Intracytoplasmic sperm injection,which combined with in vitro fertilization and micromanipulation techniques,has been applied to research the molecular mechanisms of fertilization,and produce the sexing livestock and transgenic animals.The research advances in porcine intracytoplasmic sperm injection,including in vitro maturation and pretreatment of oocytes,selection and treatment of spermatozoon,artificial activation of injected oocytes after injection,and improvement of the operation technique were reviewed.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Fluorescence ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; isolation & purification ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Tuberculosis, Pulmonary ; diagnosis ; microbiology ; Young Adult

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L⁻¹ permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L⁻¹ permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L⁻¹ permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L⁻¹ permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L⁻¹ permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L⁻¹ compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L⁻¹ permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L⁻¹ permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L⁻¹ permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L⁻¹ permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

Objective To evaluate the diagnostic accuracy of 3.0 T contrast enhanced (CE) whole heart coronary MRA ( CE MRA ) using 32-channel coils with high acceleration factor. Methods Sixty patients with suspected coronary artery disease who were scheduled for coronary angiography (CAG)underwent CE CMRA at 3.0 T MRI scanner. A 32-channel receiver coil was used for data acquisition. For image acquisition, an ECG-triggered, navigator-gated, inversion-recovery prepared, segmented gradient-echo sequence was used with an acceleration factor of three in the phase-encoding direction using GRAPPA reconstruction. Gd-BOPTA (0.15 mmol/kg body weight) was intravenously administered at a rate of 0. 3 ml/s. The diagnostic accuracy in detecting significant stenoses ( ≥50% of vessel lumen) was evaluated using χ2 test with X-ray angiography as the reference. Results Whole-heart CE CMRA was successfully completed in 56 patients who were scheduled for CAG. The averaged imaging time was ( 6. 0 ± 1.3 ) min.3.0 T CE CMRA using 32 channel coils correctly identified significant CAD in 28 patients and correctly ruled out CAD in 23 patients. The sensitivity and specificity were 93. 3% and 88.5% respectively.Conclusion Combined with dedicated 32-channel coils, 3.0 T CE CMRA allows significant reduction in imaging speed and reduced dose of the contrast agent. These improvements resulted in substantially improved overall accuracy of CE CMRA in detecting coronary artery disease.

Objective To explore the detection methods for cognitive dysfunction of the major depression in Elderly and analyze their clinical significance.Methods Using matched-pairs study,42 patients with seniie de- pressive disorders(experimental group)and 42 normal aged people(control group)were examined with auditory e- voked potential P300(event related potential,ERP-P300)and SECF,respectively.Results It was found that the scores with registration,span,recall,classification and total score of the subjects in the experimental group were sig- nificantly lower than those in the control group(P

Objective To evaluate the value of contrast-enhanced whole-heart coronary magnetic resonance angiography ( CE CMRA ) at 3.0 T in the delineation of cardiac venous anatomy. Methods Contrast-enhanced whole-heart CMRA at 3.0T was performed in 43 consecutive subjects using ECG-triggered, navigator-gated, inversion-recovery prepared, segmented gradient-echo sequence with a 32-channel cardiac coil. The visibility of the coronary veins was graded visually using a 4-point scale.Continuous variable was expressed as (-x)±s. The paired student t test was used to evaluate the differences of the coronary sinus (CS) ostium diameter in anteroposterior and superoinferior directions. Results CMRA examination was successfully completed in 40 subjects with acquisition time of ( 6. 9 ± 1.8 ) min. The cardiac veins were finally evaluated in 38 of 40 (95.0%) subjects. The mean distance of the posterior vein of the left ventricle (PVLV) and the left marginal vein (LMV) to the CS ostium were (3.34 ± 0. 90) and (6. 12 ± 1.02) cm, respectively. The mean visibility scores of CS, posterior interventricular vein (PIV),PVLV, LMV, and anterior interventricular vein (AIV) were 4.0 ± 0.0, 3.4 ± 0. 5, 3.4 ± 0. 5, 3.0 ± 0. 8,and 3. 3 ± 0. 5, respectively. The diameter of the CS ostium in the superoinferior direction ( 1.13 ±0. 26) cm was larger than that in the anteroposterior direction (0. 82 ± 0. 19) cm (t = -4. 31 ,P <0. 05).Conclusion Contrast-enhanced whole-heart CMRA at 3.0 T can clearly depict the cardiac venous anatomy.

