Objective To observe the application effect of bedside contrast-enhanced ultrasound combined with gas-water alternating injection method in severe patients with nasointestinal catheterization.Methods A total of 150 severe patients who were admitted to intensive care unit(ICU)and emergency intensive care unit(EICU)and required nasointestinal catheterization were collected.Patients were separated into the blind insertion method group(catheterization without other auxiliary equipment,n=50),the ultrasound method group(ultrasound-guided catheterization,n=53),and the combined method group(bedside contrast-enhanced ultrasound combined with gas-water alternating injection method,n=47)according to the wishes of patients or their families.The catheterization success rate,feeding standard-reaching rate within one week,catheterization time and incidence of adverse events were compared between three groups.Kappa test was used to compare the consistency of three methods with X-ray examination results after catheterization.Receiver operating characteristic(ROC)curve was used to evaluate the catheterization effect.Results The catheterization success rate and the feeding standard-reaching rate within one week were increased successively in the blind insertion method group,the ultrasound method group and the combined method group,and the catheterization time shortened in turn(P＜0.05).The consistency of blind insertion method,ultrasound method,combined method with X-ray examination was strong(Kappa=0.730),very strong(Kappa=0.835)and very strong(Kappa=0.911),respectively.Results of ROC curve showed that the areas under the ROC curve of the blind insertion method group,the ultrasound method group and the combined method group increased in turn,which were 0.838(95%CI:0.661-1.000),0.918(95%CI:0.763-1.000)and 0.988(95%CI:0.959-1.000),and the combined method group had the best catheterization effect.There were no significant differences in the incidence of adverse events such as hiccup,abdominal distension,diarrhea,aspiration and gastrointestinal bleeding between the three groups(P>0.05).Conclusion Bedside contrast-enhanced ultrasound combined with gas-water alternating injection method can improve the success rate of nasointestinal catheterization in severe patients,shorten the catheterization time and without increasing the incidence of adverse events.

AIM:This study aims to investigate the impact of TWIK-1 channels on abnormal pacemaker activi-ties induced by hypokalemia and to elucidate the underlying mechanisms.METHODS:The gene sequences encoding hu-man TWIK-1,specific TWIK-1 shRNA and TWIK-1-T118i mutant were synthesized and subsequently subcloned into lenti-viral vectors.To knock down the TWIK-1 gene in human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs),the cells were transduced with lentivirus carrying the specific TWIK-1 shRNA sequences.For the overexpression of TWIK-1 or the TWIK-1-T118i mutant in HL-1 mouse cardiomyocytes,the cells received lentiviral transduction containing the respective gene sequences.Patch-clamp techniques were employed to assess the effects of 1 mmol/L extracellular K+on the membrane potentials and whole-cell currents of the cardiomyocytes.RESULTS:Under conditions of 1 mmol/L extra-cellular K+,depolarization of membrane potentials was observed in the hiPSC-CMs and the HL-1 mouse cardiomyocytes ex-pressing human TWIK-1 channel,leading to the induction of abnormal pacemaker activities.This phenomenon could be reversibly abolished by the removal of extracellular Na+or inhibited through TWIK-1 knockdown.In contrast,the mem-brane potentials of HL-1 mouse cardiomyocytes expressing human TWIK-1-T118i mutant hyperpolarized,with no occur-rence of abnormal pacemaker activities.The hiPSC-CMs exhibiting abnormal pacemaker activities at 1 mmol/L extracellu-lar K+demonstrated TWIK-1-like Na+leak currents,which were blocked by quinine,a non-selective blocker of TWIK-1.CONCLUSION:The TWIK-1 channels play a critical role in the development of hypokalemia-induced abnormal pace-maker activities in human cardiomyocytes by facilitating Na+leak currents.

