Objective:To evaluate the effect of hydrogen on lipopolysaccharide (LPS)-caused inflammatory responses in BV-2 microglia and the role of autophagy.Methods:The BV-2 microglial cells cultured in vitro were seeded in 6- or 96-well plates and were divided into 4 groups ( n=24 each) using a random number table method: control group (group C), group LPS, hydrogen-rich medium group (group H) and autophagy inhibitor 3-methylpurine group (group 3-MA). In group C, cells were cultured in MEM culture medium supplemented with 15% fetal bovine serum for 24 h. In group LPS, LPS was added at a final concentration of 1 μg/ml, and cells were incubated for 24 h. In group H, LPS was added at a final concentration of 1 μg/ml, the culture medium was replaced with a hydrogen-rich medium at a final concentration of 0.6 mmol/L, and cells were incubated for 24 h. In group 3-MA, 3-methylpurine was added at a final concentration of 2 mmol/L, and the subsequent treatment was similar to those previously described in group H. The cell survival rate was detected by CCK-8 assay.The concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10 and transforming growth factor-β (TGF-β) in supernatant were detected by enzyme-linked immunosorbent assay.The percentage of ionized calcium binding adaptor molecule-1 (Iba-1) +, Iba-1 + CD86 + and Iba-1 + CD206 + cells was detected by flow cytometry.The expression of microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ), LC3Ⅱ, Beclin-1 and p62 was detected by Western blot, and the ratio of LC3Ⅱ/LC3Ⅰ was calculated. Results:There was no significant difference in the cell survival rate among the four groups ( P>0.05). Compared with group C, the concentrations of TNF-α, IL-6, IL-10 and TGF-β and percentage of Iba-1 +, Iba-1 + CD86 + and Iba-1 + CD206 + cells were significantly increased in LPS, H and 3-MA groups, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was significantly down-regulated, and p62 expression was up-regulated in LPS and 3-MA groups, and the ratio of LC3LC3Ⅱ/LC3Ⅰ and Beclin-1 expression was significantly up-regulated, and p62 expression was down-regulated in group H ( P<0.05). Compared with group LPS, the concentrations of TNF-α and IL-6 were significantly decreased, the concentrations of IL-10 and TGF-β were increased, the percentage of Iba-1 + and Iba-1 + CD86 + cells were decreased, the percentage of Iba-1 + CD206 + cells was increased, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was up-regulated, and p62 expression was down-regulated in group H ( P<0.05), and no significant change was found in the above indexes in group 3-MA ( P>0.05). Compared with group H, the concentrations of TNF-α and IL-6 were significantly increased, the concentrations of IL-10 and TGF-β were decreased, the percentage of Iba-1 + and Iba-1 + CD86 + cells was increased, the percentage of Iba-1 + CD206 + cells was decreased, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was down-regulated, and p62 expression was up-regulated in group 3-MA ( P<0.05). Conclusion:The mechanism by which hydrogen reduces LPS-caused inflammatory responses in BV-2 microglia is related to enhancing autophagy and inhibiting microglial activation.

Objective To investigate the relationship between the mechanism underlying hydrogeninduced reduction of sepsis-associated encephalopathy (SAE) and phenotypic transformation of hippocampal microglias in mice.Methods Eighty-eight adult male ICR mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =22 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H2),SAE group and SAE plus hydrogen group (group SAE + H2).Sepsis was induced by cecal ligation and puncture (CLP) in anesthetized mice.Sham and Sham+H2 groups only underwent simple laparotomy.Sham+H2 and SAE+H2 groups inhaled air containing 2％ hydrogen for 1 h starting from 1 and 6 h after CLP,respectively.Mice were sacrificed at 24 h after CLP,and hippocampi were isolated for determination of the levels of tumor necrosis factor (TNF-α),interleukin-6 (IL-6),transforming growth factor-β (TGF-β) and IL-10 (by enzyme-linked immunosorbent assay) and expression of inducible nitric oxide synthase (iNOS) and argininase-1 (Arg-1) (by Western blot).Morris water maze test was performed on 10 mice in each group at days 4-8 after CLP.PResults Compared with group Sham,the levels of TNF-α,IL-6,TGF-β and IL-10 were significantly increased,the expression of iNOS and Arg-1 was up-regulated,the escape latency was prolonged,and the rate of time spent in the target quadrant and the number of crossing the original platform were reduced in SAE and SAE+H2 groups (P＜0.05).Compared with group SAE,the levels of TNF-α and IL-6 were significantly decreased,the expression of iNOS was down-regulated,the expression of TGF-β,IL-10 and Arg-1 was up-regulated,the escape latency was shortened,and the rate of time spent in the target quadrant and the number of crossing the original platform were increased in group SAE+H2 (P＜0.05).Conclusion Hydrogen can promote phenotypic transformation of hippocampal microglias from M1 to M2 and reduce SAE in mice.

