OBJECTIVETo explore the application value of morphology assessment of sperm from fresh semen in routine in vitro fertilization (IVF).

METHODSWe analyzed the morphology of the sperm from fresh or optimized semen samples and, based on the sperm morphology of the raw semen, allocated 908 IVF cycles due to the pure tubal factor to different groups: morphologically normal sperm (MNS) ≤ 4%, > 4% - ≤ 15%, and > 15% in Trial 1 and MNS ≤ 1%, > 1% - ≤ 2%, > 2% - ≤ 3%, and > 3%-- ≤ 4% in Trial 2. We compared the rates of fertilization, cleavage, high-quality embryo, -blastocyst formation, and pregnancy among different groups.

RESULTSThe total fertilization rate was significantly lower in the MNS ≤ 4% than in the MNS > 4% - ≤ 15% and >15% groups (74.40% vs 78.61% and 80.03%, P < 0.01). Compared with the MNS ≤ 1%, > 1% - ≤ 2%, and > 2% - ≤ 3% groups, the MNS > 3% - ≤ 4% group showed remarkably increased rates of 2PN normal fertilization (77.23%, 78.97% and 78.99% vs 85.47%, P < 0.01), cleavage (95.71%, 96.01% and 97.27% vs 98.73%, P < 0.05), and blastocyst formation (53.85%, 49.01% and 49.55% vs 63.41%, P < 0.01). No statistically significant differences were observed in the rates of clinical pregnancy, implantation, early abortion, live birth, or malformation at birth among different groups (P > 0.05).

CONCLUSIONMNS ≤ 4% affected the total rate of fertilization while MNS ≤ 3% reduced the rate of normal fertilization in IVF. However, even MNS ≤ 1% did not result in fertilization disorder or failure. Therefore, teratozoospermia alone was not an indicator of ICSI and sperm mor- phology assessment had no obvious value for predicting the rates of embryo quality, clinical pregnancy, and live birth in IVF.

Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Spermatozoa ; cytology

Radiofrequency (RF) radiation at the frequency of mobile phones has been not reported to induce cellular responses in in vitro and in vivo models. We exposed HEI-OC1, conditionally-immortalized mouse auditory cells, to RF radiation to characterize cellular responses to 1763 MHz RF radiation. While we could not detect any differences upon RF exposure, whole-genome expression profiling might provide the most sensitive method to find the molecular responses to RF radiation. HEI-OC1 cells were exposed to 1763 MHz RF radiation at an average specific absorption rate (SAR) of 20 W/kg for 24 hr and harvested after 5 hr of recovery (R5), alongside sham-exposed samples (S5). From the whole-genome profiles of mouse neurons, we selected 9 differentially-expressed genes between the S5 and R5 groups using information gain-based recursive feature elimination procedure. Based on support vector machine (SVM), we designed a prediction model using the 9 genes to discriminate the two groups. Our prediction model could predict the target class without any error. From these results, we developed a prediction model using biomarkers to determine the RF radiation exposure in mouse auditory cells with perfect accuracy, which may need validation in in vivo RF-exposure models.
Absorption ; Animals ; Cellular Phone ; Gene Expression* ; Mice ; Neurons ; Support Vector Machine ; Biomarkers

