Objective To investigate the therapeutic effect of three-dimensional interactive reduction of femoral neck and shaft in treatment of irreducible femoral neck fracture.Methods The study enrolled 40 patients with irreducible femoral neck fractures treated by three-dimensional interactive reduction of femoral neck/shaft and internal fixation with three cannulated screws from June 2011 to July 2013 (study group).Frontal and lateral X-ray films were taken after operation.Garden index was used to evaluate the fracture reduction quality and Harris hip score was recorded.Meanwhile,a retrospective analysis was performed on 32 patients with irreducible femoral neck fractures treated by open reduction and internal fixation with cannulated screws between January 2008 and December 2010 (control group).Results Patients in study group obtained satisfactory reduction by minimally traumatic closed reduction.According to the Garden index,fracture reduction quality was level 1 in 24 patients and level 2 in 16 patients.Thirty-two patients were followed up for 12-28 months (mean,21 months),which showed fracture healing.At the final follow-up,Harris hip score was 87 points (range,61-100 points) and 4 patients had femoral head avascular necrosis.By contrast,6 patients in control group showed fracture nonunion.At the final follow-up,Harris hip score was 60 points (range,20-100 points) and 8 patients had femoral head avascular necrosis.Conclusion Three-dimensional interactive reduction of femoral neck and shaft provides good hip function recovery and decreased incidence of femoral head avascular necrosis and fracture disunion compared with the open reduction and internal fixation.

Objective To detect the regional genomic instability of B16 cells treated with 60Co γ-rays by a green fluorescence protein (GFP)-based genomic instability reporting system.Methods Three groups were employed as non-transfection group,vector control group and transfection group.The GFP-marked reporter construct pCMV-EGFP2XhoI for regional genomic instability was successfully transfected into B16 cells using liposome.B16 cells were selected by screening of G418 with a series of concentrations and limiting dilution cultures to yield a single colony.B16 cells with the genomic instability report system were then irradiated by 60Co γ-rays at doses of 0,2 and 4 Gy.The regional genomic instability of B16 cellswas quantified by counting the number of cells with GFP expression.Results B-16 cell strain steadilyexpressing the GFP-based genomic instability reporting system was established successfully.GFP-positiveB16 cells were observed at 1 d after irradiation with 60Co γ-rays at doses of 2 and 4 Gy.Positive correlations between fluorescence intensity and dose and fluorescence intensity and time were also observed.The positive expression rate of GFP followed the increased of dose (F =36.55,36.76,P ＜ 0.05) and time (t =-3.27,-3.16,-4.26,-6.11,-7.17,P ＜ 0.05),and differences between groups were significant.The positive expression rate of GFP increased significantly at 3 d,and maximum expression was observed at 5 d(2.46 ± 0.24 and 3.82 ± 0.35).The level was tending towards stability.Spontaneous GFP expression at a ratio of 1/600 000 was observed in 0 Gy group after 2 weeks of culture.Conclusions The regional genomic instability of B16 cells induced by 60Co γ-rays can be detected using a GFP-labelled genomic instability reporter system.

Objective To construct eukaryotic expressing vector of mouse soluble CD160 and stably transfect into CHO cells for eukaryotic expression.Methods Recombinant soluble CD160(rsCD160) was constructed by gene recombination.Total RNA was extracted from the spleen of C57BL/6 mice.cDNA was amplified for the soluble form of CD160.Then,the PCR product was cloned tO pcDNA3.1 and pEGFP-N1.The recombinant plasmid was identified by restriction map and sequence analy-sis.The soluble CDl 60 expression in CHO cells transfected with recombinant psCDl 60 was verified by RT-PCR and Westernblot.The binding ability of psCD160 tO its ligand was detected by FACS.Results 520 bp mouse soluble CD160 gene was obtained.Recombinant mouse psCD160 was successfully constructed.After transfection,soluble CD160 expression in the culture supernatant of CHO cells was successfully detected.FACS analysis indicated that soluble CD160 could bind tO its ligand.Conclusion Recombinant mouse psCD160 is successfully constructed,which will benefit our further study on soluble CD160 for immune therapy against tumor in the future experiments.

