Objective To study the effect of stellate ganglion blocking (SGB) during cardiopulmonary bypass (CPB) on inflammatory reaction.Methods Twenty patients undergoing mitral valve replacement were randomly allocated to 2 matched groups,control group and SGB group (n=10).The control group only received conventional anesthesia,while the SGB group was blocked with 1% lidocaine in addition before the operation for SBG.Blood glucose (BG),Cortisol (Col),IL-1?,IL-6 and TNF-? in the venous blood samples were detected in following 5 time points,20 min before anesthesia,20 min after anesthesia,20 min after operation,20 min afer the start of CPB and 20 min after the end of CPB.Results Col level of the control group (353.09?129.34 ng/ml) was significantly higher than that of the SGB group (486.84?152.20 ng/ml) in 20 min after the end of CPB (P

BACKGROUND: Adipose tissue-derived mesenchymal stem cells (ADMSCs) have the multilineage differentiation potential, and are relatively easier to be obtained, thus they have attracted more and more attention as a new seed cell for cell engineering.OBJECTIVE: To observe the in vitro culture conditions of ADMSCs isolated from rat's subcutaneous adipose tissue, and identify them using immunohistochemical staining.DESIGN: An animal experiment.SETTING; Department of Cardiology, the General Hospital of Chinese PLA.MATERIALS: One healthy male Wistar rat of clean degree, 4 months old, weighing 200 g, was used. DMEM, fetal bovine serum were from GIBCO; Monoclone antibodies of rabbit-anti-rat CD13, CD34, CD44, CD45, CD105, D-related human leucocyte antigen (HLA-DR), factor-Ⅷ, vov Willebrand factor (VWF), Myosin, SABC kits and DAB staining kit from Wuhan Booster Biological Engineering, Co.,Ltd; Adeno-associated virus encoding green fluorescent protein from Vector Gene Technology Company Limited (Beijing).METHODS: The experiments were carried out in the Department of Internal Medicine, the General Hospital of Chinese PLA in October 2006. ① Cell isolation and culture: 0.3 g adipose tissue was cut from subcutaneous adipose tissue of Wistar rat's groin under aseptic condition, then minced and digested before culture, DMEM was changed at 2-3 days after plenty of fusiform-shap ed attached cells were observed under microscope, and the cell growth was observed. The cell concentration was adjusted to 2×107 L-1, then seeded into 96-well plate, and 100 μL for each well. From the second day, 3 wells were randomly selected every day, the cells were released with tripsin, and counted with blood cell counting chamber under inverted microscope. ② Cell viability assay: ADMSCs of passages 3 to 8 were added to DMSO freeze medium, and thawed after 2-4 weeks, and the cell viability was assessed by trypan blue dye exclusion. ③Immunohistochemical staining and identification: 2 ×107 L -1 cells were seeded to culture plate, then the immunohistochemical (SABC method) identification and Oil red O staining were performed to determine the cell surface antibodies of CD13, CD34, CD44, CD45, CD105, HLA-DR, factor-Ⅷ, HLA-DR and VWF. ④Lineage-specific differentiation and identification: The ADMSCs were plated on multi-well chamber and induced with lineage-specific media supplementation at least two weeks and identified by histologic/immunohistochemical assay of Oil red O for adipogenisis, alkaline phosphatase (ALP) stain for osteogenisis and Myosin monoclonal antibody for myogenisis. ⑤Transfected adenovirus carried green fluorescence protein (AD-GFP) medium: The fourth generation of ADMSCs were seeded on 96-well plate, 3 000 cells for each well, serum-free DMEM was changed after 24 hours, and added by AD-GFP at the same time, then transfected with different multiplicity of infection (MOI) of 1∶50, 1∶100, 1∶150 and 1∶200respectively, and then the transfection was observed.MAIN OUTCOME MEASURES: ① Results of cell isolation and culture; ② Cell viability after freezing and thawing; ③Results of immunohistochemical staining and identification; ④ Results of lineage-specific differentiation and identification;⑤ Results of transfected adenovirus carried AD-GFP.RESULTS: ① About 3.6×105 attached cells were obtained from 0.3 g subcutaneous adipose tissue, and these cells could be subcultured for passages in vitro with stable population doubling time. ② The cells were thawed after freezing for 2-3 weeks, and the trypan blue staining showed that the cell viability was above 90%. ③ The immunocytochemical staining showed that CD13, CD44, CD105 were positive and CD45, factor-Ⅷ, HLA-DR and VWF negative in different generations. ④ From the second generation, a few Oil red O positively stained cells were observed, which were obviously increased after prolonging the refreshing. After lineage-specific differentiation, the cells were all positive by Oil red O staining, ALP staining and Myosin immunohistochemical staining. ⑤ 72 hours after transfection, it was observed under fluorescence microscope that most cells were green fluorescence when the MOI value was 1∶200, the transfection was successful, and it was generally determined that the transfection rate was above 90%.CONCLUSION: A large number of ADMSCs with multilineage differentiation potential can be easily obtained from rat adipose tissue, osteoblast, myoblasts, they can be expanded in large quantity and stored in vitro for long time, AD-GFP were also successfully transfected.

Objective To aims Study cardioprotection of different postconditioning algorithm,and investigate the role of MAPK pathway in the process.Methods Sixty SD rats were randomly (random number) divided into 5 groups,sham operation group,reperfusion/injury group (I/R group),gradual decreased reperfusion in postconditioning (GDR,30/10-25/15-15/25-10/30s of reperfusion/re occlusion) group,routine reperfusion in postconditioning (ER,4 cycles of 20/20s of reperfusion/reocclusion) group,and gradual increased reperfusion in postconditioning (GIR,10/30-15/25-25/15-30/10s of reperfusion/re-occlusion) group.Acute myocardial infarction and ischemic postconditioning models were established in the rats.Six hours after reperfusion,3 rats of each group were sacrificed and myocardial tissue were taken out to measure the level of phosphorylation of extracellular signal regulated protein kinase (P-ERK),Phosphorylation of stress-activated protein kinase (stress activated protein kinase,P-JNK),phosphorylation of p38 MAPK (P-p38),tumor necrosis factor-α (TNF-o),cysteine and aspartic protease-8 (Caspase-8) in myocardial tissue and the expression of cytochrome C in the cytosol using Western Blot method.Twenty-four hours after reperfusion all the remaining rats were to measure hemodynamic variables and level of myocardial enzyme,and myocardial tissue were taken out to determine myocardial apoptosis.A one-way analysis of variance (ANOVA) was used,and q tests were employed to determine differences in individual variables existed between groups.Results All three postconditioning modalities were found to provide cardioprotection (P ＜ 0.05) compared with I/R without preconditioning.The GIR provided the best cardioprotection effect,followed by ER and then GDR.Apoptotic index and serum marker levels were reduced far more in GIR than those in ER (P ＜ 0.05).The enhanced cardioprotection provided by GIR was accompanied by significantly increased levels of P-ERK1/2 [(1.82 ± 0.22) vs.(1.54±0.32),P＜0.05],and lower levels of P-p38 [(0.82±0.26) vs.(1.63 ± 0.24)],P-JNK [(0.76±0.28) vs.(1.33±0.21),TNF-o [(0.62±0.20) vs.(1.00±0.12)],Caspase-8 [(0.61 ±0.21) vs.(1.00±0.30)],Cyt-c [(0.66±0.16) vs.(1.68±0.22)] in the cytoplasm (P ＜ 0.05,all compared with ER).In addition,all the variables measured here were significantly improved in the GIR group relative to the GDR group (P ＜ 0.05).Conclusions The method of gradual increased reperfusion in postconditioning could attenuate reperfusion injury more significantly than routine algorithm whereby MAPK pathway played an important role in the process.

