Objective:To screen the kinase genes related to radioresistance by high-throughput method and provide a theoretical basis for clinical treatment strategy of oral squamous cell carcinoma.Methods:The tongue squamous carcinoma cell line Cal-27 was infected with lentivirus shRNA kinase library that contains 4 675 different shRNAs regulating 709 human kinase genes. The uninfected cells were removed by puromycin screening. Then, the cells were irradiated with different doses (0, 10, 15 cGy) and continued to culture for 3 d to enrich the differences among groups. Afterwards, the cell genomic DNA was extracted and the complete shRNA sequences were obtained by PCR.Different tags were labeled in shRNAs of each group. An illumina platform was used for next generation sequencing to acquire the shRNA abundance, and then the differentiated expressed genes among different groups were identified.Results:A total of 5 kinase genes (PKLR, IPMK, AURKB, ITPKB and DLG2) were screened out from irradiated cells. Knockdown of PKLR and IPMK increased radiation resistance, while knockdown of AURKB, ITPKB and DLG2 increased radiation sensitivity, and the high expression of these genes would lead to radiotherapy tolerance.Conclusions:In this study, 3 genes relative to radioresistance of oral squamous cell carcinoma were identified by using shRNA lentivirus library combined with next generation sequencing, but the underlying mechanism requires further exploration.

Objective To evaluate the role of silent information regulator 1 (SIRT1) signaling pathway in endotoxin-induced acute lung injury (ALI) in rats.Methods Ninety-six clean-grade healthy male Sprague-Dawley rats,aged 4 weeks,weighing 160-180 g,were divided into 4 groups (n=24 each) using a random number table method:control group (group C),ALI group,ALI + SIRT1 agonist SRT1720 group (group SRT),and ALI + SIRT1 inhibitor EX527 group (group EX).ALI was induced by injecting lipopolysaccharide (LPS) 5 mg/kg (diluted to 0.5 ml with 0.9％ normal saline) in ALI,SRT and EX groups.SRT1720 10 mg/kg was injected via the tail vein,and 30 min later ALI model was established in group SRT.EX527 1 μg/kg was injected via the tail vein,and 30 min later ALI model was established in group EX.Blood samples were collected from the abdominal aorta at 6,24 and 48 h after LPS injection for blood gas analysis,rats were then sacrificed and lungs were removed for determination of wet to dry weight ratio (W/D ratio) and expression of SIRT1,nuclear factor kappa B p65 (NF-κB p65),interleukin-6 (IL-6) and IL-1β protein and mRNA (by Western blot or real-time polymerase chain reaction) and for examination of pathological changes of lung tissues (with a light microscope).Results Compared with groups C,PaO2 was significantly decreased,and PaCO2 and W/D ratio were increased at each time point after LPS injection in ALI,SRT and EX groups,the expression of SIRT1 protein and mRNA was significantly down-regulated,and the expression of NF-κB p65,IL-6 and IL-1β protein and mRNA was upregulated at each time point after LPS injection in ALI and EX groups,and the expression of SIRT1,NFκB p65,IL-6 and IL-1β was significantly up-regulated at each time point after LPS injection in group SRT (P＜0.05).Compared with group ALI,PaO2 was significantly increased,PaCO2 and W/D ratio were decreased,the expression of SIRT1 protein and mRNA was up-regulated,the expression of NF-κB p65,IL-6 and IL-1β protein and mRNA was down-regulated at each time point after LPS injection (P＜0.05),and the pathological changes were significantly attenuated in group SRT,and no significant change was found in the parameters mentioned above at each time point after LPS injection in group EX (P＞0.05).Conclusion Inhibited SIRT1/NF-κB signaling pathway is involved in endotoxin-induced ALI in rats.

