AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.

In this study ,we intended to prepare anti‐Toxoplasma gondii aquaporin (TgAQP) peptide antibody which was used to the application in the detection of the aquaporin expression and its subcellular localization of Toxop lasma gondii (T .gondii) ME49 strain .The B cell peptide antigen was designed based on the TgAQP amino acids sequence .After the pep‐tide antigen was conjugated to the KLH ,the fusion antigen was injected into New Zealand rabbits to prepare polyclonal anti‐body ,followed by identification of ELISA ,Western‐blotting and immunofluorescence assays .The ELISA showed that the titer of anti‐TgAQP antibody was about 1∶40 000 .Western blotting revealed the specific affinity of the antigen to polyclonal anti‐body at 29 .9 kDa protein T .gondii .The protein detected by the indirect immunofluorescence assays was distributed in the cy‐toplasm of the parasite .Thus far ,the anti‐TgAQP polyclonal antibody was successfully prepared ,providing a useful tool for further study of biological function and metabolic characteristics of TgAQP .

Objective To evaluate the role of mitochondrial ATP-seusitive potassium (mito-KATP) channels in sevoflurane postconditioning-induced inhibition of oxygen-glucose dcprivation and restoration (OGD/R)-induced pyroptosis in primary rat cardiomyocytes.Methods Cardiomyocytes of newborn Sprague-Dawley rats (＜48 h after birth) were cultured in vitro and seeded in 6-well dishes (2 cm in diameter)or in 96-well plates.The cells were divided into 6 groups (n =15 each) using a random number table:control group (group C),OGD/R group (group O),sevoflurane postconditioning group (group Sev),sevoflurane postconditioning plus 5-hydroxydecanoate (5-HD) group (group SH),5-HD group (group H) and OGD/R plus 5-HD group (group HO).The cardiomyocytes were subjected to oxygen-glucose deprivation for 4 h followed by restoration of oxygen-glucose supply for 24 h.After oxygen-glucose restoration,the cardiomyocytes in the culture media were exposed to 2％ sevoflurane for 1 h to perform sevoflurane postconditioning.At 1 h before oxygen-glucose deprivation,a specific mito-KATP channel blocker 5-HD 100 μmol/L was added to the culture media.Cardiomyocytes were cultured in normal culture atmosphere in group C.Cardiomyocytes were collected at 24 h of oxygen-glucose restoration.Cell pyroptosis was detected by double flow cytometry AlexaFour488 (caspase-1 FLICA staining) and TMR red (DNA staining) staining.The pyroptosis rate was calculated.The cell survival rate was measured by methyl thiazolyl tetrazolium assay.The content of reactive oxygen species (ROS) in mitochondria was determined by 2',7'-dichlorofluorescin diacetate assay.The mitochondrial membrane potential (MMP) was measured by using JC-I fluorescent probe.The expression of interleukin-1beta (IL-1β) was determined by Western blot.Results Compared with group C,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group O (P＜0.05).Compared with group O,the pyroptosis rate and ROS content were significantly decreased,the cell survival rate and MMP were increased,and the expression of IL-1β was down-regulated in group Sev (P＜0.05).Compared with group Sev,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and M MP were decreased,and the expression of IL-1β was up-regulated in group SH (P＜0.05).Compared with group SH,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group H O (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning inhibits OGD/R-induced pyroptosis in primary rat cardiomyocytes is probably associated with increasing mito-KATP channel opening.

Objective To evaluate the role of etomidate post-conditioning on mitochondrial permeability transition pore (mPTP) in the rat cortical neurons subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with Robo receptors.Methods The cortical neurons obtained from Sprague-Dawley rats (＜ 24 h after birth) were cultured in vitro and seeded in 6-well plates (2 ml/well).The neurons were divided into 4 groups (n=24 each) using a random number table:control group (group C),OGD/R group,etomidate post-conditioning group (group E),and etomidate post-conditioning + Robo receptor blocker group (group ER).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In E and ER groups,etomidate was added to the culture medium with the final concentration of 6 μmol/L immediately after onset of O2-glucose supply.In group ER,Robo blocker RoboN was added to the culture medium with the final concentration of 1 μg/ml at 6 h before O2-glucose deprivation.The neuronal apoptosis was detected using Hoechst/PI double staining,the viability of neurons was measured by MTT assay,and the amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.The mitochondria were extracted,and mitochondrial permeability transition pore (mPTP) opening was detected.Results Compared with group C,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in OGD/R,E and ER groups (P＜0.05).Compared with group OGD/R,the apoptosis rate,amount of LDH released,and mPTP opening were significantly decreased,and the cell survival rate was increased in group E,and the apoptosis and amount of LDH released were significantly decreased,and the cell survival rate was increased in group ER (P＜0.05).Compared with group E,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in group ER (P＜0.05).Conclusion Etomidate post-conditioning mitigates OGD/R-induced damage to the cortical neurons through activating Robo receptors and inhibiting mPTP opening in rats.