Objective To investigate the association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms with metabolic syndrome among Kazakh and Han ethnicities in Xinjiang.Methods PCR-RFLP was used to detect 802 subjects' lipoprotein lipase Hind Ⅲ and S447Xgenotypes (including 201 controls and 200 metabolic syndrome patients in Kazakh and Han ethnicities, respectively). Results (1) Frequencies of H + H-/H-H- genotype (32.50% vs.47.76%), H- allele( 18.00% vs. 28.86%), SX/XX genotype (8.00% vs. 22.39%) and X allele (4.00%vs. 12.44% ) for metabolic syndrome in Hah ethnicity were all significantly lower than those in controls (P＜ 0.01 ). (2) The frequencies of H + H-/H-H- genotype (33.50% vs. 46.80% ), H- allele (22.00% vs. 28.60%), SX/XX genotype (10.50% vs. 22.90%) and X allele (5.50% vs. 12.44% ) in patients with metabolic syndrome in Kazakh were all significantly lower than those for controls (P＜0.01). (3) The frequencies of lipoprotein lipase gene Hind Ⅲ and S447X genotypes and alleles in Kazakh were not significantly different from Han (all P＞0.05). (4)The levels of waist circumference, systolic blood pressure, diastolic blood pressure, triglyceride and FPG in H + H-/H-H- and SX/XX genotype were significantly lower than those in H + H + and SS genotype.HDL-C was significantly higher than that in H + H + and SS genotype (P＜0.05). (5) The frequencies of H + H + and SS genotype increased along with the increase in number of metabolic syndrome component. Conclusion The lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms were associated with metabolic syndrome risk in Kazakh, and H + H-/H-H- genotype, H- allele, SX/XX genotype and X allele might have served as protective factors of metabolic syndrome. H + H-/H-H- and SX/XX genotype seemed to have had beneficial effects for all the metabolic syndrome components, and the frequencies of H + H + and SS genotype were increasing along with the increase of number in the metabolic syndrome components.

Adult ; Female ; Humans ; Male ; Sturge-Weber Syndrome ; diagnosis ; pathology ; Young Adult

OBJECTIVETo explore whether there are gene mutations of Tolloid-like 1 (TLL-1) gene in Chinese patients with sporadic congenital heart disease (CHD).

METHODSOne hundred and fifteen patients with sporadic CHD were selected as CHD group. One hundred and two age and gender-matched healthy people were recruited as control group. After amplifying the exon 10 of the TLL-1 gene by polymerase chain reaction, the polymerase chain reaction products were purified, sequenced and analyzed in order to investigate the TLL-1 gene mutation.

RESULTSAn insertion mutation of base A in the exon 10 of TLL-1 gene was identified in 7 out of 115 CHD patients, including 3 patients with atrial septal defect, 2 patients with ventricular septal defect, 1 patients with patent ductus arteriosus and 1 patients with complex CHD, the total mutation rate was 6.1% in CHD group and 0 in control group (P < 0.01).

CONCLUSIONSTLL-1 gene mutation with an insertion mutation of base A in exon 10 is often in Chinese patients with various CHD. The underlying pathogenesis between TLL-1 gene mutation and occurrence of congenital heart disease in Chinese people remains unclear and warrants further investigations.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; Exons ; Female ; Heart Defects, Congenital ; genetics ; Humans ; Male ; Middle Aged ; Mutagenesis, Insertional ; Pedigree ; Tolloid-Like Metalloproteinases ; genetics ; Young Adult

The Spindle-view, a specialized instrument for observing spindle image, was applied to observe the meiotic spindles of vitro matured porcine oocytes at 36, 42, 44, 48h, and enucleation from porcine, comparing to the previously methods (McGrath-Solter's method and two-step-squeezing method) in the enucleated. The results showed that: (1) there was no noticeable differences at vicinity of spindle images and 1st polar body among in vitro matured porcine oocytes at 40-48 h under the instrument; (2) Spindle-view is suitable for the observation of meiotic spindles of matured oocytes and enucleation from porcine; the modified Spindle-view method for enucleation is significantly better than McGrath-Solter' s method and two-step-squeezing method in the enucleated rates (95.5%, 42.1%, 74.2%, P < 0.0l) of absolutely removing nuclei matter; (3) the spindle images could be used to monitor the oocyte qualities.
Animals ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; Cytological Techniques ; Female ; Nuclear Transfer Techniques ; Oocytes ; cytology ; Spindle Apparatus ; ultrastructure ; Swine