Objective To detect the feasibility and safety of applying all-seeing needle in transurethral seminal vesiculoscopy.Methods Retrospective analysis was made with clinical data of 32 patients of hemospermia treated with transurethral seminal vesiculoscopy using all-seeing needle from March 2016 to January 2018.The patients'age was (38.8 ± 8.7) years (27-60 years) and the course of disease was (7.1 ±3.3) months (2-15 months).Ultrasound before operation showed heterogeneous echo,or expansion of the seminal vesicle.MRI showed hemorrhage of the seminal vesicle,or abnormal signal of the seminal vesicle.Patients had levofloxacin or mosisasin anti-infection therapy more than one month and remained uncovered.The operation was performed under subarachnoid anesthesia,and the patients took the lithotomy position.The F4.8 all-seeing needle entered the posterior urethra,the verumontanum was found,and the saline was slowly pushed with a syringe to maintain a clear view.Then,the ejaculatory duct opening was searched on both sides of the verumontanum.If the ejaculatory duct opening cannot be found in the normal position,we entered the needle into the prostatic utricle to find the possible ectopic opening.If the ejaculatory duct opening was still not found,at the 5 and 7 o'clock positions in the prostatic utricle,the needle was probed and punctured into the side wall of the ejaculatory duct.Visible puncture with all-seeing needle can effectively avoid penetrating blood vessels and reduce damage to tissues during puncture.In this study,the ejaculatory duct opening got accessed on the verumontanum in 14 cases,through ectopic openings within the prostatic utricle in 2 cases,and through artificial establishment in 5 and 7 o'clock positions within prostatic utricle in 16 cases.After entering the ejaculatory duct and seminal vesicle,we explored the cavities of the seminal vesicles.For stones or polyps,after replacing the outer sheath to F8,F1.9 stone retrieval basket was applied to remove stones or polyps,followed by rinsing the seminal vesicles with normal saline,0.02％ nitrofurazone,and then 160,000 units of gentamicin into each seminal vesicle.For hemorrhage,after clearing up the blood,seminal vesicles were also washed with normal saline,nitrofurazone,and perfused with gentamicin.In the operation,prostatic utricle stone was found in 5 cases,and seminal vesicle stone was found in 7 cases.One case of seminal vesicle polyp was observed,and in 19 cases,seminal vesicle hemorrhage was seen.Results All the operations were accomplished successfully,the operation time was (55.0 ± 11.3) min (35-82 min).There was no rectal injury or urethral injury during operation.The postoperative catheter was removed at 2 days postoperatively and the patients discharged on the 4th day after surgery.The length of hospital stay was (6.3 ± 0.7) days (5-7 days).One patient had mild hematuria after removal of the urethral catheter and got improved spontaneously;one case developed epididymitis,which improved after anti-infective treatment.6 cases made stone analysis,with 5 cases of magnesium ammonium phosphate hexahydrate,and 1 case of calcium oxalate dihydrate,calcium oxalate monohydrate and carbonate apatite mixed stones.One case of polyp was diagnosed by pathologists to be an inflammatory polyp.Follow-up was performed at 4,6,8,and 12 weeks after surgery.Hemospermia was cured in 24 cases,relieved in the other 8 cases at 8-week follow-up,who received oral antibiotic treatment afterwards.At the 12-week follow-up,the rest 6 cases in 8 got hemospermia recovered,with 2 cases still uncovered.Conclusions All-seeing needle is user-friendly and safe in transurethral seminal vesiculoscopy,with reliable short-term efficacy.

Objective To evaluate the effect of precision anesthesia strategy on postoperative cognitive function in elderly patients undergoing hip replacement.Methods Seventy elderly patients of both sexes,aged 65-85 yr,weighing 50-75 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective unilateral hip replacement under general anesthesia,with years of education＞6 yr,were divided into 2 groups (n =35 each) using a random number table:precision anesthesia group (group P) and routine anesthesia group (group R).Anesthetic protocol and perioperative management were optimized using precision anesthesia strategy in group P.Routine anesthetic protocol and perioperative management were performed in group R.Peripheral venous blood samples were collected at 1 day before operation (T0) and 1,6,12 and 24 h after operation (T1-4) for determination of serum S100β protein,neuronspecific enolase (NSE),interleukin-1beta (IL-1β),IL-6,tumor necrosis factor-alpha (TNF-α) and Creactive protein (CRP) concentrations by enzyme-linked immunosorbent assay.The patient's cognitive function was assessed using Mini-Mental State Examination (MMSE) at T0 and 3 and 7 days after operation (T5,6).Results Compared with the baseline at T0,the serum S100β protein and NSE concentrations were significantly increased at T1-3,the serum IL-1β,IL-6 and TNF-α concentrations were increased at T1-4,the CRP concentrations were increased at T2,3,MMSE scores were decreased at T5 in group R,and the serum S100β protein,IL-1β and IL-6 concentrations were significantly increased at T1-3,the serum NSE and CRP concentrations were increased at T2,the serum TNF-α concentrations were increased at T1-4,and MMSE cores were decreased at T5 in group P (P＜0.05).Compared with group R,the serum S100β protein and IL-1β concentrations were significantly decreased at T1,2,the serum NSE and TNF-α concentrations were decreased at T1-3,the serum IL-6 concentrations were decreased at T2,3,and MMSE scores were increased at T5 in group P (P＜ 0.05).Conclusion Precision anesthesia strategy can improve postoperative cognitive function in elderly patients undergoing hip replacement,which is related to inhibiting inflammatory responses.

