Objective To clone the full length staphylococcal protein A(SPA) gene from Staphylococcus aureus (ATCC6538), and subsequently study the gene structure and antibody binding ability.Methods The full length and the functional region of the SPA gene were cloned into pHisSUMO vector respectively, and expressed in E. coli. The full length and the functional fragment of the SPA protein were detected for antibody binding ability and stability. The functional fragment of the SPA protein fused with SUMO was coupled to the CNBr-activated agarose for antibody purification from rabbit serum. Results A variant of the full length SPA gene was cloned, which has been submitted to GenBank (the accession number is EU695225). Two fusion proteins had the same antibody binding ability as the untagged SPA protein. However, the formers was more stable than the latter at the tested conditions. SUMO-SPA conjugated-agarose kept high efficiency for antibody binding. Conclusion To our knowledge, the full length SPA gene of S.aureus(ATCC6538) is a novel variant. The SUMO tag can improve the stability of the functional region of the SPA protein without damaging the antibody binding ability. This fusion protein has been used for antibody purification successfully.

Insulin is the most common medicine used for diabetic patients, unfortunately, its effective time is short, even the long-acting insulin cannot obtain a satisfactory effect. Fibroblast growth factor (FGF)-21 is a recently discovered glucose mediator and expected to be a potential anti-diabetic drug that does not rely on insulin. In this study, db/db mice were used as the type 2 diabetic model to examine whether mFGF-21 has the long-term blood lowering effect on the animal model. The results showed that mFGF-21 could stably maintain the blood glucose at normal level for a long-term in a dose-dependent manner. Administration of mFGF-21 once a day with three doses (0.125, 0.25 and 0.5 mg x kg(-1)) could maintain blood glucose of the model animals at normal level for at least 24 h. Administration of mFGF-21 every two days with the same doses could maintain blood glucose of the model animals at normal level for at least 48 h, although it took longer time for blood glucose to reach to normal level depending on doses used (twenty injections for 0.125 mg x kg(-1) and 0.25 mg x kg(-1) doses, ten injections for 0.5 mg x kg(-1) dose). Surprisingly, the blood glucose of the treated model animals still maintained at normal level for 24 h after the experiment terminated. Glycosylated hemoglobin level of the animals treated with mFGF-21, which represented long-term glucose status, decreased significantly compared to the control group and the insulin group. The results suggest that FGF-21 has potential to become a long-acting and potent anti-diabetic drug.

Objective:To investigate the effect of the possibility of sleep-disordered breathing (SDB) as assessed by the four-variable score on the platelet function and the risk of stroke recurrence in patients with acute ischemic stroke.Methods:Patients with acute ischemic stroke admitted to the Department of Neurology, Weihai Municipal Hospital from January 2020 to January 2021 were enrolled prospectively. Main inclusion criteria: admission within 24 h of onset; National Institutes of Health Stroke Scale score ≤3; Receiving aspirin + clopidogrel dual antiplatelet therapy. All patients were divided into a high possibility group and a low possibility group of SDB according to the four-variable score. 7±2 d after dual antiplatelet therapy, PL-12 multi-parameter platelet function analyzer was used to detect the maximum aggregation rate (MAR). The patients were followed up for 6 months after discharge and the recurrence of ischemic stroke was observed. The mediating effect model was established with the high possibility of SDB as the independent variable, MAR as the intermediary variable and stroke recurrence as the dependent variable. Firstly, MAR as the dependent variable and high probability of SDB as the independent variable were analyzed by linear regression; then, a binary logistic regression analysis was performed with ischemic stroke recurrence as the dependent variable and the high probability of SDB and MAR as independent variables. Results:A total of 213 patients were enrolled in the study. The average age of the patients was 62.70 ± 10.04 years old. There were 146 male (68.5%) and 121 patients (56.8%) were in the high possibility group (56.8%). During the follow-up period, 24 patients (11.3%) had stroke recurrence. Univariate analysis showed that arachidonic acid (AA) induced MAR (MAR-AA) and adenosine diphosphate (ADP) induced the MAR (MAR-ADP) in the high possibility group of SDB were significantly higher than those in the low possibility group (all P<0.05); MAR-AA and MAR-ADP in the recurrent group were significantly higher than those in the non-recurrent group (all P<0.05), and the proportion of high possibility of SDB in the recurrent group was significantly higher ( P=0.008). Binary logistic regression analysis showed that homocysteine (odds ratio 1.132, 95% confidence interval 1.048-1.223; P=0.002) and having high possibility of SDB (odds ratio 6.351, 95% confidence interval 1.134-35.566; P=0.035) were the independent risk factors for stroke recurrence in patients treated with dual antiplatelet therapy. Intermediary effect analysis showed that MAR had a significant intermediary effect on the risk of stroke recurrence in patients with high probability of SDB. Conclusion:The MAR and stroke recurrence rates in the high possibility group of SDB were significantly higher than those in the low possibility group, and its stroke risk was probably mediated by platelet hyperreactivity.