Objective To observe the expression dynamics of lens-related transcription factors in human embryonic stem cell (hESC) differentiated into lentoid body(LB).Methods A "three-stage" protocol was used for LB directional differentiation from hESC in vitro.The hESC (D0) and three differentiation stages cells were collected to analyze the expression dynamics of lens development-associated transcription factors by high throughput RNA sequencing technology in hESC-induced LB.Western blot and cell immunofluorescence were used to observe the involved genes at protein level.Results During D0-D6,cells became more round and compact.And during D7-D18,cells morphology gradually changed to spindle.At the end of D35,three-dimensional and transparent structurelentoid body (LB) was obtained.RT-PCR results showed that the stem cell related genes reduced and the lens specific genes increased significantly,and the LB was characterized by the expression of crystallins.According to clustering analysis of high throughput sequencing,a distinct difference in transcription factors gene expression was observed between D0 and D32.Meanwhile,the difference between D6 and D18 was minimum.The expressions of preplacodal genes,including DLX3,DLX5,DLX6,HES1,HES4,OTX2 and EYA1 increased remarkably at the first induction stage and then decreased.Lens-specification gene SOX2 declined gradually and then increased.In addition,the expression of PAX6 increased during all three induction stages.Furthermore,lens-differentiation genes including MAB21L1,CMAF,PROXI and PITX3 had no significant change in the early induction stage,but increased significantly at the third induction stage.Conclusions The expression dynamics of lens development-associated transcription factors in the hESC induced LB corresponded to those in vivo,which indicate that this induction system can recapitulate early lens development well and lay the foundation of studying lens embryonic development andtranscription factor associated congenital lens diseases.

Objective To assess the impact of short-term, low-dose systemic glucorticosteroids treatment on the clinical outcomes in patients with severe community-acquired pneumonia (SCAP). Methods A multi-center retrospective study was conducted. Data of patients hospitalized with SCAP in five teaching hospitals from Beijing, Shandong and Yunnan Provinces from January 1st, 2013 to December 31st, 2015 were reviewed. Patients were divided into steroids group and non-steroids group according to whether treated with glucorticosteroids during the disease course or not. Data of patients were reviewed, including gender, age, underlying disease, blood routine, biochemical examination and radiology findings (the worst value was recorded if there were more than one value), supportive treatment, complications (hyperglycemia needing insulin treatment and gastrointestinal bleeding) and clinical outcomes [early (0-3 days) treatment failure, late (4-14 days) treatment failure and 30-day mortality, treatment failure was defined as one of the followings: needing noninvasive or invasive ventilation, needing vasopressor use or death]. Univariate and multivariate Logistic regression was performed to evaluate the impact of short-term, low-dose systemic glucorticosteroids on the clinical outcomes in SCAP patients. Results Overall, 3 561 immunocompetent adult and adolescent patients with community-acquired pneumonia (CAP) were screened, 132 SCAP patients were entered into final analysis, including 24 patients in steroids group and 108 patients in non-steroids group. The patients in steroids group were prescribed with methylprednisolone (0.6±0.1) mg·kg-1·d-1 for (4.0±1.7) days. Compared with patients in non-steroids group, patients in steroids group showed younger age [years old: 70.5 (59.0, 75.0) vs. 80.0 (76.0, 85.0)], less frequency of male [41.7% (10/24) vs. 72.2% (78/108)], less comorbidities with cardiovascular [16.7% (4/24) vs. 42.6% (46/108)] and cerebrovascular disease [0% (0/24) vs. 40.7% (44/108)], less confusion [16.7% (4/24) vs. 40.7% (44/108)]; more frequency of chronic obstructive pulmonary disease [COPD, 41.7% (10/24) vs. 13.0% (14/108)], asthma [25.0% (6/24) vs. 1.9% (2/108)], chronic hepatic disease [8.3% (2/24) vs. 0% (0/108)] and respiratory rate≥30 times/min [33.3% (8/24) vs. 9.3% (10/108)] with significant differences (all P < 0.05), the proportion of guideline-based empirical antimicrobial therapy, early needing noninvasive ventilation, late gastrointestinal bleeding, early and late hyperglycemia needing insulin treatment were higher in steroids group than non-steroids group [50.0% (12/24) vs. 21.3% (23/108), 33.3% (8/24) vs. 7.4% (8/108), 20.8% (5/24) vs. 4.6% (5/108), 20.8% (5/24) vs. 1.9% (2/108), 37.5% (9/24) vs. 2.8% (3/108), all P < 0.05]. Adjusted by gender, age, comorbidities and empirical antimicrobial therapy, Logistic regression confirmed short-term, low-dose systemic glucorticosteroids was associated with higher risk for vasopressor usage [odds ratio (OR) = 3.369, 95% confidence interval (95%CI) = 1.369-6.133, P = 0.035], hyperglycaemia needing insulin treatment (OR = 4.738, 95%CI = 1.890-8.652, P = 0.017) in late stage and 30-day mortality (OR = 2.187, 95%CI = 1.265-4.743, P = 0.002). Conclusion Adjunctive treatment with short-term, low-dose systemic glucorticosteroids worsen the clinical outcomes and should not be used to SCAP patients routinely.