Objective To evalute the combination of dexmedetomidine and fentanyl in sedation during awake nasotracheal fiberoptic intubation. Methods One hundred and twenty ASAⅠ or Ⅱ patients scheduled to receive general anesthesia were randomly divided into 3 groups (n = 40 in each group). Patients in group L received an infusion of 1 μg/kg dexmedetomidine,patients in group H received an infusion of 2 μg/kg dexmedeto-midine ,and patients in group DF received an infusion of 1 μ g/kg dexmedetomidine added to 1 μ g/kg fentanyl. Nasotracheal intubation was performed after complete topical anesthesia. HR and MAP were recorded before anes-thesia(baseline,T0),before intubation(T1)and immediately after intubation(T2),respectively. The intubation score(vocal cord movement,coughing and limb movement),fiberoptic intubation score,nasotracheal intubation score and airway obstraction score were assessed in all aptients. On the first post-operative day,recall,adverse events and satisfaction score were also assessed. Results HR and MAP at T1 in three groups were significantly lower than those at T0(P < 0.05,respectively ). HR and MAP at T2 in group L were significantly higher than those in group H and DF(P<0.05,respectively). More incidence of vocal cord closed,severe cough,severe limb movement,heavy grimacing,defensive movement of head and hands after nasotracheal intubation were observed in group L than those in the other two groups. The incidence of airway obstraction and bradycardia in group H were higher than those in group L and DF. Patients in group L had lower postoperative satisfaction scores. Conclusion Adding 1 μg/kg fentanyl to 1 μg/kg dexmedetomidine is a good method for awake nasotracheal fiberoptic intuba-tion,which can prevent the risk of airway obstruction associated with the increase of dexmedetomidine dose,with the achievement of the same favorable sedation.

Objective To evaluate the role of p38 MAPK signal transduction pathway in dexmedetomidine against neurotoxicity induced by bupivacaine.Methods Seventy-two adult male SD rats,successfully implanted with intrathecal catheter without complications,were randomly divided into 6 groups: control group (group C);p38MAPK inhibitor group(group SB);dexmedetomidine group (group D);bupivacaine group (group B);dexmedetomidine and bupivacaine group (group DB);p38MAPK inhibitor and bupivacaine group (group SBB).DMSO 20 μl were injected intrathecally in group C;p38MAPK inhibitor 30 μg and 5% bupivacaine were respectively injected intrathecally in group SB and B;group DB and SBB were respectivel pretreated with dexmedetomidine 75 μg/kg intraperitoneally and p38MAPK inhibitor 30 μg intrathecal injection 20 min before intrathecally injected 5% bupivacaine.Dexmedetomidine 75 μg/kg was injected intraperitoneally in group D.Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured before intrathecal catheter was implanted (T0),before intrathecal administration (T1) and at 4,8 and 12 h and on 1,2,3,4,5 and 6 days after intrathecal administration (T2-T10).At 24 h after intrathecal administration,6 rats were randomly chosen from each group and sacrificed.The lumbar segment (L4-5) of the spinal cord was removed for detecting neuronal apoptosis (by TUNEL) and phosporylated p38MAPK(p-p38MAPK) expression (by Western blot).Results Compared with T0,MWT was significantly increased and TWL was prolonged at T2-T9 in group B,MWT at T2-T7 was significantly increased and TWL at T2-T6 was prolonged in group DB,MWT was significantly increased and TWL was prolonged at T2-T5 in group SBB (P<0.05).Compared with group C,no significant difference was found in MWT,TWL,the apoptotic index and expression of p-p38MAPK in groups D and SB.MWT was significantly increased and TWL was prolonged at T2-T9 in group B,the apoptotic index and expression of p-p38MAPK were significantly increased in group B (P<0.05).Compared with group B,MWT and TWL at T2-T9,the apoptotic index and expression of p-p38MAPK were significantly decreased in groups DB and SBB (P<0.05).Conclusion Dexmedetomidine can inhibit spinal neurotoxicity induced by bupivacaine in rats via inhibiting apoptosis in spinal cord,and inhibition of p38 MAPK signal transduction pathway may be involved in the underlying mechanism.