Objective To investigate the specific vessels signs in lung cancer by 64-slice spiral CT in first phase perfusion ima-ging so as to get more reasonable time about CT angiography .Methods Among the 47 cases ,there were 38 cases of central and 9 cases of peripheral primary lung cancer underwent 64-slice spiral CT in first phase perfusion imaging :one scan was obtained every 1 seconds during 8-38 seconds with 8 section × I without scanning interval after injection .Precontrast and postcontrast attenuation on every leisions was recorded ,to calculate the peak height(PH) and peak height time by time density curves TDC of pulmonary le-sions to aorta ,those reflect the the tumor feeding artery Imaging about lesions of lung cancer and inflammation .observed and calcu-lated the display rate and display time of lesions of lung cancer and inflammation .means of t test was used for statistics .Results a-bout the peak heights of in pulmonary artery and aorta phese ,statistically significant differences were found between inflammation and lung cancer(P<0 .05) .no statistically significant differences were found between central and peripheral lung cancer (P>0 .05) . about the peak heights time in aorta phase ,no statistically significant differences were found among three groups (P>0 .05) .The tumor vascularity were discoved in lesions in patients with lung cancer (44/47 cases ,93 .62% );The erosion narrow pulmonary ar-tery were discoved in central lung cancer (37/38 cases ,97 .37% ) ,in peripheral lung cancer(6/9 cases 66 .67% );no abnormal pulmo-nary artery were discoved only in 4 patients with lung cancer .Both tumor vascularity and abnormal pulmonary artery were most dis-plaied in 18-31s in CT angiography .Conclusion 64-slice spiral CT in first phase dynamic perfusion imaging can order completely show the specific vessels signs in lung cancer and reflect the tumor feeding artery Imaging of lung cancer .Analysising those benefi-ted to select reasonably the time of CT angiography and Improve the rate of lung cancer diagnosis .

Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.

Objective To investigate the effect of surgical treatment on survival in colorectal carcinoma patients with synchronous hepatic metastasis.Methods The retrospective case-control study was done on 953 consecutive patients with synchronous colorectal hepatic metastasesl from January 2003 to December 2013.Results Median survival time (46.7 months)and 5-year survival rate (32％) for patients with resected hepatic metastases was significantly superior to that of with nonoperative treatment (17 months,4％).Expanded criteria for hepatic metastases resection raised resection rates (31％ vs.13.6％,P ＜0.05).For patients with resectable hepatic metastases,the inhospital cost for simultaneous resection group was lower than that in the staged resection group (36 698 vs.45 134 RMB,P ＜ 0.05).For patients of asymptomatic primary tumor with unresectable hepatic metastases,resection of the primary tumor was associated with an improved median survival (18.0 vs.15.0 months,P ＜ 0.05) Conclusions Expanding indications of hepatic metastases resection can improve survival in patients with synchronous colorectal hepatic metastases.Simultaneous resection of primary tumor and hepatic metastases were indicated in patients with resectable synchronous colorectal hepatic metastases.Resection of primary tumor was recommended for asymptomatic patients with unresectable hepatic metastases.

Objective To improve the recognition of Pancreatitis, Panniculitis, and Polyarthritis syndrome (PPP syndrome). Methods We described the diagnosis and treatment of a patient with PPP syndrome, including the clinical, radiological, and pathological features. Results Here we report a 67-year-old man with chronic calcified pancreatitis with multiple subcutaneous nodules and polyarthritis, but without any abdominal pain or other abdominal symptoms. His serum pancreatic amylase and lipase were markedly elevated. Abdomen CT scan showed features of chronic calcified pancreatitis. MR imaging of ankle revealed intramedullary fat necrosis. Biopsy from subcutaneous swellings revealed fat necrosis without vasculitis and typical "ghost-like cells". He failed to response to corticosteroids therapy. When the diagnosis of pancreatitis was confirmed, he was managed conservatively with supportive measures, and recovered. Conclusion The diagnosis of PPP syndrome is often difficult when abdominal symptoms are absent, be aware the association between panniculitis and polyarthritis with pancreatic disease may lead to a prompt diagnosis and management.

Objective To summarize our clinical experience of cryoablation for renal cancer and to analyze the therapeutic indication,security,selection of cryoablation and outcomes.Methods Sixty-four patients suffered with T1a renal cell carcinoma were enrolled in this study from March 2012 to March 2018.Among them,5 cases were senile patients (≥ 80 years),5 cases complicated with other cancers,3 cases complicated with renal insufficiency,4 cases complicated with decompensated cirrhosis,3 cases with bilateral renal cancer,4 cases with solitary kidney cancer and 39 cases with some other complications.The preoperative serum creatinine level was(80.5 ± 38.2)μmol/L.The patients underwent laparoscopic singlesite (LESS) renal cryoablation,conventional laparoscopic renal cryoablation,or percutaneous image-guided cryoablation according to individual situation.Contrast-enhanced CT scan or MRI were used during the procedures and follow-up was performed.Results All operations were completed successfully and technical success was achieved as well in all cases.Blood transfusion was necessary for 2 cases because of hemorrhage.The mean diameter of the mass was (2.6 ± 0.90) cm,the median volume of blood loss was 50ml(10-110 ml),and the mean operation time was(96.0 ± 24.5) min.The median inpatient hospital stay was 3 d (1-6 d).In one case,digital subtraction angiography (DSA) embolization was performed due to hemorrhage after surgery.None of the other cases had intraoperative or postoperative complications.The serum creatinine level after surgery was not significantly decreased [postoperative (83.8 ±42.1) μmol/L,P =0.64].The contrast-enhanced CT or MRI of the kidneys one week postoperatively showed uniform low density in all lesion areas,which represented complete ablation and regression of the tumor.All cases were followed up regularly.One case showed relapse at the 6 th month follow-up and underwent cryoablation again.Another case,who was not regularly followed up,relapsed at 69th month after surgery.No relapse was observed in the other cases during the follow-up.Conclusions Renal cancer cryoablation is a safe,feasible and efficacious therapy for the patients who suffered from unresectable T1a renal cell carcinoma because of high surgical risk or multifocal lesions.