Objective To improve a HPLOUV method for the determination of Tramadol hydrochlo-ride in human plasma. Method Alkalinized samples of plasma were extracted with dichloromethane. Tramadol and internai standard were analyzed on Elite C18 column (200mm?5.0mm, 5?m) with a acetoni-trile-phsphate buffer (pH6.5) (74:26, v/v) mobile phase and detedted at 220nm. Results The Calibra-tion plots in human plasma were linear (r=0.9984, n =5) from 10 to 800 ng/ml. The limit of detection was 10 ng/ml. For 20, 100, 400 ng/ml check samples, intra-run precisions (RSD) were 3.81% -5.44% and inter-run preceisions (RSD) were 3.95% -4.41% , respectively. The average recoveries were 89% , 96% and 92% for the low, middle and high check samples, respectively. Conclusion The improved ana-lytical method for Tramadol hydrochloride was found to be sensitive, simple and rapid, suitable for applica-tion in forensic toxicological analysis and routine determination of numerous samples.

Objective To evaluate the role of mammalian target of rapamycin (mTOR) signaling pathway in dexmedetomidine-induced reduction of renal ischemia-reperfusion (I/R) injury in rats and the relationship with hypoxia-inducible factor 1 (HIF-1α).Methods Seventy-two male Sprague-Dawley rats, aged 10-12 weeks, weighing 220-260 g, were randomly divided into 4 groups (n=18 each) using a random number table: sham operation group (group S), group I/R, dexmedetomidine group (group Dex) ,and rapamicyn + dexmedetomidine group (group Rpm+Dex).Renal I/R was produced by occlusion of bilateral renal pedicles for 35 min follow by reperfusion in anesthetized rats in I/R, Dex and Rpm+Dex groups.Bilateral renal pedicles were only exposed, and then the abdominal cavity was closed in group S.Dexdetomidine 50 μg/kg was injected intraperitoneally at 30 min before I/R in group Dex.In group Rpm+Dex, rapamicyn 1.5 mg/kg and dexdetomidine 50 μg/kg were injected intraperitoneally, and renal I/R model was established 30 min later.Immediately after onset of reperfusion, and at 4 and 24 h of reperfusion (T1-3) , blood samples were collected from the caudal vein for measurement of serum creatinine and blood urea nitrogen (BUN) concentrations.After blood sampling at T1-3, the rats were sacrificed, and the renal specimens were obtained for detection of HIF-1αt, erythropoietin (EPO) and mTOR expression by Western blot.Their kidneys were removed at T3, and cut into sections which were stained with haematoxylin and eosin and examined under microscope.Acute renal tubular necrosis was scored.The cell apoptosis in renal tissues was detected by TUNEL assay, and apoptosis index (AI) was calculated.Results Compared with group S,the concentrations of serum creatinine and BUN, expression of HIF-1α, EPO and mTOR at T2,3 , AI at T3 and acute renal tubular necrosis score were significantly increased in the other three groups (P＜ 0.05).Compared with group I/R, the concentrations of serum creatinine and BUN were significantly decreased, and the expression of HIF-1α, EPO and mTOR was up-regulated at T2,3 , and AI and acute renal tubular necrosis score were decreased in group Dex (P＜0.05) , and no significant change was found in the parameters mentioned above in group Rpm + Dex (P ＞ 0.05).Conclusion The mTOR signaling pathway is involved in dexmedetomidine-induced reduction of renal I/R injury, which may be related to dexmedetomidine-produced up-regulation of HIF-1α expression in renal tissues of rats.

Objective To investigate the protection effect of dexmedetomidine against H2O2 injury in Human renal tubular epithelial cells(HK-2 cells). Methods HK-2 cells cultured in vitro were randomly divided into four groups(n = 24): control group, dexmedetomidine pretreatment group, H2O2 injury group, H2O2 injury +dexmedetomidine pretreatment group. Cell viabilities were measured by MTS assay, cell apoptosis were detected using flow cytometry, and expression of HIF-1α protein was quantified by western blot. HK-2 cells were divided into 8 groups by combining with three treatment factors such as PI3K inhibitor LY294002, dexmedetomidine and H2O2 injury. MTS assay was used to detect cell viability and western blot was used to quantify protein expression of HIF-1α,Bcl-2 and Bax after treatment in each group. Results Dexmedetomidine significantly increased the level of HIF-1α、 Bcl-2 in HK-2 cells after H2O2 injury, thus improved viabilities and reduced apotosis of cells. Moreover, effect on H2O2 injury cells of Dexmedetomidine was reversed by PI3K inhibitor LY294002. Conclusion Dexmedetomidine could protect against H2O2 injury by up-regulating HIF-1α expression through activating PI3K/Akt/mTOR signaling pathway in HK-2 cells.