Heat shock protein 70 (HSP70), which evidences important functions as a molecular chaperone and anti-apoptotic molecule, is substantially induced in cells exposed to a variety of stresses, including hypertonic stress, heavy metals, heat shock, and oxidative stress, and prevents cellular damage under these conditions. However, the molecular mechanism underlying the induction of HSP70 in response to hypertonicity has been characterized to a far lesser extent. In this study, we have investigated the cellular signaling pathway of HSP70 induction under hypertonic conditions. Initially, we applied a variety of kinase inhibitors to NIH3T3 cells that had been exposed to hypertonicity. The induction of HSP70 was suppressed specifically by treatment with protein kinase C (PKC) inhibitors (Go6976 and GF109203X). As hypertonicity dramatically increased the phosphorylation of PKC micron, we then evaluated the role of PKC micron in hypertonicity-induced HSP70 expression and cell viability. The depletion of PKC micron with siRNA or the inhibition of PKC micron activity with inhibitors resulted in a reduction in HSP70 induction and cell viability. Tonicity-responsive enhancer binding protein (TonEBP), a transcription factor for hypertonicity-induced HSP70 expression, was translocated rapidly into the nucleus and was modified gradually in the nucleus under hypertonic conditions. When we administered treatment with PKC inhibitors, the mobility shift of TonEBP was affected in the nucleus. However, PKC micron evidenced no subcellular co-localization with TonEBP during hypertonic exposure. From our results, we have concluded that PKC micron performs a critical function in hypertonicity-induced HSP70 induction, and finally cellular protection, via the indirect regulation of TonEBP modification.
Animals ; Carbazoles/pharmacology ; Cell Line ; Flavonoids/pharmacology ; HSP70 Heat-Shock Proteins/*biosynthesis ; Humans ; Indoles/pharmacology ; Isoquinolines/pharmacology ; MAP Kinase Signaling System/physiology ; Maleimides/pharmacology ; Mice ; NFATC Transcription Factors/metabolism ; Phosphorylation ; Promoter Regions, Genetic ; Protein Kinase C/antagonists & inhibitors/*physiology ; Protein Transport ; Saline Solution, Hypertonic/*pharmacology ; Signal Transduction ; Sulfonamides/pharmacology

Objective To investigate the distribution and clonality of T cell receptor(TCR)Vγand Vδ subfamily in peripheral blood of patients with allergic rhinitis before and after 1 year treatment with immunotherapy.Methods The specific IgE and the complementary determinant region 3(CDR3)of TCR V γ(Ⅰ-Ⅲ)and Vδ(1-8)subfamily genes in mononuclear cells were amplified from 10 effective cases of allergic rhinitis before and after 1 year treatment with immunotherapy,to observe the distribution and utilization of TCR Vγ and Vδ repertoire.The positive PCR products were further labeled with RT-PCR and analyzed by gene scan technique to determine the CDR3 size and evaluate the clonality of the detectable TCR Vγ and Vδ T cells.Peripheral blood of 10 healthy adults served as controls.Results All symptoms were significantly improved after 1 year specific iammanotherapy,but no changes were seen in specific IgE [(22.89 ±9.60)kU/L before treatment,(19.62 ± 7.63)kU/L after treatment,Z =1.051,P ＞ 0.05].No statistically significant differences of expression levels of the TCR Vγ　 Ⅰ-Ⅲ subfamily genes were found between patients with allergic rhinitis normal control group(t value were-0.679,-0.516,-0.808,all P ＞ 0.05),but significantly decreased after 1 year treatment.There were statistically significant differences of expression levels of the TCR VγⅠ-Ⅱ subfamily genes before and after treatment(t value were-2.904,-2.217,all P ＜ O.05).5.30 ± 0.82,4.90 ± 0.57 and 5.20 ± 1.40 out of TCR Vδ(1-8)subfamilies were selectively expressed in T cells in patients with allergic rhinitis before and after 1 year treatment and normal control group,predominantly for TCR Vδ 1,2,3 and 6.The TCR Vδ 6 subfamily was found to have statistically significant differences in these groups(Fisher′s Exact Test,P ＜ 0.05).Compared with the normal control group and the allergic rhinitis group before treatment,a significant higher frequency of Vδ 6 oligoclonal was identified in T cells in patients with allergic rhinitis after 1 year treatment.Conclusions There was difference in the expression levels of the TCR Vγ Ⅰ-Ⅲ subfamily genes and distribution and clonality of TCR Vγand Vδ subfamily T cells in peripheral blood of patients with allergic rhinitis before and after 1 year treatment.Specific immunotherapy can be effective in alleviation of the symptom in patients with allergic rhinitis during the early stage,possibly by inducing TCR γδ T cells,especially the TCR Vδ6 subfamily,and possibly no significant relativity between symptom and specific IgE.