Objective To discuss the effects and related mechanisms of rapamycin preconditioning on lung injury induced by limb ischemia-reperfusion (IR) in rats.Methods Sixty healthy male SD rats, aged 4-5 months, weighing 250-300 g, were randomly divided into 5 groups (n=12 each) using a random number table: sham operation group (group S);limb ischemia-reperfusion group (group IR);rapamycin 1, 5, 10 mg/kg pretreatment groups (groups R1, R5 and R10).Ischemia-reperfusion of limb was produced by occlusion of bilateral femoral arteries for 2 h followed by 3 h reperfusion.Blood samples were collected to determine serum superoxide dismutase (SOD), malondialdehyde (MDA), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations,1ungs were removed for microscopic examination and for determination of wet/dry lung weight ratio.Results The activity of SOD in groups IR, R1 and R5 was significantly lower than that in group S (P＜0.05).The activity of SOD in groups R1, R5 and R10 was significantly higher than that in group IR, that in groups R5 and R10 was significantly higher than that in group R1, that in group R10 was significantly higher than that in the group R5 (P＜0.05).Serum MDA, IL-1β, IL-6, TNF-α concentrations and wet/dry lung weight ratio were significantly increased in groups IR, R1 and R5 (P＜0.05).Serum MDA, IL-1β, IL-6,TNF-α concentrations and wet/dry lung weight ratio were lower in groups R1, R5 and R10, those in groups R5 and R10 were significantly lower than those in group R1, those in group R10 was significantly lower than those in group R5 (P＜0.05).Compared with group S, the lung tissue injured more significantly in group IR.Compared with group IR, the lung tissue injury gradually reduced in groups R1,R5 and R10.Conclusion Rapamycin pretreatment can reduce lung injury caused by limb ischemia-reperfusion injury in rats in a dose-dependent manner, the greater the dose, the stronger the effect of reducing lung injury caused by limb ischemia-reperfusion injury.The mechanisms may involve attenuating oxidative stress and inhibiting inflammatory response.

Objective To investigate the role of Rho/Rock signaling pathways in ventilator-in-duced lung injury in rats.Methods Ninety-six male SD rats,weighing 300-350 g,were equally and randomly divided into four groups using a random number table (n =24 each):control group (group C),fasudil group (group F),high tidal volume group (group H)and high tidal volume + fasudil group (group HF).Rats in group C and group F received no mechanical ventilation,rats in group H and group HF were intubated and mechanically ventilated (VT = 40 ml/kg,RR = 40 beats per minutes,FiO 2 =40%)for 4 h.The animals in group F and group HF were given intraperitoneal in-jection of fasudil 10 mg/kg at the time 1 h before mechanically ventilated.Six rats were chosen in each group at the time before ventilation (T0 )and at 4,8,24 h after ventilation (T1-T3 ),and blood sam-ples were taken for determination of the levels of serum tumor necrosis factor-α (TNF-α),IL-6 and IL-10,lungs were removed,bronchoalveolar lavage fluid (BALF)was collected to examine protein content,wet/drying (W/D)ratio was determined,which were then stained with haematoxylin and e-osin and examined under microscope,the pathological changes of lungs were scored.Myeloperoxidase (MPO)activity in lung tissue was determined by spectrophptometry.Protein and gene expression of RhoA and Rock2 in the lung tissue were detected by Western blot and reverse transcription PCR (RT-PCR).Results Compared with group C,the serum TNF-α,IL-6 and IL-10,BALF protein content, W/D ratio,the pathological scores,MPO activity,the expression of RhoA,Rock2 protein and mR-NA were significantly increased in group H and HF at T1-T3 (P <0.05 ).Compared with group H, the serum TNF-αand IL-6,BALF protein content,W/D ratio,the pathological scores,MPO activi-ty,the expression of RhoA,Rock2 protein and mRNA were significantly decreased,and the serum IL-10 was significantly increased in group HF at T1-T3 (P <0.05).Conclusion Fasudil can attenuate ventilator-induced lung injury in rats,and the mechanism may be associated with inhibition of the Rho/Rock signaling pathways reducing the inflammatory response.

