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.

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.

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

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

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 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.

The purpose of this study was to investigate the vasorelaxing effect of vasonatrin peptide (VNP) on human intramammary artery (HIMA).The vasorelaxing effect of VNP on HIMA was measured by means of perfusion in vitro. The effects of HS-142-1, TEA, 8-Br-cGMP and methylene blue (MB) were also observed. It was found that VNP caused a concentration-dependent relaxation in HIMA which was independent of the endothelium. 8-Br-cGMP (0.1-1000 micromol/L) also caused a concentration-dependent relaxation in HIMA. The vasorelaxing effect of VNP disappeared in the presence of HS-142-1 (20 micromol/L), an antagonist of the natriuretic peptide guanylate cyclase (GC) receptor. MB (10 micromol/L), an inhibitor of GC, not only blocked completely the relaxation of HIMA, but also enhanced the vascular contraction induced by norepinephrine. TEA (1 mmol/L), an antagonist of calcium activated potassium channels (K(Ca)), reduced but not completely blocked the vasorelaxing effect of VNP. These findings suggest that VNP can relax HIMA, which is independent of the endothelium. This effect is possibly achieved by the binding of VNP with the natriuretic peptide GC receptors in the smooth muscle cells (SMCs), leading to an increase in intracellular cGMP level. Moreover, the vasorelaxing effect of VNP is associated with K(Ca).
Aged ; Atrial Natriuretic Factor ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; In Vitro Techniques ; Mammary Arteries ; drug effects ; physiology ; Middle Aged ; Potassium Channels, Calcium-Activated ; metabolism ; Receptors, Guanylate Cyclase-Coupled ; metabolism ; Vasodilation ; drug effects ; physiology

OBJECTIVETo study the prognostic significance of grading system for stromal invasion in pathologic tumor stage T1 (pT1) adenocarcinoma of lung.

METHODSEighty-five cases of surgically resected pT1 lung adenocarcinoma with clinicopathologic and follow-up data were retrospectively reviewed. The degree of invasive growth was classified into three grades according to its location in the tumor. The clinicopathologic characteristics and prognostic significance were analyzed.

RESULTSAmongst the 85 cases studied, 17 cases (20%) were in grade 1, 12 (14%) in grade 2 and 56 (66%) in grade 3. The tumor size was smaller and lymphovascular permeation was less frequently encountered in cases with grade 1 stromal invasion than in those with grade 3 (P=0.005 for tumor size and P=0.018 for occurrence of lymphovascular permeation). The rate of lymph node metastasis and pathologic staging in cases with grade 1 and grade 2 were similar and were significantly lower than those with grade 3 (P=0.007 for rate of lymph node metastasis in grade 1 versus grade 3 tumors, P=0.002 for pathologic stage in grade 1 versus grade 3 tumors, P=0.027 for rate of lymph node metastasis in grade 2 versus grade 3 tumors and P=0.021 for pathologic stage in grade 2 versus grade 3 tumors). There was no statistically significant difference with respect to age, gender and smoking history of the patients, amongst cases in different grades. The overall five-year survival rate was 63%. The five-year survival rates for cases with grade 1, grade 2 and grade 3 were 100%, 83.3% and 46.6%, respectively. The difference between cases with grade 2 and grade 3 was statistically significant (P=0.027). The death rate during follow-up for cases with grade 1, grade 2 and grade 3 were 0, 16.7% and 42.9%, respectively. The difference between cases with grade 1 and grade 3 was statistically significant (P=0.001). Univariate analysis showed that grade of stromal invasion (P=0.001), pathologic stage (P<0.001), presence of lymphovascular permeation (P<0.001) and lymph node involvement (P<0.001) represented important prognostic factors. Multivariate analysis also showed that pathologic stage (P<0.001) was an independent prognostic factor.

CONCLUSIONSThe grading system of stromal invasion in pulmonary adenocarcinoma correlates with tumor prognosis and other prognostic factors. It represents a useful criterion in prognostic categorization of pT1 adenocarcinoma of lung.

Adenocarcinoma ; pathology ; Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; pathology ; Neoplasm Staging ; methods ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate

OBJECTIVETo observe the expression of Toll-like receptor 8 (TLR8) in human cervical cancer cell-line HeLa cells, and the effects of TLR8 agonist CL075 on the survival and proliferation of HeLa cells.

METHODSPCR and RT-PCR were used to detect the expression of TLR8 in 13 cancer cell lines, and the expression of COX-2, Bcl-2, VEGF mRNA in the HeLa cells stimulated by TLR8 agonist CL075 were also measured by RT-PCR. Immunofluorescence technique was used to determine the exact location of TLR8 in the cells. The percentage of viable cells was determined by trypan blue exclusion after the HeLa cells were stimulated with TLR8 agonist CL075 (0.1 µg/ml, 0.5 µg/ml, 1.0 µg/ml, 2.5 µg/ml), and cell cycle and apoptosis were analyzed by flow cytometry, and the proliferation was measured by MTT.

RESULTSCompared with the other cancer cell lines, the expression of TLR8 in HeLa cells was the highest (703.7 ± 20.6). After stimulation by CL075, the cells had a remarkable increase of the percentage of cells in G(2)/M + S phases. In the control group, the percentage of cells in G(2)/M +S phases was (39.02 ± 2.33)%, whereas after stimulated with 1.0 µg/ml CL075, the percentage of cells in G(2)/M + S phases reached the highest ratio (57.67 ± 1.73)%, and the percentage of cells in G(2)/M + S phases had a less decrease after 2.5 µg/ml CL075 stimulation and the percentage was (56.14 ± 3.73)%. After the CL075 treatment, there was no significant changes of apoptosis compared with that of the control cells (P > 0.05), but after DDP treatment the apoptosis had a significant change (P < 0.01). After stimulation by 1.0 µg/ml CL075 for 24 h, no significant difference (P > 0.05) was found by MTT test, but a significant difference was found at 48 h and 72 h (P < 0.01). An increased expression of COX-2, Bcl-2 and VEGF mRNA was observed in HeLa cells after stimulation by TLR8 agonist CL075 for 24 h and 48 h (P < 0.05).

CONCLUSIONSExpression of TLR8 is significantly increased in HeLa cells. The proportion of cells at different phases has a significant change after CL075 stimulation, which may up-regulate the proliferation of HeLa cells. These data suggested that TLR8 agonist may influence the tumor development and TLR8 may become a potential target in the treatment for cervical cancer.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Cyclooxygenase 2 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; HeLa Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Quinolines ; pharmacology ; RNA, Messenger ; metabolism ; Thiazoles ; pharmacology ; Toll-Like Receptor 8 ; agonists ; genetics ; metabolism ; Up-Regulation ; Vascular Endothelial Growth Factor A ; genetics ; metabolism