We have investigated biological responses to radiofrequency (RF) radiation in in vitro and in vivo models. By measuring the levels of heat shock proteins as well as the activation of mitogen activated protein kinases (MAPKs), we could not detect any differences upon RF exposure. In this study, we used more sensitive method to find the molecular responses to RF radiation. Jurkat, human T-Iymphocyte cells were exposed to 1763 MHz RF radiation at an average specific absorption rate (SAR) of 10 W/kg for one hour and harvested immediately (R0) or after five hours (R5). From the profiles of 30,000 genes, we selected 68 differentially expressed genes among sham (S), R0 and R5 groups using a random-variance F-test. Especially 45 annotated genes were related to metabolism, apoptosis or transcription regulation. Based on support vector machine (SVM) algorithm, we designed prediction model using 68 genes to discriminate three groups. Our prediction model could predict the target class of 19 among 20 examples exactly (95% accuracy). From these data, we could select the 68 biomarkers to predict the RF radiation exposure with high accuracy, which might need to be validated in in vivo models.
Absorption ; Apoptosis ; Cellular Phone ; Heat-Shock Proteins ; Humans ; Jurkat Cells* ; Metabolism ; Mitogen-Activated Protein Kinases ; Support Vector Machine* ; Biomarkers

OBJECTIVETo investigate the clinical type, diagnosis and prognosis of endoscopic surgery of choanal polyp (CP).

METHODSThirty four cases of CP treated between January 1998 to December 2005 were retrospectively reviewed. Pathogenesis, original sites, clinical manifestation, its relationship with paranasal sinuses, methods and effects of endoscopic surgery on CP were analysed.

RESULTS(1) In this study, 18 cases originated from the cyst or polyp in sinus (among them, 17 from maxillary sinus, 1 from posterior ethmoid sinus). (2) Five cases from nasal fontanelles or sphenoethmoidal recess, mucosa of sphenoidal ostium, accompanied with homolateral empyema or mucosal edema of maxillary sinus or sphenoid sinus. (3) Eleven cases from middle turbinate, uncinate process, olfactory cleft, nasal septum and anterior wall mucosa of ethmoidal sinus with normal adjacent sinus. (4) All cases were treated by complete excision of the CP and open sinus procedures of the affected ones with intranasal endoscopic approach. No recurrence was observed during the follow-up.

CONCLUSIONS(1) We propose to divide the CP into three clinical types: intrasinus, sinus obstruction and simple ones. We also suggest to abide by the operation principal based on clinical types and to select rational methods and extent of operation. (2) Definite diagnosis and clinical type can be obtained by endoscopic examination and CT scan of sinus before operation. (3) Endoscopic surgery is precise and mini-invasive in treatment of CP and complete removal of CP root can effectively reduces the recurrence.

Adolescent ; Adult ; Aged ; Child ; Endoscopy ; Female ; Humans ; Male ; Maxillary Sinus ; Middle Aged ; Nasal Polyps ; diagnosis ; surgery ; Otorhinolaryngologic Surgical Procedures ; methods ; Retrospective Studies ; Young Adult

Adult ; Female ; Humans ; Male ; Middle Aged ; Parotid Neoplasms ; diagnosis ; pathology ; surgery ; Retrospective Studies