Objective To study the related factors of no or weak immune response after inoculation of hepatitis B vaccine .Meth‐ods 120casesofadultreceivedinoculationofhepatitisBvaccineinourhospitalwereenrolled,including60casesofnoweakre‐sponse and 60 cases of normal response ,they were divided into non response group and response group .Related factors of no or weakimmuneresponsewereanalyzed.Results (1)singlefactoranalysis:Therewerestatisticalsignificanceofage,gender,BMI, smoking history ,HBV potential infection ,CD4+ T cells and CD8+ T cell content ,IFN‐γand IL‐2 content between two groups ;(2) logistic regression analysis :old age ,male ,smoking history ,high values of BMI ,HPV latent infection ,low content of CD4+ T cells and IFN‐γand IL‐2 ,high content of CD8+ cells were risk factors of no or weak immune response after inoculation of hepatitis B vaccine .Conclusion No or weak immune response after inoculation of hepatitis B vaccine is affected by gender ,age ,smoking ,obesi‐ty ,potential infection of HPV and immune function ,and necessary measure should be taken .

Objective To analyze the risk factors of nosocomial infection in Department of Neurosurgery and to provide evidence for the prevention and treatment of infection.Methods A total of 931 patients with neurosurgery operation in our hospital from January 2012-January 2016 were collected medical history data immediately after admission,including age,gender,underlying diseases,and primary diseases.Surgical records include preoperative white blood cell count,blood glucose level before operation,duration of operation,and reoperation.Hospitalization records include hospitalization time,without the use of corticosteroids,with or without the use of proton there is no pump inhibitor,and tracheal intubation / incision.Patients were divided into infection group and non infection group according to whether the hospital infection occurred during hospitalization.The difference of two groups of clinical data with statistically significant variables was Logistic multivariate regression analysis.Results There were 112 patients with nosocomial infection,the infection rate was 12.03％,and the infection occurred in the postoperative 3-25 d.The main infection site was postoperative wound,accounting for 35.7％;respiratory tract,accounting for 34.8％.There were 64 strains of pathogenic bacteria,81 strains of Gram-negative bacteria,accounting for 64.1％,21 strains of gram positive bacteria,accounting for 32.8％,2 strains of fungi,accounting for 3.1％.There were significant difference between infection group and non infection group in ≥ 60 years,with basic diseases,reoperation,combined with other injuries,white blood cells,abnormal preoperative hyperglycemia,glucocorticoid use,proton pump inhibitors use,tracheotomy,hospitalization time,operation time (P ＜ 0.05).Further Logisitc regression analysis showed that age,reoperation,hospitalization time,preoperative high blood sugar and tracheotomy were the risk factors of nosocomial infection in Department of neurosurgery.Conclusions For the older,reoperation,longer hospitalization time,preoperative hyperglycemia and tracheotomy patients can take specific measures to improve the immunity of the patients,the rational use of antimicrobial drugs to avoid the occurrence of postoperative infection.

Purpose To investigate the prognosis and influencing factor of hematoma complicated from ultrasonography-guided EnCor vacuum-assisted breast biopsy system in minimally invasive surgery for breast lesions, and to provide reference for clinical treatment. Materials and Methods 280 female patients with 486 benign breast lesions underwent minimally invasive excision using a ultrasonography-guided EnCor vacuum-assisted device, occurrence and the inlfuence factors of hematoma complicated from the operation was observed. Results All the 486 lesions were completely excised with EnCor system, hematoma (≥1cm) were found in 47 cases after 24 hours and almost disappeared in 6 months. After analysis of the occurrence of hematoma, it was found that the following situations would lead to a higher incidence of hematoma, which included lumps ≥ 2.5 cm, more than two lesions removed at one time, lumps locating deep inside the areola or the edge of breasts, bigger and less dense breasts, operation in menstrual period, pressure bandaging after operation less than 12 hours and operation without using adrenaline (P<0.05 or P<0.01). Conclusion Benign lesions breast can be effectively excised using ultrasonography-guided EnCor vacuum device, and the factors listed below could affect the occurance of hematoma complicated from the surgery, including the size, location and number of the resected nodules, breast shape, surgery during menstrual period, postoperative pressure bandaging and the use of hemostatics.