Objective To observe the application effect of bedside contrast-enhanced ultrasound combined with gas-water alternating injection method in severe patients with nasointestinal catheterization.Methods A total of 150 severe patients who were admitted to intensive care unit(ICU)and emergency intensive care unit(EICU)and required nasointestinal catheterization were collected.Patients were separated into the blind insertion method group(catheterization without other auxiliary equipment,n=50),the ultrasound method group(ultrasound-guided catheterization,n=53),and the combined method group(bedside contrast-enhanced ultrasound combined with gas-water alternating injection method,n=47)according to the wishes of patients or their families.The catheterization success rate,feeding standard-reaching rate within one week,catheterization time and incidence of adverse events were compared between three groups.Kappa test was used to compare the consistency of three methods with X-ray examination results after catheterization.Receiver operating characteristic(ROC)curve was used to evaluate the catheterization effect.Results The catheterization success rate and the feeding standard-reaching rate within one week were increased successively in the blind insertion method group,the ultrasound method group and the combined method group,and the catheterization time shortened in turn(P＜0.05).The consistency of blind insertion method,ultrasound method,combined method with X-ray examination was strong(Kappa=0.730),very strong(Kappa=0.835)and very strong(Kappa=0.911),respectively.Results of ROC curve showed that the areas under the ROC curve of the blind insertion method group,the ultrasound method group and the combined method group increased in turn,which were 0.838(95%CI:0.661-1.000),0.918(95%CI:0.763-1.000)and 0.988(95%CI:0.959-1.000),and the combined method group had the best catheterization effect.There were no significant differences in the incidence of adverse events such as hiccup,abdominal distension,diarrhea,aspiration and gastrointestinal bleeding between the three groups(P>0.05).Conclusion Bedside contrast-enhanced ultrasound combined with gas-water alternating injection method can improve the success rate of nasointestinal catheterization in severe patients,shorten the catheterization time and without increasing the incidence of adverse events.

AIM:This study aims to investigate the impact of TWIK-1 channels on abnormal pacemaker activi-ties induced by hypokalemia and to elucidate the underlying mechanisms.METHODS:The gene sequences encoding hu-man TWIK-1,specific TWIK-1 shRNA and TWIK-1-T118i mutant were synthesized and subsequently subcloned into lenti-viral vectors.To knock down the TWIK-1 gene in human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs),the cells were transduced with lentivirus carrying the specific TWIK-1 shRNA sequences.For the overexpression of TWIK-1 or the TWIK-1-T118i mutant in HL-1 mouse cardiomyocytes,the cells received lentiviral transduction containing the respective gene sequences.Patch-clamp techniques were employed to assess the effects of 1 mmol/L extracellular K+on the membrane potentials and whole-cell currents of the cardiomyocytes.RESULTS:Under conditions of 1 mmol/L extra-cellular K+,depolarization of membrane potentials was observed in the hiPSC-CMs and the HL-1 mouse cardiomyocytes ex-pressing human TWIK-1 channel,leading to the induction of abnormal pacemaker activities.This phenomenon could be reversibly abolished by the removal of extracellular Na+or inhibited through TWIK-1 knockdown.In contrast,the mem-brane potentials of HL-1 mouse cardiomyocytes expressing human TWIK-1-T118i mutant hyperpolarized,with no occur-rence of abnormal pacemaker activities.The hiPSC-CMs exhibiting abnormal pacemaker activities at 1 mmol/L extracellu-lar K+demonstrated TWIK-1-like Na+leak currents,which were blocked by quinine,a non-selective blocker of TWIK-1.CONCLUSION:The TWIK-1 channels play a critical role in the development of hypokalemia-induced abnormal pace-maker activities in human cardiomyocytes by facilitating Na+leak currents.

