Objective To investigate distribution of integrain ?1 in human oocytes and embryos before and after maturation/fertilization,dring the first and second meiosis,in morulae and blastocyst stage. Methods Human oocytes and embryos were stained by anti-integrin ?1 (PharMingen 09351D) and FITC-labeled second antibody (PharMingen 110094D),and observed by confocal microscope. Results Integrin ?1 concentrated in the nuclear area in matured oocytes,zygotes and 2-cell stage embryo,indicating that integrin ?1 might involve in mitosis.Dislike mouse,there was no distribution of integrin ?1 on the well-accepted sperm binding area on the oocytes,indicating that integrin ?1 might not involve in the binding and fusion of human sperm to human oocytes.Conclusion It was first postulated that integrin ?1 might actively involve in the pronuclear fusion process in human in vitro fertilized oocytes.The marker of ectotrophoblasts,polarized distribution of integrin ?1, appeared at the morulae stage.The stronger expression in ectotrophoblasts around the inner cell mass might indicate some specific function of the ectotrophoblasts near the ICM.

Objective To investigate the current situation of maternal self- efficacy of breastfeeding and explore its influencing factors in Zhengzhou. Methods Using the general condition questionnaires, breast feeding self-efficacy scale, perceived social support scale and Edinburgh postnatal depression scale to assess the situation of 180 puerperas by the convenience sampling in one obstetric hospital of Zhengzhou. Results The total score of maternal breast feeding self-efficacy was(114.04 ± 21.57)points.The frequency of delivery, the way of delivery and feeding ways, the average income of family were effected with breastfeeding self-efficacy. The total score of social support was(68.87 ± 10.43) points, the total score of puerperas depression was(7.61 ± 4.25) points.The social support score had positive correlation with breastfeeding self-efficacy(r=0.423, P<0.01). Puerperas depression had negative correlation with breastfeeding self- efficacy(r=- 0.342, P<0.01). Conclusions The maternal breastfeeding level in Zhengzhou was at a lower level. The level of maternal breastfeeding self-efficacy scale was impacted by maternal social support degree, the degree of depression. Measures should be taken to enhance maternal social support degree, reduce the degree of depression, in order to improve the level of maternal breastfeeding self-efficacy.

Objective:To express human epididymis protein 4 (HE4) in prokaryotic cells,purify the expressed product and de-termine its activity by immunoassay kit.Methods: The gene encoding HE 4 was cloned using RT-PCR technique from total RNA of ovarian carcinoma cells ES-2,the amplified HE4 gene was cloned into prokaryotic expression vector pGEX-4T-1 and PET26b respective-ly.The recombinant plasmid pGEX-4T-1-HE4 and PET26b-HE4 were constructed and transformed into E.coli BL21 cells respectively, and protein ( GST-HE4 and His-HE4 ) expressions were induced by IPTG and identified by SDS-PAGE and commercial ELISA kit.Results:Restriction analysis and sequencing proved that recombinant plasmid pGEX -4T-1-HE4 and PET26b-HE4 were constructed correctly.The expressed recombinant proteins ,with the relative molecular mass of about 38 000 and 12 000 ,showed specific binding to monoclonal antibody against HE 4.Conclusion:Two kinds of recombinant HE 4 protein are successfully expressed in prokaryotic cells , which laid a foundation of preparation of immunoassay reagents.

AIMTo determine the possible roles of the t-complex testis expressed gene 5 (Tctex5) on sperm functions, the full-length sequence of mRNA was studied and compared in the testis between the normal wild-type and the sterile t-haplotype mutant mice.

METHODSWe applied rapid amplification of cDNA ends, Northern blot and reverse transcription polymerase chain reaction to analyze the full length of Tctex5 mRNAs isolated from testes of the wild-type and the t-haplotype mice. Reverse transcription polymerase chain reaction was used to semi-quantitatively compare expression of Tctex5 transcripts in the 16 tissues and 9.5 day stage embryos in the wild-type mice. E-translation was applied to estimate the amino acid sequences.