Objective To investigate the clinical value of acoustic radiation force impulse (ARFI) imaging in the evaluation of liver functional reserve in patients with hepatic focal lesions. Methods Eighty-six patients with hepatic focal lesions in General Hospital of the People′s Liberation Army from September 2011 to August 2012 were studied. According to Child-Pugh score, 54 of them were classified as grade A, 18 patients were classiifed as grade B, and 14 patients were classiifed as grade C. Seventy patients underwent surgery and 16 patients underwent non-surgical treatment. Shear wave velocity (SWV) values were measured using ARFI technique, and indocyanine green (ICG) clearance tests were also performed to achieve ICG clearance rate constant (ICGK) and ICG retention rate at 15 minutes (ICG15). Correlation between SWV and ICGK, SWV and ICG15 were studied using Pearson correlation, and correlation between SWV and Child-Pugh score were analyzed using Spearman rank correlation. Difference between SWV values of patients underwent surgery and those of patients underwent non-surgical treatment was compared using t-test for independent samples. Analyses of receiver operating characteristic (ROC) curve were performed to investigate the value of ARFI in the evaluation of surgical feasibility for patients with hepatic focal lesion, with the clinical decisions serving as the golden standard. Results There was correlations between SWV and ICGR15, SWV and ICGK, and SWV and Child-Pugh scores (r=0.764,P <0.001;r=-0.686,P=0.000;r=0.864,P=0.000). The SWV of patients who underwent surgery was (2.46±0.45) m/s, and the SWV of patients who underwent non-surgical treatment was (1.54±0.36) m/s, with a signiifcant difference (t=-0.80, P=0.000). The ROC curve analysis demonstrated a highest Youden′s index of 0.775 when the cut-off was 2.06m/s. The corresponding sensitivity was 87.5% and the specificity was 90.0% for ARFI in the diagnosis liver functional reserve deficiency. Conclusion ARFI can effectively evaluate the liver functional reserve in patients with hepatic focal lesions and thus could be beneifcial in the decision of clinical treatment.

Objective To study the clinical value of acoustic radiation force impulse imaging (ARFI) in patients with ischemic type biliary lesion (ITBL) after orthotopic liver transplantation. Methods Between July 2012 to January 2013, forty-one patients in General Hospital of the People′s Liberation Army were enrolled in this study who were admitted for post-transplantation follow-up. Among them, 15 patients were diagnosed as ITBL by ultrasonography. Besides conventional ultrasonography, ARFI was used to detect the shear wave velocity (SWV) of liver tissue at depth of 4 cm and 5 cm respectively. Simultaneously liver function test was performed. Independent-samples t test was performed to compare the difference of SWV between ITBL and non-ITBL group at the same depth. Paired-sample t test was performed to compare the difference of SWV for the same ITBL patient. Pearson correlation analysis was used in analyzing the relation between SWV and liver function. Results The average SWV in depth of 4 cm was (1.561±0.425) m/s and (1.121±0.160) m/s in ITBL and non-ITBL group, respectively. Signiifcant differences were found among the ITBL and non-ITBL groups (t=-3.173, P=0.01). The average SWV in depth of 5 cm was (1.608±0.545) m/s and (1.175±0.173) m/s in ITBL and non-ITBL group, respectively. Signiifcant differences were found among the ITBL and non-ITBL groups (t=-2.454, P=0.034). There was no signiifcant difference between SWV measurements at different depth for the same ITBL patient. For all patients, SWV at different depths were both strongly correlated with alkaline phosphatase (r=0.656, 0.667, respectively;both P=0.000) andγ-glutamyl transpeptidase (r=0.482, P=0.007;r=0.508, P=0.004). Conclusion The liver stiffness measurement is valuable for the clinical evaluation of post-transplantation ITBL.