After freeze-drying, the ultrastructure of boar sperms was observed by optical and electron microscopy. The in vitro development ability of the sperm was also examined with intracytoplasmic sperm injection (ICSI). The rate of male pronuclear formation was (68.52%), for cleavage (59.17%) and for blastocyst formation (19.16%) of the trehalose group (0.2 mol/L), significantly higher than those of the 50 mmol/L EDTA group (64.59%, 56.26% and 15.62%) and the control group (35.36%, 52.33% and 8.60%) (P < 0.05). After storage for 60, 120 and 180 d at 4 degrees C, no significant difference in the in vitro development was observed (P > 0.05). The male pronuclear, cleavage and blastocyst formation after ICSI with freeze-dried spermatozoa incubated for 1 h was superior than those incubated for 2 h (P < 0.05). No significant differences in the structures after stored at 4 degrees C or -20 degrees C (P > 0.05) were observed between the trehalose group and EDTA group. The percent of B grade freeze-dried boar spermatozoa in the trehalose group was higher than that of the EDTA group (P < 0.05). Based on the ultrastructure observation, main cryogenic damage in freeze-dried boar spermatozoa was swelling, damage or rupture in the sperm acrosome, neck and tail.
Animals ; Freeze Drying ; Male ; Semen Preservation ; methods ; veterinary ; Sperm Injections, Intracytoplasmic ; veterinary ; Spermatozoa ; Swine ; Trehalose ; pharmacology

Objective To investigate the effect of capillary electrophoresis﹣based multiplex PCR ( CEMP) in detecting pathogens for children respiratory tract infection,and to provide scientific basis for clin﹣ical diagnosis and treatment rapidly and accurately. Methods The cases were defined according to the na﹣tional monitoring program of febrile respiratory syndrome during the 12th Five﹣Year Plan,and the samples were collected from nasopharyngeal swabs,bronchoalveolar lavage fluid and sputum of children with respira﹣tory tract infection hospitalized in Changchun Children′s Hospital from January 2017 to February 2018. Multi﹣plex PCR amplification was performed by one﹣step method, then PCR products were separated by DNA length size with capillary electrophoresis and pathogens were analyzed by"Genemapper software" software. Detecting pathogens included Influenza A virus (InfA),Human Adenovirus (HADV),Boca virus ( Boca), Human Rhinovirus ( HRV), Novel InfA﹣09H1 ( InfA﹣09H1 ) and Seasonal Influenza virus H3N2 ( InfA﹣H3N2),Parainfluenza virus ( HPIV),Human metapneumonia virus ( HMPV), Influenza B virus ( InfB), Mycoplasma pneumoniae (Mp),Chlamydia pneumoniae ( CP),Human Coronavirus ( HCOV),Human Re﹣spiratory Syncytial virus (HRSV). Results The effective detection rate of the CEMP assay was 95. 71%. The positive detection rate of respiratory tract pathogens was 62. 84% and the mixed infection rate was 9. 61%. The mixed infection was mainly InfA and HRSV. The highest three positive rates were named InfA, HRSV and Mp. The positive rate of HRSV was significantly higher in the 0﹣3 age group than that in older group. Different pathogens were detected in different age groups,and the high﹣occurrence season of respiratory tract infection with virus was from December to March of the next year. InfA﹣09H1 was the main prevalent influenza virus in January,February and March 2017,InfA﹣H3N2 was the main prevalent influenza virus in November and December 2017,and the outbreak of InfB was happened in Changchun in late 2017 and early 2018. HRSV was detected only in the coldest season in Changchun from November to March of the next year. Different pathogens were detected in different respiratory infection. HRSV was the main pathogen detec﹣ted in pneumonia; InfA﹣03H2 and HPIV were the main pathogens detected in acute bronchitis; HRV and InfA were the main pathogens detected in upper respiratory tract infection. Conclusion CEMP is an effi﹣cient,rapid and accurate method for the detection of pathogens in patients with respiratory tract infections,and it will have a broad application prospect to develop reagents suitable for clinical diagnosis.