Objective To explore the effects of sedationand hemodynamics on different ages patients with the same concentration of dexmedetomidine under general anesthesia.Methods A total of 264 patients (ASAⅠ-Ⅱ)with orthopaedic surgery under general anesthesia in our hospital from April 2013 to May 2015 were divided into 3 groups by age,the young group (group Y,n =76),middle age group (group M, n =107),and old age group (group O,n =81 ).Fifteen minutes before anesthesia,patients were infused dexmedetomidine with 1 μg/kg, maintain the concentration of 0.5 μg·kg-1 ·h-1 and stop at 30 minutes before surgery finished.The SBP、DBP、BIS、HR before anesthesia (T1),pump injection start(T2),tracheal intubation(T3),1 minute after intubation(T4),5 minutes after skin incision(T5),endotracheal ex-tubation(T6)were observed.The dosage of propofoland remifentanil in anesthesia,duration from stop infusion to endotracheal extubation, Ramsay score and adverse reactions 5 minutes after PACU also need to be recorded.Results The level of SBP and DBP were significantly increased at T2,T3 in all groups.Compared with group O,both group Y and group M increased significantly,the difference was statistically significant(P <0.05).The level of SBP and DBP were significantly decreased at T5 in all groups,the difference was statistically significant (P <0.05).There was no significant difference in 3 groups at T4-T6(P >0.05).Compared with T1,the level of HR and BIS were signifi-cantly decreased at T2-T5,the difference was statistically significant(P <0.05).The dosage of propofol and remifentanil in group Y and group Mwas more than that of group O.The extubation time was significantly shorter and the Ramsay score was significantly less than those of the group O,the difference was statistical significance(P <0.05).SAS scores among the three groups was not significant difference (P >0.05). There was no significant difference in the total adverse reaction between group Y and group M(P >0.05),but it was significantly lower than that of group O(P <0.05).Conclusion Dexmedetomidine has good sedative effect in all groups,but older group have more adverse reac-tions and wake up time is extended.The concentratiuon of dexmedetomidine should be adjusted according to the age of patients.

Objective To investigate influence of dexmedetomidine on postoperative cognitive function in elder patients after hip re-placement surgery under spinal anesthesia. Methods Forty elderly patients with ASAⅠ~Ⅲ,undergoing hip replacement with spinal anesth-sia,were randomly divided into dexmedetomidine group( group A) and normal saline group( group B) ,with 20 patients in each group. Dexme-detomidine was given with 1 μg/kg after anesthesia and followed with 0. 5 μg·kg-1 ·h-1 in group A. The equal volume of normal saline was infused in group B. Cognitive function was evaluated before anesthesia,3 and 7 days after surgery by mini-mental state examination( MMSE) . The intraoperative concentration of TNF-α,IL-6,MDA were detected at the time of before surgery(T0),end of surgery(T1),3 days after sur-gery(T2),7 days after suegery. Results There was no significant difference in MMSE score before anesthesia between the two groups (P>0. 05). The difference of MMSE score at postoperative 3 days between two groups was statistical significance (P<0. 05). The MMSE score recovered normal in both groups 7 days later. There was no significant difference of TNF-α,IL-6,MDA concentration at T0 between two groups(P>0. 05). Compared with T0,the concentration of TNF-α,IL-6,MDA at T1,T2 in group B increased,the difference was significant. And the concentration of IL-6 at T1 in group A decreased,compared with that at T0,the difference was significant(P<0. 05). The concentra-tion of TNF-α,IL-6 at T1,T2 and MDA at T2 in group A were lower than those in group B,the difference was significant. (P<0. 05). Con-clusion Dexmedetomidine can decreased the concentration of TNF-α,IL-6,MDA,and improve the postoperative cognitive dysfunction of eld-erly patients who finished the hip replacement surgery under spinal anesthesia.

