Objective:To explore the effect of Cfap65 deficiency on mouse spermatogenesis. Methods:CRISPR/Cas9 technology was utilized to construct Cfap65 deficient mice. PCR and Sanger sequencing were adopted to identify mouse genotypes. Mice were divided into Cfap65-/- group ( n=3) and wild-type (WT) group ( n=3) based on genotypes of mice. Fertility test was applied to evaluate the fertility of mice. Sperm morphology of Cfap65 deficient mice was observed by hematoxylin-eosin (HE) staining, immunofluorescence and transmission electron microscope. Real-time fluorescent quantitative PCR was used to detect the expression of Cfap65 mRNA in heart, liver, spleen, lung, kidney and testis tissues of mice. Results:Cfap65 deficient male mice were completely infertile. Compared with wild-type male mice, Cfap65 deficient mice had fewer and less motile epididymal spermatozoa, whose flagellums tend to be short, curled, bent and even absent, and heads tend to be deformed (1.67%±0.44% vs. 33.00%±1.53%), and the differences were statistically significant ( P<0.001). Besides, Cfap65 deficiency led to anomalous structure of manchette of mice. Cfap65 was highly expressed in the testes and lung of adult mice, and the expression of Cfap65 in testes embodied a sharp increase trend from mice aged 4 to 6 weeks (901.90±33.19 vs. 2 144.00±22.92), and the differences were statistically significant ( P<0.001). Conclusion:The expression of Cfap65 is tissue-specific, and the deletion of Cfap65 leads to spermatogenesis failure in male mice, which might be related to the dysfunction of intra-manchette transport.

Objective:To explore the effect of Cfap65 deficiency on mouse spermatogenesis. Methods:CRISPR/Cas9 technology was utilized to construct Cfap65 deficient mice. PCR and Sanger sequencing were adopted to identify mouse genotypes. Mice were divided into Cfap65-/- group ( n=3) and wild-type (WT) group ( n=3) based on genotypes of mice. Fertility test was applied to evaluate the fertility of mice. Sperm morphology of Cfap65 deficient mice was observed by hematoxylin-eosin (HE) staining, immunofluorescence and transmission electron microscope. Real-time fluorescent quantitative PCR was used to detect the expression of Cfap65 mRNA in heart, liver, spleen, lung, kidney and testis tissues of mice. Results:Cfap65 deficient male mice were completely infertile. Compared with wild-type male mice, Cfap65 deficient mice had fewer and less motile epididymal spermatozoa, whose flagellums tend to be short, curled, bent and even absent, and heads tend to be deformed (1.67%±0.44% vs. 33.00%±1.53%), and the differences were statistically significant ( P<0.001). Besides, Cfap65 deficiency led to anomalous structure of manchette of mice. Cfap65 was highly expressed in the testes and lung of adult mice, and the expression of Cfap65 in testes embodied a sharp increase trend from mice aged 4 to 6 weeks (901.90±33.19 vs. 2 144.00±22.92), and the differences were statistically significant ( P<0.001). Conclusion:The expression of Cfap65 is tissue-specific, and the deletion of Cfap65 leads to spermatogenesis failure in male mice, which might be related to the dysfunction of intra-manchette transport.