Objective To observe the value of transesophageal echocardiography(TEE)for guiding left atrial appendage closure(LAAC)with LAmbre occluder.Methods Data of 40 non-valvular atrial fibrillation(NVAF)patients who underwent LA AC with LAmbre occluder were retrospectively analyzed.CT angiography(CTA)before treatment,TEE and digital subtraction angiography(DSA)findings during LAAC were comparatively observed,and the correlations of the anchor area diameter and left atrial appendage opening diameter measured with the above three as well as occluder size were analyzed,and TEE and DSA for evaluating peri-device leak(PDL)were compared.Results LAAC were successfully performed with LAmbre occlude in all 40 cases.The diameter of the fixed umbrella was positively correlated with anchor area diameter measured with CTA,TEE and DSA(r=0.79,0.82,0.91,all P＜0.01),of occlusion umbrella was positively correlated with left atrial appendage opening diameter measured with CTA,TEE and DSA(r=0.56,0.89,0.86,all P＜0.01).Immediately after the release of occluder in LAAC,PDL occurred in 16 cases and were detected with both TEE and DSA,while in the rest 24 cases no PDL was found with neither TEE nor DSA.Conclusion TEE had comparable value to DSA for guiding LAAC using LAmbre occluder.

Objective: To investigate the effect of capillary electrophoresis-based multiplex PCR (CEMP) in detecting pathogens for children respiratory tract infection, and to provide scientific basis for clinical diagnosis and treatment rapidly and accurately. Methods: The cases were defined according to the national monitoring program of febrile respiratory syndrome during the 12th Five-Year Plan, and the samples were collected from nasopharyngeal swabs, bronchoalveolar lavage fluid and sputum of children with respiratory tract infection hospitalized in Changchun Children′s Hospital from January 2017 to February 2018.Multiplex PCR amplification was performed by one-step method, then PCR products were separated by DNA length size with capillary electrophoresis and pathogens were analyzed by "Genemapper software" software.Detecting pathogens included Influenza A virus (InfA), Human Adenovirus (HADV), Boca virus (Boca), Human Rhinovirus (HRV), Novel InfA-09H1 (InfA-09H1) and Seasonal Influenza virus H3N2 (InfA-H3N2), Parainfluenza virus (HPIV), Human metapneumonia virus (HMPV), Influenza B virus (InfB), Mycoplasma pneumoniae (Mp), Chlamydia pneumoniae (CP), Human Coronavirus (HCOV), Human Respiratory Syncytial virus (HRSV). Results: The effective detection rate of the CEMP assay was 95.71%.The positive detection rate of respiratory tract pathogens was 62.84% and the mixed infection rate was 9.61%.The mixed infection was mainly InfA and HRSV.The highest three positive rates were named InfA, HRSV and Mp.The positive rate of HRSV was significantly higher in the 0-3 age group than that in older group.Different pathogens were detected in different age groups, and the high-occurrence season of respiratory tract infection with virus was from December to March of the next year.InfA-09H1 was the main prevalent influenza virus in January, February and March 2017, InfA-H3N2 was the main prevalent influenza virus in November and December 2017, and the outbreak of InfB was happened in Changchun in late 2017 and early 2018.HRSV was detected only in the coldest season in Changchun from November to March of the next year.Different pathogens were detected in different respiratory infection.HRSV was the main pathogen detected in pneumonia; InfA-03H2 and HPIV were the main pathogens detected in acute bronchitis; HRV and InfA were the main pathogens detected in upper respiratory tract infection. Conclusion CEMP is an efficient, rapid and accurate method for the detection of pathogens in patients with respiratory tract infections, and it will have a broad application prospect to develop reagents suitable for clinical diagnosis.