OBJECTIVE:To evaluate the economics of somatostatin,ulinastatin,octreotide and gabexate preventing hyperamy-lasemia and pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP). METHODS:Medical records of 316 cho-ledocholithiasis patients underwent ERCP were selected from our hospital during Jul. 2008-Apr. 2016,and then divided into blank control group(58 cases),somatostatin group(64 cases),ulinastatin group(65 cases),octreotide group(68 cases)and gabexate group (61 cases) according to the use of protease inhibitor. Before ERCP,blank control group received routine treatment as fast-ing,parenteral nutrition support,acid suppression,etc. Based on it,other 4 drug groups received prophylactic drug use according to package inserts 6 h before ERCP. The incidence of hyperamylasemia and pancreatitis after ERCP,VAS scores 3,24,48 h after surgery and the occurrence of ADR were compared among 5 groups. The cost-effectiveness analysis was used to evaluate the eco-nomics of therapy plans in each group. RESULTS:Compared with blank control group,hospitalization time of somatostatin group, ulinastatin group,octreotide group and gabexate group were shortened significantly;the incidence of hyperamylasemia and pancre-atitis were significantly decreased;VAS score 3,24,48 h after surgery were significantly decreased,with statistical significance (P<0.05). Compared among 4 groups,above indexes had no significant difference,and the incidence of ADR was in low level and had no statistical significance(P>0.05). The cost-effectiveness ratio of somatostatin group was the lowest and has cost-effec-tiveness advantage. The results were supported by incremental cost-effectiveness and sensitivity analysis. CONCLUSIONS:Soma-tostatin,ulinastatin,octreotide and gabexate can significantly prevent the occurrence of hyperamylasemia and pancreatitis after ER-CP,and relieve pain with good safety. Somatostatin can achieve the best therapeutic efficacy at the lowest cost,so it is the best plan for hyperamylasemia and pancreatitis after ERCP.

Objective:To established a modified implanting model of VX2 liver tumor in rabbit on the base of the classic implanting method, and compared the results within the two methods. Methods:30 rabbits with the mean weight of (2.65?0.29)kg were divided randomly into two groups with 15 rabbits each. The rabbits in Group A received classic implantation for induction of the liver tumor model, and Group B were inducted by injecting a piece of tumor tissue into the left anterior lobes of liver. Implanting time of each group was recorded and compared, and spiral CT scan was performed at 8th day, 15th day, 22nd day, 29th day postoperatively. The manifestation of tumors in CT scan was observed and tumor volume was calculated simultaneously with formula V=1/2ab2 (a=the shortest diameter and b=the longest diameter).Each tumor was confirmed through pathology. Results:The implanting time of Group A and Group B were (9.47?2.85)min and (5.85?1.62)min, respectively, with significant difference between them. Besides, there was statistical difference of the achievement ratio between two groups, as it was 53.3% for Group A and 86.7% for Group B. No significant difference was found for the tumor growth between two groups. Conclusion:Modified implanting method for induction of the rabbit liver tumor model was superior to the classic implanting method.