Objective To investigate the effects of Saikosaponin A (SSA)on cognitive function and cAMP/CREB signaling pathway and expression of BDNF in mice after traumatic brain injury. Methods Sixty SD male mice were randomized into three groups:shame operation group (group S, n =20),trauma group (group T,n =20)and SSA treatment group (group A,n =20).Mice received an administration of SSA 5 mg/kg (group A)or equal volume saline (group S,group T)immediately and once daily for 5 consecutive days after trauma.The cognitive function was detected by Morris wa-ter maze test on day 1,3,7 and 14 after trauma.The hippocampal tissues were harvested after be-havioral tests and homogenized for measuring the levels of brain derived neurophic factor (BDNF)and cyclic AMP (cAMP)by ELISA as well as the levels of cAMP-response element binding protein (CREB)and phosphorylation-cAMP-response element binding protein (pCREB)by western bolt. Results Compared with group S,the escape latency and swimming distance were significantly pro-longed in group T on day 1,3,7 and 14 and group A on day 1,3 after trauma (P <0.05 );while compared with group T,they were significantly shorter in group A on day 7,14 after trauma (P <0.05).Compared with group S,the levels of BDNF,cAMP,CREB and pCREB were significantly de-creased in group T(P < 0.05 ).Compared with group T,the levels of BDNF,cAMP,CREB and pCREB were significantly increased in group A (P <0.05).Conclusion SSA can significantly improve cognitive dysfunction in mice after traumatic brain injury,and the mechanism may be related to the activation of cAMP/CREB signaling pathway and up-regulation of BDNF.

Objective To evaluate the effect of perioperative administration of dexmedetomidine on postoperative ileus after laparocolectomy.Methods Sixty patients scheduled for abdominal surgery were randomly divided into two groups,30 in each group.Group D received dexme-detomidine administeration at a loading dose of 0.6 μg/kg for 10 minutes before induction,followed by an infusion rate of 0.5 μg·kg-1 ·h-1 to 30 min before the end of surgery.The control group re-ceived saline instead of Dex.After the surgery,Group C received intravenous sufentanyl 2 μg/kg, while group D sufentanyl 2 μg/kg combined with Dex 2 μg/kg.Heart rate variability (HRV)were detected before Dex infusion (T0 ),10 minutes after intubation (T1 ), 10 minutes after CO 2 insufflation (T2 ),1 hour after CO 2 insufflation (T3 ),10 minutes after CO 2 desufflation (T4 ),and 10 minutes after extubation(T5 ).The plasma concentrations of epinephrine(E)and norepinephrine (NE)were determined at T0 ,T3 ,T5 ,T7 and T1 0.The recovery of bowel function was evaluated in terms of the first time to fart and intake food.Results Compared with T0 ,HRV of power (TP), high-frequency (HF)power,low-frequency (LF)power and the ratio of LF/HF power were signifi-cantly decreased at T1-T4 in group C and at T1-T5 in group D.The plasma concentrations of E and NE were higher at T3 ,T5 ,T7 and T1 0 in both group C and group D (P <0.05).Compared with T1 ,TP, LF and the ratio of LF/HF were increased at T2-T4 (P <0.05).Compared with group C,TP,LF and the ratio of LF/HF were decreased at T2-T5 ,The plasma concentrations of E and NE were decreased at T3 ,T5 ,T7 and T10 and the time of first flatus was earlier(P <0.05).Conclusion The perioperative ad-ministration of dexmedetomidine during laparocolectomy facilitated the early recovery of bowel func-tion after surgery and decreasede the time of postoperative ileus.