 3,459 KBD patients, of which 69 had early stage KBD, 1,952 had stage I, 1,132 had stage II, and 306 had stage III. A screening test classified enlarged finger joints as stage I KBD, with a sensitivity and specificity of 0.978 and 0.045, respectively. Shortened fingers were classified as stage II KBD, with a sensitivity and specificity of 0.969 and 0.844, respectively, and dwarfism was classified as stage III KBD with a sensitivity and specificity of 0.951 and 0.992, respectively. Serial screening test revealed that the new clinical classification of KBD classified stages I, II, and III KBD with sensitivities of 0.949, 0.945, and 0.925 and specificities of 0.967, 0.970, and 0.993, respectively. The screening tests revealed that enlarged finger joints, shortened fingers, and dwarfism were appropriate markers for the clinical diagnosis and classification of KBD with high sensitivity and specificity.

Effect of betaloc combined with lotensin on cardiac function and NT-proBNP level in patients with chronic congestive heart failure / 药物评价研究

Objective To analyze the effect ofbetaloc combined with lotensin on cardiac function and N-terminal B type natriuretic peptide (NT-proBNP) level in patients with chronic congestive heart failure (CHF).Methods 210 CHF patients in People's Hospital in Qinghai Province from December 2015 to December 2016 were divided into three groups by random number table,70 cases in each.Three group were given routine examination,cardiotonic and diuretic treatment,and on this basis,control A group purely added lotensin,control B group purely added betaloc,observation goup took betaloc combined with lotensin.The cardiac function,exercise tolerance,NT-proBNP level,concentration of hemoglobin (Hb),heart rate,blood pressure,clinical efficacy and safety before and after treatment were compared among the three groups.Results Before treatment,there was no statistical difference in the cardiac function,NT-pro BNP level and Hb content among three groups;After treatment,the cardiac function and exercise tolerance of observation group were significantly better than those of control A and B group (P ＜ 0.05) The NT-pro BNP level,heart rate and blood pressure of observation group were significantly lower than those of control A and B group (P ＜ 0.05) The Hb content of observation group was higher than that of control A and B group (P ＜ 0.05).The total effective rate of observation group was significantly higher than that of control A and B group (P ＜ 0.05).There was no statistical difference in the incidence of adverse reactions among three groups.Conclusion Betaloc combined with lotensin in treatment of CHF can effectively improve cardiac function,relieve heart failure and increase exercise tolerance of patients,which has clinical application value.

Frozen embryo transfer combined with intrauterine administration of autologous peripheral blood mononuclear cells for repeated implantation failure: report of 3 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the efficacy of frozen-thawed embryo transfer combined with intrauterine administration of autologous peripheral blood mononuclear cells (PBMCs) in the treatment of repeated implantation failure (RIF).</p><p><b>METHODS</b>PBMCs obtained from 3 patients with RIF on the day of follicle rupture (natural cycle) or when the endometrial thickness reached 8 mm (hormone replacement cycle) were cultured in the presence of HCG for 48 h. The cultured PBMCs, along with freshly isolated PBMCs, were administered into the uterine cavity of the patients. Vitrified cleavage-stage embryos or blastocysts transfer was performed on day 3 or 5, respectively.</p><p><b>RESULTS</b>Vitrified embryo or blastocyst transfer resulted in pregnancy and healthy live births in all the 3 patients.</p><p><b>CONCLUSION</b>Frozen-thawed embryo transfer combined with intrauterine administration of autologous PBMCs may be an effective and safe approach to the treatment of RIF and may improve the outcomes of assisted reproduction.</p>

Relation between insulin resistance and insulin receptor gene methylation in the endometrium of patients with polycystic ovary syndrome / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between insulin resistance and methylation of insulin receptor (INSR) gene in the endometrium of women with polycystic ovary syndrome (PCOS).</p><p><b>METHODS</b>Based on the HOMA index, 35 patients with PCOS were divided into insulin resistant group (IR group, n=18) and non-resistant group (NIR group, n=18). The patients age, serum estriol, testosterone, FSH and LH, fasting insulin and fasting blood glucose were compared between the two groups. The endometrial samples were obtained from the patients to examine DNA methylation status of INSR gene in the endometrial cells using methylation-specific PCR.</p><p><b>RESULTS</b>The BMI, WHR, fasting glucose, fasting insulin, and HOMA index differed significantly between the two groups (P<0.05). PCR analysis showed partial methylation in the promoter region of INSR gene in 13 samples in IR group and 11 samples in NIR group, without detection of full methylation of the INSR gene in either group. The methylation status showed no significant difference between the two groups (P=0.328).</p><p><b>CONCLUSION</b>Partial methylation of the INSR gene occurs in the endometria of PCOS patients, but this study does not provide a strong evidence supporting the relationship between insulin resistance and INSR gene methylation in women with PCOS.</p>