Objective To investigate the feasibility and clinical effect of tranexamic acid in prehospital emergency care of patients with craniocerebral trauma.Methods A randomized,placebo controlled trial was carried out on 77 craniocerebral trauma patients [Glasgow Coma Scale (GCS) ≤12 points]enrolled between May 2015 and December 2016.There were 45 males and 32 females,with an average age of 36.5 years (range,19-73 years).Among the patients,37 cases were caused by traffic accidents,19 falling from high places,11 falling when walking,and ten by being hit.According to the random number table method,they were divided into control group (39 cases) and treatment group (38 cases).The treatment group received 1 g of tranexamic acid by intravenous injection within ten minutes on the scene and another 1 g of tranexamic acid within eight hours at the hospital.The control group received 0.9％ isotonic saline.The operation and medication followed the routine process.The arrival time of ambulance and the time of first medication use were recorded.The plasma fibrin degradation products (FDP) and D-dimer at admission and 1 d post-trauma,the percentage of cranitomy operation,case fatality rate,red blood cell transfusion,length of stay in the neurosurgical intensive care unit (NICU),and the Glasgow Outcome Scale (GOS) at day 28 were all recorded and analyzed.Results No significant differences were found between the two groups in gender,age,injury causes,GCS,arrival time of ambulance,and the time of first medication use (P ＞ 0.05).The FDP and D-dimer at admission of the two groups were similar (P ＞ 0.05).One day after admission,the expression of plasma FDP was significantly lower in treatment group than that in control group [6758 (4732,13661) μg/L vs.11740 (8516,21756) μg/L] (P ＜ 0.01).The expression of D-dimer was significantly lower in treatment group than that in control group [1074 (849,1414) μg/L vs.1722 (1389,2330) μg/L] (P ＜ 0.01).Between group differences were insignificant in the percentage of craniotomy operation,case fatality rate,and red blood cell transfusion (P ＞ 0.05).However,treatment group showed shorter stay in NICU [4 (1,12)days vs.2 (0,4)days] and higher GOS [4 (3,5)points vs.5 (4,5)points]than control group (both P ＜ 0.01).Conclusions Tranexamic acid can be applied conveniently in pre-hospital emergency care of craniocerebral trauma patients.It can effectively realize the synchronization of pre hospital transport and treatment,eventually reducing the time of NICU treatment as well as improving the prognosis.

Objective To evaluate midterm outcomes of thoracic endovascular aortic repair (TEVAR) with in situ fenestration (ISF) to revascularize the aortic arch vessels.Methods From Feb 2012 to Dec 2014,10 patients underwent TEVAR with aortic arch vessels revascularized via ISF.There were 6 patients of thoracic aortic aneurysms (TAA) and 4 of type B aortic dissection (TBAD).Patients were followed for all-cause mortality,endoleak of post-TEVAR,integrity and patency of aortic endograft and branch vessels.Results Totally 11 branch vessels [10 left subclavian arteries (LSA),1 left common carotid artery (LCA)] via ISF were revascularized in 10 patients.Patients were followed-up for 24-55 mouths,mean of 42.80 months.1 TAA patient died in 2 years post-TEVAR unrelated to the operation.All fenestrations remained patent,and there were no endoleaks and no occlusion,compression,or fracture of stents.There were no postoperative strokes and left upper limbs ischemia.1 patient had distal aortic endograft pseudoaneurysms formation in 2 years post-TEVAR and underwent reTEVAR treatment.Conclusion Aortic arch vessels revascularization via ISF in TEVAR is safe and feasible.Midterm outcomes is satisfactory.