Objective To explore the factors associated with delayed defecation in long-term ventilated patients in intensivecare unit (ICU) and their potential effect on prognosis.Methods A prospective observational cohort study was conducted. The patients admitted to general ICU of the Affiliated Hospital of Qingdao University from October 1st in 2013 to September 30th in 2015 who underwent mechanical ventilation (MV) for ≥6 days were enrolled, and they were divided into early defecation group (< 6 days) and late defecation group (≥6 days). At admission, clinical nutritional support were given as usual, and gender, age, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score, admission reasons, MV reasons, the usage of morphine and epinephrine/norepinephrine, the highest positive end-expiratory pressure (PEEP), the lowest oxygenation index (PaO2/FiO2) and the lowest systolic blood pressure were collected. Logistic regression analysis was used to analyze the influencing factors of the first defecation time. ICU mortality, the length of ICU stay, central venous catheter (CVC) indwelling time, duration of MV, ICU acquired bacterial infection rate, and the incidence of ventilator associated pneumonia (VAP) were compared between early defecation group and late defecation group. Logisticorgan dysfunction system (LOD) scores and gastric residual volume per day were recorded at the different time of MV.Results Totally 189 patients were enrolled, 39 patients did not satisfied the inclusion criteria and 13 patients gave up treatment or referrals were excluded. Finally 137 patients were enrolled in the analysis, 83 patients in late defecation group and 54 in early defecation group. There were no significant differences in the baseline characteristics such as gender, age, APACHE Ⅱ score, LOD score at 1 day of MV, admission reasons, MV reasons, disgorging and gastric residual volume per day during the first 5 days of MV, enteral nutrition, lactulose treatment in patients with hepatic encephalopathy during the first 5 days of MV, and blood purification treatment between the two groups (allP > 0.05). Compared with the early defecation group, late defecation group had less patients with loose stools or watery stool at first time [15.7% (13/83) vs. 33.3% (18/54)], more patients using morphine and the usage of epinephrine/norepinephrine more than 24 hours [48.2% (40/83) vs. 40.7% (22/54), 42.2%(35/83) vs. 29.6% (16/54)], higher the maximum PEEP level [cmH2O (1 cmH2O = 0.098 kPa): 7.9±3.7 vs. 6.7±3.5], lower the minimal systolic blood pressure [mmHg (1 mmHg = 0.133 kPa): 74.8±28.1 vs. 88.9±30.2] and more severe of hypoxemia [PaO2/FiO2 < 150 mmHg, 54.2% (5/83) vs. 44.4% (24/53)], all of which had significant differences (allP < 0.05). Factors found statistical significances by single factor analysis were enrolled in the multiple regression analysis, which showed that PaO2/FiO2 < 150 mmHg and systolic blood pressure < 90 mmHg were independently associated with a delay in defecation in patients undergoing long-term MV [PaO2/FiO2 < 150 mmHg: adjusted hazard rate: 1.415, 95% confidence interval (95%CI) = 1.061-1.590,P = 0.026; systolic blood pressure 70-89 mmHg:HR = 1.461, 95%CI = 1.164-1.788, P = 0.002; systolic blood pressure ≤ 69 mmHg: adjusted hazard rate= 1.273, 95%CI = 1.010-1.587,P = 0.034). ICU mortality, the length of ICU stay, CVC indwelling time, duration of MV, ICU acquired bacterial infection rate, and the incidence of VAP at 7 days of MV in late defecation group were significantly higher than those of early defecation group [ICU mortality rate: 15.7% (13/83) vs. 7.4% (4/54), the length of ICU stay (day): 17.0 (14.0, 23.0) vs. 15.0 (13.8, 20.0), CVC indwelling time (days): 12.0 (10.0, 14.0) vs. 10.0 (9.0, 11.3), duration of MV (days): 14.0 (10.0, 20.0) vs. 11.0 (9.8, 15.3), ICU acquired bacterial infections rate: 60.2% (50/83) vs. 14.8% (8/54), the incidence of VAP: 32.5% (27/83) vs. 14.8% (8/54); allP < 0.05]. There was no significant difference in LOD score between both groups. The LOD scores at 4 days and 9 days of MV in late defecation group were significantly higher than those of early defecation group (6.41±4.37 vs. 5.21±3.12, 4.33±2.20 vs. 3.50±2.90, bothP < 0.01).Conclusions PaO2/FiO2 < 150 mmHg and systolic blood pressure < 90 mmHg during the first 5 days of MV were independently associated with a delay in defecation in patients undergoing long-term MV. The results suggest that constipation is associated with adverse outcomes in long-term ventilated patients.