RESULTSOne long and one short transcript of Tctex5 mRNA were discovered in mouse testis of wild-type (Tctex5(long-+) and Tctex5(short-+)) and t-haplotype (Tctex5(long-t) and Tctex5(short-t)) mice, respectively. Being enhanced only in the testis, Tctex5(long-t) had 17 point mutations and one 15-bp-deletion in the exon 1 region, comparing with the Tctex5(long-+), whereas the Tctex5(short-t) was similar to the Tctex5(short-+). The short isoforms of Tctex5 mRNAs in the two models encoded exactly the same peptides, but the long isoforms did not. The estimated peptide encoded by Tctex5(long-t) had significant mutations on putative sites of phosphorylation and PP1 binding.

CONCLUSIONWe established that mutations that occur in the Tctex5 long transcript of the t-haplotype mice are important for normal sperm function, whereas the short transcript of Tctex5 might have a conserved function among different tissues.

Animals ; Gene Expression ; Haplotypes ; Infertility, Male ; Male ; Mice ; Microtubule-Associated Proteins ; chemistry ; genetics ; Mutation ; Nuclear Proteins ; chemistry ; genetics ; Protein Phosphatase 1 ; Sequence Analysis, Protein ; Spermatozoa ; metabolism ; Testis ; metabolism ; t-Complex Genome Region

Objective To investigate the clinical value of 2 dimension late Gadolinium enhancement MRI (LGE-MRI) technique for the evaluation of atrial myocardial fibrosis in patients with atrial fibrillation. Methods Forty-nine cases of atrial fibrillation in our hospital from March 2015 to December 2016 were retrospectively collected. The LGE-MR was acquired by the Siemens 3.0 T MR machine before the catheter ablation.The findings of LGE-MR were evaluated by two experienced doctors. The left atrium(LA)were manually segmented into 8 regions in axial view.All patients were classified into 4 stages based on the extent of enhancement, stage 0: absence of enhancement, stage Ⅰ: enhancement appeared in minimal two consecutive slices in single region,stageⅡ:enhancement in two regions,stageⅢ:enhancement in three or more regions. All electroanatomic maps were obtained after electrical conversion during catheter ablation. The Kappa test was used to assess the consistency of LGE-MRI left atrial myocardial fibrosis and CARTO system of the left atrial endocardial voltage reconstruction. Results Forty-nine cases of atrial fibrillation with LGE-MRI and CARTO were included. There were 17 cases of atrial fibrosis stage 0,10 cases of stageⅠ,11 cases of stageⅡ,11 cases of stageⅢaccording to LGE-MRI findings;There were 17 cases of atrial fibrosis stage 0,19 cases of stageⅠ,12 cases of stageⅡ,11 cases of stage Ⅲ with reference to CARTO findings. The diagnostic accuracy of the LGE-MRI atrial fibrosis was 81.6%(40/49),of which the correlation was good(Kappa= 0.751,P<0.001). Conclusions LGE-MRI can accurately assess the degree of left atrial myocardial fibrosis in patients with atrial fibrillation,help to select the proper candidate and strategy in catheter ablation.

Objective:Primary biliary cholangitis (PBC) and Primary Sj?gren′s syndrome (pSS) are autoimmune epithelial inflammatory diseases that share many common clinical symptoms. The aim of this study was to investigate the differences and diagnostic value of Autotaxin (ATX) in PBC and SS.Methods:The clinical data of 237 cases diagnosed with PBC, PBC secondary to SS, pSS and healthy individuals(HC) between September 2020 and September 2021 were retrospectively analyzed. The levels of ATX in each group were measured by enzyme-linked immunosorbent assay (ELISA), and the corresponding sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and area under the curve ( AUC), etc were analyzed. Normally distributed data were expressed as mean ±SD and non-normally distributed as median (IQR). The differences and correlations between ATX and the biochemical tests in each group were assessed by applying the Mann-Whitney U test, Spearman correlation analysis, etc. P<0.05 was considered statistically significant difference. Results:The results showed that ATX was positive in 33.9%, 33.3% and 53.3% for PBC, PBC secondary SS, and pSS, respectively, with the specificities of 93.1%, 100% and 93.2%, respectively. The highest accuracy was achieved in pSS and the sensitivity and specificity were 86.5% and 93.2%, which were higher than those in PBC group(56.8%, 93.1%), respectively. Compared with HC [32.6(21.8, 60.5)ng/ml], ATX levels in PBC[59.3(48.6, 86.3)ng/ml, U=1 750.50, P<0.001], PBC-SS [73.6 (53.3,102.4)ng/ml; U=199.00, P<0.001], and pSS [152.6 (97.4,192.1)ng/ml, U=264.00, P<0.001] were elevated with significant difference ( P<0.05). ATX levels showed a decreasing trend from the pSS group to the HC group. ATX in PBC group[AUC(95% CI)= 0.73(0.651,0.812), P<0.001], PBC secondary SS group [AUC(95% CI)=0.82(0.730, 0.912), P<0.001], and pSS group [AUC(95% CI)=0.94(0.898, 0.984), P<0.001] had prediction accuracy. ATX was associated with total protein ( r=-0.31, P=0.041) level and glutaminase (r=-0.26, P=0.024) level. Conclusion:ATX has diagnostic value in both PBC and SS, and with higher sensitivity and specificity for the latter.