Objective:To analysis clinical curative effect of laparoscopic radical prostatectomy bladder cancer and influence on serum levels of ferritin (SF),soluble interleukin-2 receptor (SIL-2 R) and rumor specific growth factor (TSGF).Methods:98 cases of bladder cancer who were treated in our hospital from August 2012 to February 2016 were selected and randomly divided into the control group 0=49) and the research group (n=49).The patients in the control group were treated with open radical radical cystectomy,while the patients in the research group were treated with laparoscopic radical cystectomy.Then the operation time,intraoperative blood loss,anal exhaust time,hospitalization,the lymph node cleaning,the serum levels of SF,SIL-2R,TSGF,white blood cells and cortisol,the complications and recurrence rate in the two groups were observed and compared.Results:The operation time of research group was longer than that of the control group,while the intraoperative blood loss,the hospitalization and the anal exhaust time were less than those of the control group,and the differences were statistically significant (P＜0.05).There was no statistically significant difference about the numbers of the lymph node and the recurrence rate between two groups (P＜0.05).After treatment,the serum levels of SF,SIL-2R and TSGF in the two groups decreased,while there was no statistically significant difference between the two groups (P＞0.05);After treatment,the white blood cell count and cortisol rise in the two groups increased,while the research group was lower than that of the control group (P＜0.05).Conclusion:LRC and ORC clinical efficacy similar,both of which can reduce the serum levels of SF,SIL-2R and TSGF of patients with laparoscopic radical prostatectomy bladder cancer.

Objective:To analyze the feasibility of fluorescence staining in the detection of Demodex mites. Methods:A single-center split-face study was conducted, and patients with clinically diagnosed rosacea or seborrheic dermatitis were enrolled from the Department of Dermatology, Southern University of Science and Technology Hospital from October 2020 to June 2021. Samples were obtained from the patients′ cheeks by using the squeeze-adhesion method, and Demodex mites were detected by fluorescence staining and direct microscopic examination separately. The detection rate, number of detected Demodex mites, and time for reading slides were compared between the above two detection methods, and the detection rate and number of detected Demodex mites were further compared between the fluorescence staining and standardized skin surface biopsy (SSSB). Chi-square test was used to compare enumeration data, and paired t-test for measurement data. Results:A total of 433 volunteers aged 28.3 ± 3.5 years were enrolled, including 185 males and 248 females. The performance of fluorescence staining was compared with that of direct microscopic examination in 338 pairs of samples obtained by the squeeze-adhesion method, and compared with that of SSSB in 95 pairs of samples obtained by the squeeze-adhesion method. The detection rate of Demodex mites by fluorescence staining was significantly higher (34.0%, 115/338) than that by direct microscopic examination (31.4%, 106/338; McNemar test, P = 0.004) ; among 118 positive samples, the number of detected Demodex mites by fluorescence staining ([8.0 ± 3.3]/cm 2) was also significantly higher than that by direct microscopic examination ([5.5 ± 2.9]/cm 2, t = 9.21, P < 0.001) ; the time for reading slides undergoing fluorescence staining was significantly shorter (8.3 ± 1.2 minutes) than that undergoing direct microscopic examination (17.3 ± 2.5 minutes, t = 38.44, P < 0.001) ; there was favorable consistency in fluorescence staining results between two clinical laboratorians (kappa value = 0.935, P < 0.001). The detection rate of Demodex mites by fluorescence staining (34.7%, 33/95) was higher than that by SSSB (33.7%, 32/95; McNemar test, P < 0.001) ; among 35 positive samples, the number of detected Demodex mites by fluorescence staining was also significantly higher ([11.4 ± 4.2]/cm 2) than that by SSSB ([9.8 ± 4.8]/cm 2; t = 4.77, P < 0.001) . Conclusion:Compared with direct microscopic examination and SSSB, fluorescence staining was more sensitive in the detection of Demodex mites, with better consistency between different observers and shorter time for reading slides.