Objective To investigate the value of continuous renal replacement therapy (CRRT) coupled with minimally invasive ultrasound-guided percutaneous transhepatic gallbladder drainage (PTGD) for the treatment of severe acute biliary pancreatitis.Methods Hospitalized patients with severe acute biliary pancreatitis were recruited from the intensive care unit (ICU) of the Mfiliated Hospital of Qingdao University from June 2010 to June 2015,and divided into conventional CRRT alone group (n =30) and CRRT + PTGD group (n =30).Comparisons of postoperatively symptoms (time required for abdominal pain relief,time consumed for,gastrointestinal decompression),laboratory findings (WBC,PLT,PCT,CRP,AMS,TBIL,ALT,ALB,Lac) and acute physiology and chronic health evaluation score (APACHE Ⅱ,Balthazar CT,MODS) were carried out between two groups.The occurrence of complications (ARDS,abdominal infection,bile leakage,abdominal hemorrhage,intestinal injury,catheter translocation,catheter dislocation) was observed.The differences in duration of ventilator support,the length of stay in ICU,and fatality rate were compared between the two groups.Results Compared with the conventional CRRT alone group,the postoperative symptoms were significantly relieved,and time required for abdominal pain relief and time consumed for gastrointestinal decompression were evidently shortened in the CRRT + PTGD group (P ＜ 0.05).There were statistically significant differences in laboratory findings (WBC,PLT,PCT,CRP,AMS,TBIL,ALT) between two groups (P ＜ 0.05).The differences in APACHE Ⅱ,Balthazar CT and MODS score between the two groups also presented statistical significance (P ＜ 0.05).The comparisons of the duration of ventilator support [(6.1 ± 1.3) d vs.(9.5 ± 1.4) d] andthe length of stay [(15.7 ± 1.1) dvs.(21.1 ± 2.5) d] between thetwo groups revealed statistical significance (P ＜ 0.05).Conclusions CRRT coupled with PTGD for the treatment of severe acute biliary pancreatitis can effectively eliminate the inflammatory mediators and toxins from patients.On this basis,the coupled therapy with gallbladder puncture and drainage is capable of decompressing the biliary tract,improving liver function,effectively relieving clinical symptoms,minimizing the changes of laboratory findings an,d APACHE Ⅱ score,and thereby optimizing the prognosis of patients.

Objective To investigate the effects of early rehabilitation therapy in mechanically ventilated ICU in patients.Methods A randomized controlled trial was carried out.Sixty mechanically ventilated patients,with tracheal intubation or tracheostomy more than 48 h and less than 72 h,were admitted to the intensive care unit (ICU) of the Affiliated Hospital of Medical College Qingdao University from May 2010 to May 2012.These patients were randomly (random number) divided into two groups,the rehabilitation group and the control group,30 patients in each group.In rehabilitation group,rehabilitation therapy was taken twice daily,and the training time and intensity was adjusted according to the condition of the patients.Early rehabilitation therapy included head up actively,transfer from the supine to sit,out of bed,transfer to a chair,standing bedside bed and walking bedside bed.The patient' s body mass index,days to first out of bed,duration of mechanical ventilation,ICU stay,APACHE Ⅱ score,highest FiO2,lowest PaO2/FiO2 and hospital mortality of patients were compared between rehabilitation group and control group.The differences between the groups were compared using t test.Results There was no significant difference in body mass index,APACHE Ⅱ score,highest FiO2,lowest PaO2/FiO2 and hospital mortality between rehabilitation group and control group (P ＞ 0.05).Patients in the rehabilitation group had shorter first out of bed time [(3.8 ± 1.2) d vs.(14.9 ±4.7) d; P =0.00],duration of mechanical ventilation [(5.6±2.1) dvs.(12.7±4.1) d; P=0.005] andICUstay [(7.3±2.8) dvs.(15.2±4.5) d;P =0.01] compared with control group.Conclusions Early rehabilitation therapy was safe and effective in improving the outcomes of mechanical ventilation patient.