Objective To investigate the therapeutic effects and mechanism of anti-radiation pneumonia decoction(ARPD) on radiation induced lung fibrosis in rats.Methods One hundred and five male SD rats in a SPF grade were divided into Chinese medicine group,single radiation group and control group by random digits table method,with 35 in each group.After anesthetization,rats in Chinese medicine and single radiation groups were exposed to 6 MV X-rays at the dose of 15Gy.Rats in Chinese medicine group were treated with ARPD at the dosage of 10 ml·kg-1 ·d-1 once a day,but rats in single radiation group did not receive ARPD treatment.Rats in control group were treated with neither irradiation nor drugs.Five rats of each group were killed and the lung tissues and blood samples were collected at 15,30,60,75,90,105 and 140 d.The pathological changes of lung tissues were observed and the tissue protein and gene expressions of TGF-β1,PAI-1 and collagen type Ⅲ(C Ⅲ) were assayed by Western blot and RT-PCR.ELISA was used to detect serum TGF-β1 and plasma PAI-1.Tissue and serum HYP were determined by acid hydrolysis and alkaline hydrolysis methods respectively.Results Inflammation was found in the lung tissues of all the exposed rats.Obvious pathological lung fibrosis was found at 60 d,the inflammation and the fibrosis in treated group were slighter than those in single radiation group.In Chinese medicine group,the protein and gene expression levels of TGF-β1,PAI-1,C Ⅲ 30 d(Protein:t =2.49-3.74,t =2.63-4.57 and t =2.76-3.83;Gene:t =2.59-4.33,t =2.83-4.62 and t =2.83-3.96,P＜0.05),serum TGF-β1 and plasma PAI-1 15 dlater (t =2.85-6.27 and t =3.69-5.27,P＜0.05),and the levels of tissue and serum HYP60 dlater (t=3.65-4.40 and t =6.56-3.75,P＜0.05),all of them were lower than those in single radiation groups.There were significant positive correlations between tissue TGF-β1 and PAI-1 as well as C Ⅲ (Protein expression:r =0.604,0.759,P ＜0.05;Gene expression:r=0.519,0.816,P＜0.05).Conclusions ARPD may inhibit the pulmonary fibrosis by decreasing the levels of TGF-β1,PAI-1 and C Ⅲ.

Objective To study the calcium metabolism in tacrolimus(FK506)induced rats nephrotoxicity and the preventive effect of calcium channel blocker.Methods Twenty-four Spragueinduced or FK506-induced nephropathy model.Blood creatinine,blood electrolytes,renal tissue histopathology(HE stain)and the change of ultrastructural organization in renal cells by transmission electron microscope were observed.Results The blood creatinine levels of both CsA and FK506 groups [(36.00±2.61)and(34.17±4.54)μmol/L] were significantly higher than those of the FK506+Dilgroup and control group(all P＜0.05).The blood calcium levels of both CsA and FK506 groups (2.00±0.04 and 2.05±0.04 mmol/L) were significantly lower than those of the FK506+Dil group and control group(all P＜0.05).The blood creatinine and calcium levels of FK506+Dil group were not significantly different with those of control group(P＞O.05).Histopathology examination showed cloudy swelling and vacuolization of the renal tubular epithelial cells and intra-cellular mitochondria swelling and vacuolization in the CsA and FK506 groups.However,the pathological changes of the FK506+Dil group were remarkably milder in comparison with the CsA and FK506 groups.Concluum channel blocker,Dil,could prevent the FK506-induced nephrotoxicity.

Objective To discuss the clinical effects of laparoscopic therapy and duodenoscopic therapy in the treatment of biliary severe acute pancreatitis.Methods A total of 140 patients with biliary severe acute pancreatitis(BSAP) in our hospital was assigned into laparoscopic therapy group(60 cases) and duodenoscopic therapy group(80 cases) according to the treatment plan,and indicators in both groups were compared,including acute physiology and chronic health evaluation(APACHE Ⅱ score),WBC,CRP,the operation time,intraoperative blood loss,hospitalization time,hospitalization fee,complication rate,cure rate,mortality rate,abdominal pain disappearing time,body temperature recovered to normal time,blood amylase recovered to normal time,amino acid transaminase(ALT) recovered to normal time,total bilirubin(TBIL) recovered to normal time,alkaline phosphatase(ALP) recovered to normal time.Results APACHE Ⅱ scores and the levels of WBC,TBIL,ALT and CRP in both groups were significantly decreased after surgery(P＜0.05),but there were no statistical differences between the two groups at the same time point after treatment(P＞0.05).The operative time and the intraoperative blood loss in the duodenoscopic therapy group were significantly less than those in laparoscopic therapy group(P＜0.05).There were no statistical differences between the two groups of hospitalization time,complication rate,cure rate,mortality rate,abdominal pain disappearing time,temperature returned to normal time,blood amylase recovered to normal time,ALT recovered to normal time,TBIL recovered to normal time,ALP recovered to normal time(P＞0.05).Conclusion In early stage,both laparoscopic therapy and duodenoscopic therapy can treat BSAP effectively,and their therapeutic effects are nearly similar.