Objective To observe the expression and trends of tight junction proteins Occludin and zonula occlu-den-1(ZO-1)in blood-brain barrier(BBB)of rats with traumatic brain injury(TBI),and to explore the interven-tion effect of cannabidiol(CBD)on the BBB.Methods The TBI model of rat was prepared by modified"Feeney free fall method"and randomly divided into three groups:the sham-operated group(Sham group),the model group(TBI+vehicle group)and the CBD intervention group(TBI+CBD group),with 24 rats in each group.Each group was subdivided into six time points:8 h,1,2,3,5 and 7 d after injury.The expression of Occludin and ZO-1,which are closely related to the permeability of BBB,was detected by immunohistochemistry,immuno-fluorescence staining and Western blot at different points.The permeability of BBB was detected by sodium fluores-cein assay.Results The results of immunohistochemistry showed that compared with Sham group,the positive ex-pression of Occludin and ZO-1 decreased with time after brain trauma(P<0.05),and both reached the lowest level at 2 d.The expression levels of Occludin and ZO-1 were up-regulated after 1 d of CBD intervention(P<0.05).Immunofluorescence staining showed a similar trend to Western blot results,with Occludin and ZO-1 fluo-rescence expression intensity and protein expression reduced after TBI compared with Sham group(P<0.05).And the expression levels of Occludin and ZO-1 were up-regulated after 2 d of CBD intervention(P<0.05).The results of fluorescein sodium experiment showed that the BBB integrity of brain tissue was destroyed after TBI,and the permeability increased after TBI(P<0.01).The permeability of BBB decreased after CBD intervention(P<0.05).Conclusion The expression of tight junction proteins Occludin and ZO-1 decreases after TBI,and the permeability of BBB is disrupted,and CBD intervention reverses the disruption of the BBB by TBI.

Objective To investigate the safety and feasibility of carotid endarterectomy (CEA) combined with carotid artery stent angioplasty (CASA) in treating tandem stenosis of carotid artery. Methods The clinical data of 9 patients with tandem stenosis of carotid artery, who were treated at authors' hospital during the period from January 2013 to October 2014, were retrospectively analyzed. The patients included 7 males and 2 females, with a mean age of (66.0 ±4.2) years. The disease course ranged from 2 months to 36 months, with a mean of 7 months. Clinically, all patients had cerebral ischemia symptoms. Transient ischemia attack was seen in 5 patients and history of cerebral infarction was present in 2 patients. Coronary artery disease was found in 2 patients, hypertension in 6 patients and lower limb ischemia in one patient. After receiving adequate antiplatelet therapy, CEA and CASA were carried out in all patients. Results The technical success rate was 100%, postoperative residual stenosis was less than 30%, no death occurred in perioperative period. After the treatment, the clinical symptoms were improved in all 9 patients;no new stroke or cerebral hemorrhage occurred. After the treatment, 2 patients developed cerebral hyperperfusion-related symptoms such as headache and dizziness, which were much relieved at the time of discharge. The patients were followed up for 4-19 months, with a mean of (10.5±6.2) months. No recurrence of symptoms was observed . In one patient , transcranial Doppler ultrasound performed at 6 months after treatment showed that the carotid artery became moderate restenosis (50%-70%). No death occurred. Conclusion For the treatment of tandem stenosis of carotid artery, CEA combined with CASA is safe and effective, although larger sample and long-term follow-up studies are still needed to further confirm the effect.

Objective To evaluate the POSSUM scoring system as preoperative risk assessment approaches for lower extremity arteriosclerosis obliterans (LEASO).Methods A retrospective study was undertaken in 108 patients ( 120 limbs) diagnosed as long segment LEASO from January 2008 to October 2010,in which,67 patients (74 limbs) receiving percutaneous transluminal stent (PTS) treatment were included into PTS group,and 41 patients (46 limbs) undergoing femoral artery to popliteal artery bypass treatment into bypass group.Rutherford classification was used to evaluate degree of chronic ischemia for lower limb and ankle/brachial index (ABI) for the treatment results in these two groups.The score of POSSUM,physiological score and physiological score without age interference were calculated respectively to estimate the risks for operations in two groups.Results The chronic ischemia conditions for two groups were similar (P =0.543 ).Postoperative follow-up was done for 9 - 15 months,there were no difference between two groups for limb salvage ( P =0.556 ) and patency rate ( P =0.632 ).Risk evaluation for patients: POSSUM score for the PTS group (33 ± 7 ) was similar with that of bypass group ( 32 ± 6 ) ( P =0.369 ) ; Physiological score of POSSUM for PTS group ( 24 ± 7 ) was more than that of bypass group ( 22 ±7) (P =0.033) ; Physiological score without age interference in PTS group (22 ±6)was higher than that of bypass group ( 19 ± 6) (P =0.035).Condusious The physiological score of POSSUM could assess the health status of patients with LEASO,which is more useful for pre-vascular surgery evaluations.