Objective To establish a cultivating method for obtaining a large number of P0 generation human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Methods The hpcMSCs were isolated from human placental chorion.After primary culturing and culturing for seven days,the culture medium,the non-adherent tissue and the douching normal saline of the primary culture were centrifuged and re-cultured twice.Cell morphology was observed by an inverted microscope.CCK-8 was used to measure the cell growth curve.Flow cytometry was used to detect cell surface markers.Adipogenic and osteogenic differentiation kits were used to assess the cell differentiation potential.Results The obtained hpcMSCs were fibroblast-like adherent cells and (25.54±3.38)×106 cells were obtained per placenta.The total yield of the primary culture,secondary culture and tertiary culture were (11.73±2.09)×106,(11.12±1.42)×106 and (2.69±0.71)×106,respectively,and the incubation time were (12.00±0.64) d,(8.87±0.63) d and (12.33±0.80) d.There was significant differences in incubation time between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the incubation time of the tertiary culture had an increasing trend (P＞0.05).The yield per culture flask of the primary culture,secondary culture and tertiary culture were (1.12±0.15) × 106,(2.10±0.16)×106 and (1.04±0.16)×106,respectively.There was significant differences in the yield per culture flask between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the yield per culture flask of the tertiary culture had a decreasing trend (P＞0.05).There was no difference among the three cultures in the growth curve and the expression of surface markers,and the osteogenic and adipogenic differentiation were all positive.Conclusions The P0 generation hpcMSCs isolated from a choriocarcinoma sample can be doubled by the three cultures compared with the primary culture,which can provide plenty stem cell source for the regenerative medicine.

Objective To determine the median effective concentration (EC50) of lidocaine for obturator nerve block (ONB) guided by a nerve stimulator in patients undergoing transurethral resection of bladder tumor (TURBT).Methods American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients with bladder tumor,scheduled for elective TURBT,required ONB according to the results of cystoscopy or CT examination performed before operation,with body mass index of 19-30 kg/m2,aged 18-64 yr,were enrolled in the study.ONB was performed with lidocaine using the suprainguinal approach under the guidance of a nerve stimulator.The concentration of lidocaine was determined by up-and-down sequential trial.The initial concentration of lidocaine was 1.5％,and the ratio between the two successive concentrations was 1.2.Successful ONB was considered to be positive response.The EC50 and 95％ confidence interval of lidocaine for ONB guided by a nerve stimulator was calculated.Results The EC50 of lidocaine was 0.57％,and the 95％ confidence interval was 0.55％-0.59％ when used for ONB guided by a nerve stimulator.Conclusion The EC50 of lidocaine is 0.57％ when used for ONB guided by a nerve stimulator in the patients undergoing TURBT.

Objective To determine the median effective effect-site concentration (EC50 ) of remifentanil inhibiting responses to laryngeal mask airway insertion when combined with propofol in elderly male patients . Methods Thirty ASA physical status Ⅰ or Ⅱ male patients ,aged 65>yr ,with body mass index <30 kg/m2 , scheduled for elective transurethral resection of bladder tumor or prostate under general anesthesia ,were enrolled in this study .Anesthesia was induced with target-controlled infusion of propofol with a target plasma concentration (Cp) of 3 μg/ml .When Observer′s Assessment of Alertness/Sedation (OAA/S ) score ≤1 ,remifentanil target-controlled infusion was started with the initial target Cp set at 4.0 ng/ml . The concentration of propofol was adjusted until BIS value reached 55-65 ,and then the laryngeal mask airway was inserted .Modified Dixon’s up-and-down method was used to determine the Cp of remifentanil . Each time the Cp of remifentanil increased/decreased in the next patient depending on whether or not the response to laryngeal mask airway insertion occurred . The ratio of the two successive Cps was 1.2 .The response to laryngeal mask airway insertion was defined as development of coughing ,laryngospasm and/or body movement during insertion or within 3 min after insertion .The number of patients in whom inhibition of responses to insertion was effective/ineffective was recorded .The EC50 of remifentanil required to inhibit responses to laryngeal mask airway insertion and the 95% confidence interval when combined with propofol were calculated .Results The EC50 (95% confidence interval ) of remifentanil required to inhibit responses to laryngeal mask airway insertion was 1.86 (1.64-2.12) ng/ml when combined with propofol in elderly male patients .Conclusion The EC50 of remifentanil required to inhibit responses to laryngeal mask airway insertion is 1.86 ng/ml when combined with propofol in elderly male patients .