Objective: To explore the clinical value of serum autoantibodies and human leukocyte antigen (HLA-B₂₇) molecular testing in Uygur patients with human immunodeficiency virus (HIV) infection. Method: A total of 727 HIV-infected Uygur patients who visited Kuche infectious diseases hospital during May 2016 to March 2017 were include in this study. The other 390 healthy people were enrolled as controls. Serum antinuclear antibodies (ANA), anti-cyclic citrullinated peptide (CCP) antibody, anti-extractable nuclear antigen (ENAs) antibody and HLA-B₂₇ molecule were tested. Result: Among 727 HIV-infected Uygur patients, 317 were males and 410 were females with mean age (35.52±13.44) years old. The mean duration of disease was (6.34±3.05) years. There were 697 (95.87%) patients receiving highly active antiretroviral therapy (HAART) with mean duration of treatment (5.52±3.47) years. The mean CD4⁺T cell count was (520±271) cells/μl in 202 HIV-infected patients, and mean virus load was (108 139±20 498) copies/ml in 20 HIV-infected patients. Rheumatic manifestations were recorded in 238 (32.74%) HIV-infected Uygur patients, mainly with dry mouth and dry eye (15.41%) , alopecia (9.90%) , arthralgia (8.94%) , ect. Compared with the health controls, positive ANA was more common in HIV infected Uygur patients (33.43% vs. 17.43%, P<0.001) with low titers (ANA titer:1∶100) . HIV-infected Uygur patients had higher positive anti-u1-RNP antibodies positive rate (1.10%), but lower anti-SSA antibodies positive rate (0.14%) and anti-CCP antibodies positive rate (0.28%). Patients with positive ANA in HAART group were significantly less than that in non-treatment group (32.71% vs. 50.00%, P=0.049). There were no correlations between ANA and duration of HAART, CD4⁺T cell counts and virus load (r values 0.061, 0.047, 0.121, respectively. P>0.05). Only one female patient was HLA-B₂₇ positive (0.14%), which was significantly lower than that in healthy controls (3.08%) (P<0.001). Also, only one patient was diagnosed with rheumatoid arthritis (RA). Conclusion Autoimmune manifestations are common in HIV-infected Uygur patients. Several autoantibodies are positive, but the coincidence of rheumatic diseases is rare. It′s noted that patients with autoimmune manifestations should be considered as a differential diagnosis of HIV infection.

This study aimed to obtain the first national estimate of the prevalence of autism spectrum disorder (ASD) in Chinese children. We targeted the population of 6 to 12-year-old children for this prevalence study by multistage convenient cluster sampling. The Modified Chinese Autism Spectrum Rating Scale was used for the screening process. Of the target population of 142,086 children, 88.5% (n = 125,806) participated in the study. A total of 363 children were confirmed as having ASD. The observed ASD prevalence rate was 0.29% (95% CI: 0.26%-0.32%) for the overall population. After adjustment for response rates, the estimated number of ASD cases was 867 in the target population sample, thereby achieving an estimated prevalence of 0.70% (95% CI: 0.64%-0.74%). The prevalence was significantly higher in boys than in girls (0.95%; 95% CI: 0.87%-1.02% versus 0.30%; 95% CI: 0.26%-0.34%; P < 0.001). Of the 363 confirmed ASD cases, 43.3% were newly diagnosed, and most of those (90.4%) were attending regular schools, and 68.8% of the children with ASD had at least one neuropsychiatric comorbidity. Our findings provide reliable data on the estimated ASD prevalence and comorbidities in Chinese children.

Radiotherapy is widely used in the management of advanced colorectal cancer (CRC). However, the clinical efficacy is limited by the safe irradiated dose. Sensitizing tumor cells to radiotherapy via interrupting DNA repair is a promising approach to conquering the limitation. The BRCA1-BARD1 complex has been demonstrated to play a critical role in homologous recombination (HR) DSB repair, and its functions may be affected by HERC2 or BAP1. Accumulated evidence illustrates that the ubiquitination-deubiquitination balance is involved in these processes; however, the precise mechanism for the cross-talk among these proteins in HR repair following radiation hasn't been defined. Through activity-based profiling, we identified PT33 as an active entity for HR repair suppression. Subsequently, we revealed that BAP1 serves as a novel molecular target of PT33 via a CRISPR-based deubiquitinase screen. Mechanistically, pharmacological covalent inhibition of BAP1 with PT33 recruits HERC2 to compete with BARD1 for BRCA1 interaction, interrupting HR repair. Consequently, PT33 treatment can substantially enhance the sensitivity of CRC cells to radiotherapy in vitro and in vivo. Overall, these findings provide a mechanistic basis for PT33-induced HR suppression and may guide an effective strategy to improve therapeutic gain.