Objective:To evaluate the impact of short-term low-medium dose of corticosteroids on the clinical outcomes of patients with community-acquired pneumonia due to influenza A (FluA-CAP).Methods:This was a multicenter, retrospective study, including 693 patients hospitalized with FluA-CAP from Beijing Jishuitan Hospital, Qingdao Municipal Hospital, Beijing Huimin Hospital, Beijing Chao-Yang Hospital and the 2nd People′s Hospital of Yunnan Province during January 1, 2013 to December 31, 2018. The clinical characteristics of patients with or without corticosteroids administration were compared. The first dose of corticosteroids was administrated within 72 hours after admission, with the average dose of methylprednisolone (0.6±0.3) mg/(kg·d) and duration of (4.0±1.2) days. An adjusted logistic regression model was performed to assess the impact of corticosteroids treatment on the clinical outcomes (noninvasive ventilation, invasive ventilation, vasopressor use, admittance to intensive care unit (ICU), 30-day mortality, hyperglycemia needing insulin treatment and gastrointestinal bleeding). Mann-Whitney test and χ2 test were used for the statistical analysis. Results:Among the 693 patients, 132 patients received corticosteroids. Logistic regression analysis revealed that asthma (odd ratios ( OR)=15.528, 95% confidence interval ( CI) 1.953-123.484, P=0.01), chronic obstructive pulmonary disease ( OR=21.904, 95% CI 4.548-105.504, P<0.01) and arterial partial pressure of oxygen (PaO 2)/fraction of inspired oxygen (FiO 2)<300 mmHg (1 mmHg=0.133 kPa, OR=2.701, 95% CI 1.513-4.822, P<0.01) were independent risk factors for corticosteroids use in the FluA-CAP patients. An adjusted logistic regression model showed that low-medium dose corticosteroids administration was associated with decreased risks for early (defined as zero to three days after the first dose of corticosteroids) noninvasive ventilation ( OR=0.342, 95% CI 0.156-0.750, P<0.01), and increased risk for late (defined as four to 14 days after the first dose of corticosteroids) vasopressor use ( OR=2.651, 95% CI 1.913-6.306, P<0.01), late hyperglycemia which needed insulin treatment ( OR=9.739, 95% CI 2.174-21.769, P=0.019), ICU admission ( OR=3.075, 95% CI 1.166-8.143, P<0.01) and the 30-day mortality ( OR=2.372, 95% CI 1.337-4.549, P<0.01). In patients with asthma or chronic obstructive pulmonary disease ( OR=2.343, 95% CI 1.145-4.073, P<0.01) and PaO 2/FiO 2<300 mmHg ( OR=1.961, 95% CI 1.029-4.212, P<0.01), corticosteroids administration increased the risk of 30-day mortality. Conclusion:Low-medium corticosteroids treatment is associated with poor outcomes of FluA-CAP patients, and is not recommended to be used routinely.

Objective To retrospectively analyze the different clinical stages of patients with prostate cancer,and to investigate it's correlation with body mass index (BMI).Methods 363 patients with prostate cancer were enrolled from January 2008 to December 2016.There were 141 cases of stage Ⅱ,Ⅲ in 20 cases,202 cases of stage Ⅳ.According to the stratification of BMI (emaciation group,normal group,overweight group,obesity group),clinical data of different groups of prostate cancer patients were compared to analyze there correlation with BMI.Results Patient's age,pre-PSA concentration,Gleason scores and PSA density were significantly correlated with clinical stage (P ＜ 0.05).Prostate volume and weight had no significant correlation with staging.There was a significant correlation between different strata of BMI and clinical stage (P ＜ 0.05).Conclusion The different strata of BMI are closely related to the clinical stage.The higher BMI,the higher risk of the prostate cancer.