OBJECTIVEThis study aims to analyze the independent risk factors of acute respiratory dysfunction (ARD) in children with airway foreign body and to assess possible prevention and treatment option in the future.

METHODSClinical data of 456 cases of children with airway foreign body were retrospectively collected and analyzed by cluster sampling, including 246 males and 210 females, who received operation in our hospital between July, 2009 and December, 2012, aged 0.5-11 years old, onset to treatment time was 0.15-14 days. Clinical characteristics including age, gender, past medical history, time of onset, temperature, location of the foreign body, category of foreign bodies, complicated by pneumonia, complicated by subcutaneous and mediastinal emphysema were gathered. Temperature, respiratory rate, heart rate, cyanosis, transcutaneous oxygen saturation or arterial blood analysis were assayed before operation. Risk factors with statistical significance were screened with univariate logistic regression analysis, independent risk factors of ARD were determined with multivariate logistic regression analysis.

RESULTSAcute respiratory dysfunction occurred in 78 (17.1%) patients. The foreign bodies in 455 cases were successfully removed brochoscopically in the first time. One case received chest surgery for foreign body removal. Total of 452 cases were successfully extubated and ventilator weaned 4-6 h after brochoscopy. In 2 cases, the ventilator was weaned 2-4 d after brochoscopy in ARD group, and 2 cases with severe pneumonia died. Age, location of the foreign body, temperature, complicated by pneumonia, complicated by subcutaneous and mediastinal emphysema did not show significant difference between acute respiratory dysfunction group and non- acute respiratory dysfunction group (P < 0.05). Multivariate logistic regression analysis showed location of the foreign body and complicated by pneumonia, complicated by subcutaneous and mediastinal emphysema were independent risk factors for ARD.

CONCLUSIONEarly judgement of the risk factors of acute respiratory dysfunction in children with airway foreign body can provide a reference for the operation and perioperation period treatment.

Child ; Female ; Foreign Bodies ; complications ; epidemiology ; Humans ; Male ; Respiratory System ; Respiratory Tract Diseases ; epidemiology ; etiology ; Retrospective Studies ; Risk Factors