Objective To evaluate development of gastro-esophageal reflux (GER) during laparoscopic surgery in lateral jack-knife position under general anesthesia through comparing with reverse Trendelenburg/ Trendelenburg position in the patients lying supine.Methods Ninety patients of both sexes,aged 18-64 yr,of ASA physical status Ⅰ or Ⅱ,with body mass index of 18-30 kg/m2,scheduled for elective laparoscopic surgery under general anesthesia,were randomly divided into 3 groups (n =30 each):lateral jack-knife position group (group L),Trendelenburg position group (group T) and reverse Trendelenburg position group (group Tre).Anesthesia was induced with midazolam,sufentanil,propofol and cisatracurium besylate and maintained with propofol and remifentanil given by target-controlled infusion.A pH-sensitive probe was inserted through nose into the lower esophagus and pH value was continuously recorded until 1 min after extubation.GER was defined as pH value ≤ 4 lasting for ≥ 1 min in the lower esophagus during surgery.The development of GER during surgery and the lowest pH value in the lower esophagus when GER developed were recorded.Results Compared with group Tre,the incidence of GER (27％) and total number of times GER had occurred were significantly increased in group L,and no significant changes were found in the indices mentioned above in L and T groups.When GER developed,the lowest pH value in the lower esophagus was 2.1 ± 1.3,2.6 ± 1.2 and 3.5 in L,T and Tre groups,respectively.Conclusion The incidence of GER is 27 ％ during laparoscopic surgery when the patients are in lateral jack-knife position and it is higher than that obtained with reverse Trendelenburg position in the patients lying supine.

Objective To investigate the effect of L-carnitine on the apoptosis in Schwann cells induced by high glucose.Methods The cell line RSC96 cultured in vitro were seeded in 96-well plates at a density of 1.5 × 104/ml (200 μl/well) or in 6-well plates at a density of 2 × 105/ml (2 ml/well) and cultured for 24 h.The cells were randomly divided into 4 groups (n =24 each) using a random number table:normal control group (group C),high glucose group (group H),high glucose + L-carnitine group (group H + L),and mannitol osmotic control group (group M).The cells in group C were incubated in the plain culture medium containing normal glucose (5.6 mmol/L).The cells were incubated in the medium containing glucose 50 mmol/L in group H or in the medium containing glucose 50 mmol/L and L-carnitine 50 μmol/L (final concentration) in group H + L.The cells were incubated in the medium containing normal glucose (5.6 mtmol/L) and mannitol 44.4 mmol/L in group M.At 48 h of incubation,cell growth conditions were observed under inverted microscope,superoxide dismutase (SOD) activity was measured by xanthine oxidase method,malondialdehyde (MDA) content was measured by thiobarbituric acid test,cell viability was measured by MTT assay and cell apoptosis was measured by flow cytometry.The expression of activated caspase-3 and poly (ADP-ribose) polymerase-1 (PARP-1) protein was detected by Western blot.Results Compared with group C,the cell viability and SOD activity were significantly decreased,MDA content and apoptotic rate were increased,and the expression of activated caspase-3 and PARP-1 protein was up-regulated in H and H + L groups,and no significant changes were found in group M.Compared with group H,the cell viability and SOD activity were significantly increased,MDA content and apoptotic rate were decreased,and the expression of activated caspase-3 and PARP-1 protein was down-regulated in group H + L.Conclusion L-camitine can attenuate high glucose-induced apoptosis in Schwann cells by inhibiting oxidative stress responses and down-regulating the expression of activated caspase-3 and PARP-1.

Objective To assess the anesthetic effect of different concentrations of hyperbaric ropivacaine used in lumbar anesthesia for elderly transurethral prostatic resection. Methods We randomly divided 58 elderly patients who un-derwent transurethral prostatic resection with ASA (American Surgical Association) physical status level Ⅰ-Ⅱ into 2 groups as group A and B, using numeric random table method. Each group includes 29 cases. After being punctured between L3-L4, patients were injected with 0.5%(group A) and 0.75%(group B) hyperbaric ropivacaine (diluted with 10%glucose) to the subarachnoid space in a speed of 0.1-0.3 mL/s and the overall dose was 12.5 mg in each case. Then the extents of sen-sory and motor block were compared between two groups, and their anesthesia effect and adverse reactions were also record-ed. Results Compared with group A, onset time of sensory block in group B was significantly shorter (P＜0.05), while onset time for motor block was of no significant difference;By contrast to group A, acting time of sensory block in group B pro-longed significantly (P＜0.05) while acting time of motor block was of no statistically significant difference(P>0.05);By con-trast to group A, the anesthesia effect of group B was more effective while the adverse reactions of the two groups was of no significant difference (P>0.05). Conclusion 0.75%hyperbaric ropivacaine in lumbar anesthesia for elderly patients un-dergoing transurethral prostatic resection could provide a better anesthesia and analgesia effect than 0.5%hyperbaric ropiva-caine without more significant adverse reactions.