CA 125 expression in cervical and vaginal secretions in women in normal reproductive period / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the tumor-associated antigen CA125 expression in the serum and cervical and vaginal secretions in women during normal reproductive period, and explore the clinical value of detecting tumor markers in the cervical and vaginal secretions.</p><p><b>METHODS</b>A total of 145 women in reproductive period were divided into 3 age groups (20-29 years, 30-39 years, and over 40 years), and their CA125 levels in cervical secretion, vaginal secretion and serum were detected by automatic electro-chemiluminescent immunoassay.</p><p><b>RESULTS</b>CA125 levels in the cervical secretion, vaginal secretion and serum showed no significant difference between the 3 age groups (P>0.05). In each group, CA125 levels differed significantly between the cervical secretion, vaginal secretion and serum (P<0.001). In the 145 women, the average CA125 level was 497.82 - or + 75.29 U/ml in the cervical secretion, 114.66 - or + 26.40 U/ml in vaginal secretion and 18.06 - or + 3.35 U/ml in serum, showing significant differences between them (P<0.001).</p><p><b>CONCLUSION</b>CA125 expression level is significantly higher in the cervical and vaginal secretions than in the serum in women in normal reproductive period, and its levels in cervical and vaginal secretions can be more sensitive and convenient for early detection of related diseases.</p>

Inhibitory effect of RNA interference targeting BaxBak on apoptosis of human granulosa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of small interfering RNA (siRNA) targeting Bax-Bak on the apoptosis of human granulosa cells.</p><p><b>METHODS</b>Human granulosa cells were transfected with Bax-siRNA and Bak-siRNA either alone or in comibnation, and the cell morphological changes were obsered and the cell apoptosis was detected with flow cytometry. Western blotting was performed to examine the changes in Bax and Bak expressions in the transfected cells.</p><p><b>RESULTS</b>Western blotting demonstrated significantly weakened expressions of Bax and Bak in the transfected cells. The cell morphology of the cells tranfected with Bak siRNA and with both Bak and Bax siRNA remained normal; the cells with exclusive Bax siRNA transfection presented with basically normal cell morphology, but black spots were noted in the cytoplasm. In the positive and negative control groups, the cells became rounded and shrank with expanded intercellular spaces and numerous black spots in the cytoplasm. Flow cytometry showed apoptotic indexes of 3.44% and 3.97% in cells transfected with Bak siRNA and Bax-Bak siRNA, respectively, significantly lower than that in the negative group. Bax siRNA transfection resulted in an apoptotic index of 19.98%, similar to that in the negative group.</p><p><b>CONCLUSION</b>Interference of the expression of Bak gene inhibits the apoptosis of human granulosa cells, and the inhibitory effect can be enhanced by simultaneous Bax interference, which, when used alone, does not obviosuly inhibit the apoptosis of human granulosa cells.</p>

Clinical application of prefilled pen and conventional syringe during controlled ovarian stimulation for in vitro fertilization / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the efficacy, convenience and costs of recombinant follitropin alpha administered by a prefilled pen device and conventional syringe in Chinese women undergoing controlled ovarian stimulation for in vitro fertilization (IVF).</p><p><b>METHODS</b>A total of 184 patients undergoing IVF treatment were enrolled in this study. According to a long-term recombinant follicle-stimulating hormone (rFSH) protocol, ovarian stimulation was performed with the prefilled pen and conventional syringe at random in these subjects, and the dose of follitropin, number of oocytes and embryo parameters and IVF-ET outcome were compared between the two groups.</p><p><b>RESULTS</b>The total rFSH dose, cost, and frequency of hospital visits were significantly lower in the pen protocol group, but the residual rFSH amount was higher. Compared with conventional injections, the prefilled pen was associated with significantly lowered rate of local redness, high rate of local bruise, more frequent follitropin dose modulation and lower serum oestradiol levels on HCG day. No significant difference was found in the endometrial thickness, numbers of oocytes retrieved, MII oocytes, transferred embryo, or the clinical pregnancy rates between the two groups. The ratio of MII oocytes, good quality embryo rates and implantation rates was significantly higher in the pen group with lower incidences of moderate and severe ovarian hyperstimulation syndrome.</p><p><b>CONCLUSION</b>The prefilled pen provides an easy, safe, effective and more patient-friendly means for controlled ovarian stimulation procedure in Chinese women, but more attention should be given to protocol optimization and patient education.</p>