Objective To explore the quality of life (QOL) outcome in patients with allergic rhinitis (AR). Methods A prospective trial was conducted to survey the QOL status of 101 AR patients, in contrast to that of 121 healthy individuals and 97 chronic pharyngitis (CP) patients by generic questionnaire medical outcomes study short-form 36-items health survey (SF-36), and to survey the most troublesome problems of AR patients by disease-specific questionnaire rhinoconjunctivitis quality of life questionnaire (RQLQ). The correlation between SF-36 and RQLQ had also been analyzed. All the results were analyzed statistically. Results By the assessment of SF-36, the scores of 3 domains ( x ± s, the same as follow, the scores were 78. 02 ± 18. 37, 56. 13 ± 17. 49, 78. 81 ± 16. 47, respectively) of AR patients were less than those (84.00 ± 18.36, 74.69±14.13, 83.78±14.31) of healthy individuals (P<0.05), and the scores of 7 domains (the scores were: 91.78 ± 11.78,79.16 ± 30. 23, 78.02 ± 18.37, 56.13 ± 17.49, 78. 81 ± 16. 47, 67. 66 ± 39. 57, 68. 78 ± 13. 65, respectively) of AR patients were similar with those (94.12±6.88, 80.67 ±32.38, 73.57 ±17.96, 59.73 ±16.58, 80.41 ±17.01, 63.58 ±39.99, 66.43 ±13.71) of CP patients (P>0.05). By the assessment of RQLQ, in AR patients, both the nasal symptoms and the practical problems got the highest scores (the scores were 2. 70 ± 1. 29, 2. 53 ± 1. 37 respectively). According to the assessment of the correlation between SF-36 and RQLQ, the correlation was weak (r = - 0. 199～ - 0. 526,P < 0.05). Conclusions The QOL of AR patients decreased compared with that of healthy individuals, but similar with that of CP patients. The most troublesome problems in AR patients were nasal symptoms and the practical problems. Both SF-36 and RQLQ were suitable for assessing the health status of AR patients. SF-36 and RQLQ each covered a different part of the QOL of AR patients, and the combination of the two questionnaires could improve the QOL measurement

Objective To investigate the expression of peripheral blood γδ T cells/CD4 + CD25 + regulatory T cells(Treg) and cytokines interleukin 17 (IL-17) and transforming growth factor β1 (TGF-β1) in patients with allergic rhinitis.Methods From March 2012 to July 2012,32 patients with allergic rhinitis (AR group) and 20 healthy control subjects (control group) were collected.The expression of peripheral blood γδ T cells/Treg cells were measured by flow cytometry and the levels of IL-17 and TGF-β1 were evaluated by ELISA.SPSS 16.0 software was used to analyze the data.Results The percentages of γδT cells in AR group were (13.30 ±8.62)％,which was significantly higher (t =5.18,P ＜0.01) than those in control group (5.18 ± 1.86)％.The percentages of Treg cells in AR group were (1.75 ±0.56)％,which were significantly lower (t =7.46,P ＜ 0.01) than those in control group (4.76 ± 1.74) ％.The IL-17 levels in AR group were (668.55 ±45.15) pg/ml,which were also significantly higher (t =8.97,P ＜ 0.01) than those in control group (573.53 ± 17.42) pg/ml.The TGF-β1 levels in AR group were (0.34 ±0.04) pg/ml,which were also significantly lower (t =9.51,P ＜0.01) than those in control group(0.49 ± 0.06) pg/ml.There was a negative correlation between the percentages of γδ T cells and Treg cells (r =-0.561,P ＜0.01).There was a negative correlation between the percentages of γδ T cells and TGF-β1 levels (r =-0.622,P ＜ 0.01).A positive correlation was shown between the percentages of βδ T cells and IL-17 levels in AR (r =0.469,P ＜ 0.01).A positive correlation was shown between the percentages of Treg cells and TGF-β1 levels in AR (r =0.738,P ＜0.01).There was no correlation between IL-17 levels and the percentages of Treg cells or TGF-β1 levels (r value was-0.111,-0.196,all P ＞ 0.05).Conclusion There are imbalances of γδ T and Treg cells in peripheral blood of patients with allergic rhinitis.γδ T cells may be the main cell to produce IL-17,which may play an important role in allergic rhinitis.