Objective:To compare the efficacy of intensity-modulated radiotherapy (IMRT) and three-dimensional conformal radiotherapy (3DCRT) in patients with endometrial cancer, and analyze the influencing factors of prognosis.Methods:The clinical data of 258 patients with stage Ⅰ to ⅢC endometrial cancer undergoing total pelvic radiotherapy from January 2015 to December 2017 in Zhejiang Provincial Cancer Hospital were retrospectively analyzed. Among them, 128 cases were treated with 3DCRT (3DCRT group) and 130 cases were treated with IMRT (IMRT group). Patients with cervical infiltration were given low dose rate vaginal brachytherapy. The local recurrence, disease-free survival (DFS) and overall survival (OS) and adverse reactions were observed. The related factors were analyzed by statistical method.Results:The 258 patients were followed up for 6 to 34 (28.6 ± 1.2) months, and 18 patients had local recurrence, among whom 15 patients were in 3DCRT group and 3 patients were in IMRT group. The local control rates in 3DCRT group and IMRT group were 86.0% (95% CI 74.5 to 92.4) and 94.2% (95% CI 92.8 to 100.0), and there was no statistical difference between 2 groups ( P>0.05). Thirty-six patients had distant metastasis. The distant metastasis rate in 3DCRT group was significantly higher than that in IMRT group: 19.5% (25/128) vs. 8.5% (11/130), and there was statistical difference ( P<0.01). The 2-year DFS rates in 3DCRT group and IMRT group were 81.3% (95% CI 67.6 to 92.4) and 88.2% (95% CI 65.6 to 92.8), and there was no statistical difference between 2 groups ( P>0.05). Cox univariate analysis result showed that age > 68 years, grade 3 malignancy, no lymphadenectomy and > T 1 stage were related to distant tumor metastasis ( P<0.01 or <0.05); Cox multivariate analysis result showed age > 68 years, > T 1 stage and grade 3 malignancy were independent risk factors for distant metastasis in patients with endometrial cancer ( HR = 4.0, 3.3 and 2.1; 95% CI 1.5 to 9.7, 1.5 to 7.4 and 1.2 to 3.5; P<0.01), and no lymphadenectomy was not associated with distant metastasis in patients with endometrial cancer ( P>0.05). The 2-year OS rates in 3DCRT group and IMRT group were 91.6% (95% CI 68.9 to 95.8) and 95.5% (95% CI 65.8 to 98.6), and there was no statistical difference between 2 groups ( P>0.05). The incidences of ≥ 2 grade acute gastrointestinal adverse reactions and grade 2 acute urogenital tract adverse reactions in IMRT group were significantly lower than that in 3DCRT group: 20.0% (26/130) vs. 32.8% (42/128) and 2.3% (3/130) vs. 10.9% (14/128), and there were statistical differences ( P<0.05). One patient in 3DCRT group had late gastrointestinal adverse reaction (grade 3 radiation enteritis). Conclusions:Compared with 3DCRT, IMRT is a safer method for treating patients after endometrial cancer surgery, and the risks of acute gastrointestinal and urogenital tract adverse reactions are lower.

Objective: To explore the effect of ambient fine particulate matters (PM_2.5) exposure on nasal oxidative stress level in patients with allergic rhinitis (AR). Methods: A panel of sixty AR patients was recruited as subjects. Four repeated measurements were carried out from June 2017 to January 2018. Nasal irrigation solution was collected and examined for malondialdehyde (MDA) and superoxide dismutase (SOD). Monitoring data of atmospheric pollutants and meteorological factors during measurement periods were also collected. Time activity pattern combined with micro-environment monitoring method was used to evaluate PM_2.5 exposure. Mixed effect model was applied to analyze the relationship between PM_2.5 exposure concentration and oxidative stress level. Results: 49 subjects accomplished the four repeated measurements, and the mean±SD of their age was (36.7±8.4) years old. The median of MDA and SOD in four measurement periods was 3.70, 3.70, 5.58, 6.24 nmol/ml, and 105.50, 102.50, 95.00, 96.50 U/ml. The concentration of PM_2.5 exposure in four measurement periods was (40.0±2.7), (41.5±2.5), (52.3±5.9) and (74.7±4.9) μg/m³, respectively. Results of single pollutant mixed effect model analysis showed that 0-, 1-, 2-, 3-day lag concentrations of PM_2.5 was significantly positively associated with MDA, about β (95%CI) estimated as 0.24 (0.17, 0.30), 0.34 (0.27, 0.41), 0.32 (0.20, 0.44) and 0.33 (0.23, 0.43), respectively. 0-, 1-, 2-day concentrations of PM_2.5 was significantly negatively associated with SOD, about β (95%CI) estimated as -0.99 (-1.66, -0.31), -1.35 (-2.08, -0.62) and -0.94 (-1.80, -0.07), respectively. Multivariate analysis found that lag 1-day concentration of PM_2.5 was still significantly associated with MDA and SOD after controlling for temperature, age and other influencing factors. For a 10 μg/m³ increase of PM_2.5 concentration, MDA increased 0.26 (95%CI: 0.18, 0.33) nmol/ml, and SOD decreased 0.87 (95%CI: 0.21, 1.53) U/ml. Conclusion Our results suggested that PM_2.5 exposure can aggravate the nasal oxidative stress response of AR patients.