For advanced pulmonary metastases or melanoma, radiotherapy combined with immune checkpoint inhibitor can block the immunosuppression pathway, enhance the antitumor immune response, and significantly improve survival. Stereotactic body radiation therapy ( SBRT ) delivers a large dose of radiation to the tumor target with high precision while sparing irradiation of the surrounding normal tissues. It is suggested that SBRT could be the most appropriate radiotherapy modality to be combined with immunotherapy since it induces the expression of a series of cytokines and immune molecules and is more likely to cause intense immune response and exert an abscopal effect than conventional radiotherapy. Previous studies have explored that total dose and fractionation seem to be important parameters for determining the immune response;the timing of radiation with immunotherapy significantly influences the outcome, and tumor infiltrating lymphocytes, the expression level of programmed death-ligand 1, and mismatch repair defect may be important predicators of the outcome. With appropriate radiotherapy dose and fractionation, the optimal timing of radiation with immunotherapy, and effective predictive markers, a combination of SBRT and immunotherapy may eliminate advanced malignancies while activating the systemic immune response to exert an abscopal effect.

Objective:To assess the bacterial profiles and antimicrobial susceptibility patterns in uropathogens, and help to inform the empiric treatment decisions for urinary tract infection in outpatient settings.Methods:A single institutional retrospective analysis was performed on positive urine cultures from outpatient settings between January 1998 and December 2018. To analyze changes over time, trends analysis were undertaken on bacterial profiles, antimicrobial susceptibility and resistance.Results:A total of 1.172 pathogenic bacteria were isolated after exclusion of duplicate strains originated from the same patient, including 991(84.6%) Gram-negative bacterial strains and 181(15.4%) Gram-positive strains. The most common Gram-negative uropathogens were Escherichia coli (60.8%) and Klebsiella pneumonia (8.1%). Enterococcus faecalis (4.6%) was the predominant Gram-positive strain. The detection rate of Escherichia coli increased significantly, from 50.8% to 63.2% ( χ2=7.978, P=0.046), and no significant difference was observed in the distribution of major uropathogenic bacteria over the 20 years (all P>0.05). The proportion of extended-spectrum β-lactamase (ESBLs) producing strains increased significantly across the 20 years ( P<0.05). The resistance rates of Escherichia coli to amoxicillin and clavulanate potassium, aztreonam, ceftazidime, ciprofloxacin and sulbactam + cefoperazone increased significantly (all P<0.05). All the isolates sustained high susceptibility to tazobactam + piperacillin, amikacin, imipenem and nitrofurantoin (95.0%, 95.7%, 97.9% and 91.1%). Similar to those of Escherichia coli, Klebsiella pneumoniae remained a high and stable sensitivity to tazobactam+piperacillin, amikacin and imipenem during the 20 years (79.1%, 88.0% and 80.3%). However, the proportion of ESBLs producing strains increased significantly ( P<0.05). Among Gram-positive bacteria isolates, the sensitivity rates of Enterococcus faecalis to ampicillin, nitrofurantoin and penicillin G were 100.0%. No vancomycin resistant strain was detected in Gram-positive bacteria. Conclusions:From 1998 to 2018, Escherichia coli and Klebsiella pneumoniae are the most common Gram-negative bacteria uropathogens obtained in outpatient settings. Significant increases of resistance to some antimicrobial agents such as second- and third-generation cephalosporins and fluoroquinolones are observed during the 20 years and high susceptibilities to tazobactam+piperacillin, amikacin, imipenem and nitrofurantoin sustain over time. Local treatment strategies of urinary tract infections on outpatient basis should be made according to epidemiology of drug resistance and individual characteristics to control the spread and curb the prevalence of drug resistant.