Relationship between protein oxidation levels in the follicular fluid and the outcome parameters of in vitro fertilization-embryo transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the relationship between protein oxidation levels in the follicular fluid and the outcome parameters of in vitro fertilization-embryo transplantation (IVF-ET).</p><p><b>METHODS</b>The levels of advanced oxidation protein products (AOPP) in the follicular fluid were measured in 64 women with tubal infertility undergoing IVF-ET. The relationship between the AOPP levels and IVF-ET outcome parameters was analyzed.</p><p><b>RESULTS</b>AOPP levels showed significant inverse correlations between the proportion of mature oocytes (r=-0.401, P=0.001), fertilization rate (r=-0.257, P=0.045), cleavage rate (r=-0.290, P=0.024) and good embryo rate (r=-0.520, P=0.000). AOPP levels differed significantly between the groups with different retrieved oocyte numbers (F=3.851, P=0.027), being the lowest in women with 8 to 15 retrieved oocytes and the highest in those with retrieved oocytes below 8. The AOPP level in the non-pregnant women was significantly higher than that in the pregnant women (t=3.665, P=0.001). The AOPP levels also differed significantly with age (F=15.919, P=0.000), and the women >35 years of age had the highest level and those below 30 years had the lowest level.</p><p><b>CONCLUSION</b>Protein oxidative stress is present in the follicular fluid of women on IVF-ET cycles. High level of AOPP may have adverse effects on the oocytes and early embryonic development and may affect the outcome of IVF-ET.</p>

Expression of trophinin in human oocytes and preimplantation embryos / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the significance of trophinin expression in human oocytes and preimplantation embryos.</p><p><b>METHODS</b>The expression of trophinin in 9 human oocytes, 16 blastomeres and 12 blastocysts were detected by indirect immunofluorescence assay and observed under laser scanning confocal microscope.</p><p><b>RESULTS</b>Human oocytes, blastomeres and blastocysts were all positive for trophinin expression, and the positivity intensified significantly in the course of the embryonic development (P<0.05).</p><p><b>CONCLUSION</b>Trophinin may play an important role in human embryo implantation by mediating homophilic adhesion between the embryo and the endometrium during the implantation window.</p>

Proteomics of follicular fluid in mature human follicles and antral follicles: a comparative study with laser desorption/ionization-time of flight-mass spectrometry / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the differentially expressed proteins between mature and antral follicles and identify the proteins crucial for follicle development and oocyte fertilization.</p><p><b>METHODS</b>Mature follicular fluids were obtained from 48 women after oocyte collection during in vitro fertilization (IVF), and antral follicular fluids were collected from 21 women by follicular puncture. The proteins in the follicular fluids were detected by surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) and weak cation-exchange protein chip (WCX-2). The data were read with PBSII-C type protein chip reader and analyzed with Biomarker Wizard and Biomarker Patterns Software of Ciphergen Company.</p><p><b>RESULTS</b>In comparison with those in the antral follicular fluid, the concentration of 4 proteins were significantly different in mature follicular fluid, including two up-regulated and two down-regulated proteins.</p><p><b>CONCLUSION</b>Significant variation occurs in the proteomics of mature follicular fluid, and the differentially expressed proteins may have close relation to follicle development and oocyte fertilization.</p>

F10 gene knock-down mediated by RNA interference induces apoptosis of KLE cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of short double-stranded RNA (dsRNA) on F10 gene expression in KLE cells and the effect of F10 knock-down on KLE cell apoptosis.</p><p><b>METHODS</b>The short dsRNA specifically targeting F10 gene prepared by in vitro transcription was transfected into KLE cells via lipofectamine 2000. The expression of F10 mRNA in the transfected KLE cells was detected by real-time quantitative RT-PCR, and the apoptosis of the cells was assayed by flow cytometry.</p><p><b>RESULTS</b>Real-time quantitative RT-PCR demonstrated that transfection of the KLE cells with the short dsRNA induced effective knock-down of F10 gene, and transfection of the cells with 20 nmol/L dsRNA for 48 h decreased the expression of F10 mRNA by 83%. Compared with the control, the apoptosis index of the transfected KLE cells increased from 0.36% to 8.91%.</p><p><b>CONCLUSION</b>F10 gene in KLE cells can be specifically knocked-down with dsRNA prepared by in vitro transcription, and the down-regulation of F10 gene induces apoptosis of KLE cells.</p>

Allogeneic leukocyte immunization combined with IVF-ET for treatment of infertility induced by recurrent spontaneous abortion / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of allogeneic leukocyte immunization combined with in vitro fertilization-embryo transfer (IVF-ET) for treatment of infertility induced by habitual abortion.</p><p><b>METHODS</b>Allogeneic leukocyte immunization was performed in 9 patients with infertility induced by habitual abortion, with another 9 patients undergoing IVF-ET without habitual abortion as the control group. All the patients were treated with long GnRH-a protocols. The infertility patients with recurrent spontaneous abortion history were immunized with lymphocytes from the husband for before IVF-ET and after clinical pregnancy.</p><p><b>RESULTS</b>The fertilization rates of the immunotherapy group and control group were 81.3% and 82.2%, respectively, showing no significant difference (P>0.05). Five patients in each group had clinical pregnancy, and a twin pregnancy occurred in the control group. The embryo implantation rates were also comparable between the two groups (22.7% vs 28.6%, P>0.05). All the fetuses resulted from IVF-ET developed normally and were healthily delivered.</p><p><b>CONCLUSION</b>Allogeneic leukocyte immunotherapy along with IVF-ET is effective for treatment of infertility resulting from recurrent spontaneous abortion.</p>