OBJECTIVE: To delineate the clinical and genetic features of a fetus with micrognathia, low-set ears, microtia, polyhydramnios and anechoic stomach by ultrasonography. METHODS: Whole exome sequencing (WES) was carried out to detect genetic variant in the fetus, for which routine chromosomal karyotyping and chromosomal microarray analysis (CMA) yielded no positive finding. Candidate variants were verified by Sanger sequencing and bioinformatic analysis. RESULTS: WES revealed that the fetus has carried a de novo nonsense c.2302C>T (p.Q768X) variant in exon 23 of the EFTUD2 gene, which was detected in neither parent. The variant was unreported previously and may lead to premature termination of the translation of EFTUD2 protein at the 768th amino acid. Bioinformatic analysis predicted the amino acid to be highly conserved and may alter the structure and function of the EFTUD2 protein. CONCLUSION The c.2302C>T variant of the EFTUD2 gene probably underlay the mandibulofacial dysostosis Guion-Almeida type in the fetus. Discovery of the novel variant has enriched variant spectrum of the EFTUD2 gene and provided a basis for genetic counseling and prenatal diagnosis for the family.
Female ; Fetus ; Humans ; Mandibulofacial Dysostosis/genetics* ; Mutation ; Peptide Elongation Factors/genetics* ; Phenotype ; Pregnancy ; Ribonucleoprotein, U5 Small Nuclear/genetics*

Objective:To explore the indications of lung transplantation for pulmonary interstitial fibrosis after COVID-19 and the precautionary measures of medical staff during operation.Methods:Lung transplantation was performed for three cases of COVID-19 from February 15, 2020 to March 1, 2020. The course of disease, mechanical ventilation time, extra-corporeal membrane oxygenation (ECMO) support time, surgical procedures and precautionary measures of medical staff during operation were analyzed.Results:The course of disease were 40, 7, 39 days, the mechanical ventilation time 26, 22, 27 days and ECMO support time 16, 7, 14 days. The postoperative pathological results of three cases indicated pulmonary interstitial fibrosis. One case died on the operative day and two survivors stayed currently in rehabilitation. Thirty-nine medical staff had no symptoms of COVID-19 after 14 days of isolation. The detection of 2019-nCoV nucleic acid was negative for both nasal and pharyngeal swabs.Conclusions:Patients with pulmonary interstitial fibrosis after COVID-19 show no improvement after 1 month of active medical treatment. And the 1-month risk of mortality was over 50% and nucleic acid of 2019-nCoV turned negative. Lung transplantation might be considered. The triple precautionary levels of medical staff during operation prevented the transmission of 2019-nCoV.

OBJECTIVE: To explore the genetic etiology of Vici syndrome in a Chinese family. METHODS: Whole exome sequencing (WES) technology was used to detect gene variants in a fetus of abnormal ultrasonic structure without abnormalities in routine chromosome karyotype analysis and SNP-array. Sanger sequencing and bioinformatics prediction were performed for the suspected variants of the fetus and parents. RESULTS: The fetus and the elder sister have carried c. 2427delC (p.T809fs) and c.1886A>T (p.E629V) compound heterozygous variants of the EPG5 gene, which were respectively inherited from their mother and father. Neither variant was reported previously. According to ACMG guidelines, the c.2427delC variant was predicted as pathogenic, while the c.1886A>T variant was of uncertain significance. PolyPhen-2 and PROVEAN software indicated that c.1886A>T variant was probably damaging. CONCLUSION The c.2427delC and c.1886A>T variants of the EPG5 gene probably underlie the pathogenesis of the Vici syndrome in this family. Above finding has enriched the variational spectrum of EPG5 gene and provided a basis for genetic counseling and prenatal diagnosis for the family.
Aged ; Agenesis of Corpus Callosum ; Autophagy-Related Proteins ; Cataract ; Female ; Humans ; Mutation ; Pregnancy ; Vesicular Transport Proteins/genetics* ; Whole Exome Sequencing