We studied the effects of simulated microgravity on mouse oocytes maturation, and analyzed whether the tail-suspended model can be applied to investigate simulated microgravity effects on reproductive processes in female mice. Mouse oocytes were cultured in vitro with microgravity simulated by a rotating wall vessel bioreactor and by tail-suspended model, and the maturation rate of the mouse oocytes in the two models were examined in vivo. The maturation rate of mouse oocytes cultured in simulated microgravity was 8.93%, and that was 72.33% in 1g gravity. In ratio, oocyte maturation rate had no significant difference between the rotational group and control group. Microgravity simulated by the tail-suspended model inhibited mouse oocytes maturation and increased the rate of oocytes abnormity. The maturation rate of tail-suspended mouse oocytes was 14.54%, which was significantly lower than that of control group. Tail-suspended model should be an ideal model to investigate simulated microgravity effects on reproductive processes of female mice.
Animals ; Cells, Cultured ; Female ; Hindlimb Suspension ; Mice ; Oocytes ; cytology ; physiology ; Oogenesis ; physiology ; Weightlessness Simulation

Objective To investigate the effect and safety of autologons bone margow-mononuclear cell (BM-MNC)transplantation after the bone marrow stimulation in patients with thromboangiitis obliterans (TAO).Methods The bone marrows of 12 patients were stimulated by an injection of the recombinant human granuloeyte-macmphage colony-stimulatory factor(GCSF)for 3-5 days.150-200 ml bone marrow was drown from the iliac spine and the autologous BM-MNC were obtained in each patients.Fifteen lower limbs of 12 patients received implantation of the autologous BM-MNC by an intramuscular iniecdon.A series of subjective indexes(including improvement of pain and cold sensation)and objeetive indexes [including increase of ankle braehial index(ABI),transcutaneons oxygen pressure(TcPO2)and improvement of foot skin ulcer] were used to evaluate the effects.Results The outcomes were evaluated after 2 months of transplantation.The pain-relief rate and the cold feeling improvement rate were 86.7%(13/15)and 93.3%(14/15)respectively.The ABI were 0.38 ±0.05 vs.0.61 ±0.14(P<0.05)before transplantation and 2 months after transplantation respectively.increased in 66.7%(10/15)limbs.The TcPO2 of the ischemic legs increased from(27.47±2.85)mm Hg to(43.53 ±8.38)mm Hg(t=-7.03,P<0.05)after the transplantation,and the improvement rate of TcPO2 was 93.3%(14/15).Skin ulcers in improved in 8/9 limbs.Twelve patients were followed up for all average period of 10 months.The patients'symptoms improved in 80.0%(12/15)limbs,as to the objective index the ABI was0.57±0.13,TcPO2 was(42.07 ±7.81)mm Hg,which improved significandy compared to before treatment(t=-5.33,-7.80,Ps<0.05).skin ulcer healing rate was 66.7%(6/9).The ischemic symptoms in 2 patients were not relieved.There WBS no mortality and high level amputation in all subjects.The complications,such as proliferative retinopathy,malitpmnt tumor,myocardial infarction,stroke or hemangioma were not found in any patients.Conclusion In patients with TAO,intramuscular transplantation of autologous BM-MNC after the bone marrow stimulation has advantages of less bone marrow aspiration,more mononuclear cell content and relatively high safety.It may be a new and effective method to alleviate symptoms and accelerate the healing of skin ulcer.