A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for quantification of sitagliptin in human plasma and urine had been developed. This method was applied to the pharmacokinetics study of sitagliptin tablet after single- and multiple-dosing in Chinese population. Plasma samples were prepared by a liquid-liquid extracted method, and urine samples were diluted. Compounds were analyzed by multiple reaction monitoring (MRM) mode with a electrospray ionization (ESI) interface. Mobile phase consisted of methanol and water (85 : 15, v/v). The linear concentration range of calibration curve was 0.5-1 000 ng.mL-1. and 0.2-100 µg.mL , intra-run/between-run accuracy was 98.98%-103.69% and 97.63%-102.29%, intra-run/between-run precision was <5.51% and 4.26% for plasma and urine sample, respectively. The stability of sitagliptin stock solution was tested for 55 days at -30 °C. Sitagliptin was stable when stored under the following conditions: 24 hours in the autosampler after sample preparation; 24 hours at room temperature, after 3 freeze and thaw cycles (from -30 °C to room temperature), 40 days at -30 °C for plasma and urine samples. The absolute recovery in plasma was 71.1%, and no matrix effect was founded. This method was proved simple, specific, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.
Calibration ; Chromatography, Liquid ; Humans ; Liquid-Liquid Extraction ; Sitagliptin Phosphate ; blood ; pharmacokinetics ; urine ; Tandem Mass Spectrometry

OBJECTIVE:To improve the management on drug supply in our hospital by using the Six sigma method. METHODS: The quality control of the management on drug supply is achieved by practicing the five steps of Six sigma: concept, measurement, analysis, improvement and control. RESULTS & CONCLUSIONS: Six sigma method did effectively bring down the defect rate of drug supply from 6.66% before the introduction of the method to 0.59% after and raise the degree of satisfaction to the work of medicine provision from 80% before the introduction of the method to 95% after.

Aim Toinvestigatetheroleofbrain-de-rived neurotrophic factor(BDNF)-tyrosine receptor ki-nase B (trkB ) signaling pathway in the therapeutic effects of ketamine on diabetic neuropathic pain.Meth-ods Forty-eightWistarrats,aged3months,weighing 200~250 g,were equally randomized into 4 groups(n=12 ):control group (C group ), saline group (S group),ketamine group (K group)and ketamine +ANA-12 group (KA group ).Rats in S,K and KA groups were intraperitoneally injected with a single of streptozotocin(STZ)65 mg·kg-1 to construct diabetic neuropathic pain model.After twenty-eight days,rats in S,K and KA groups were intraperitoneally injected with saline, ketamine 10 mg·kg-1 and ketamine 10 mg·kg-1 +ANA-12 0. 5 mg·kg-1 for consecutive 7 days, respectively. On the 8th day, mechanical withdrawal threshold(MWT)of rats was measured.Af-ter that,the rats were immediately sacrificed,and dor-sal ganglion of lumbar spine and prefrontal cortex (PFC)were harvested for measuring BDNF,p-trkB/trkB,synaptophysin and spine density by Western blot andglogistaining.Results ComparedwithCgroup, rats in S group significantly decreased MWT,BDNF, p-trkB/trkB,synaptophysin and spine density in dorsal ganglion and PFC (P <0. 05 ).Compared with S group,rats in K group showed a significant increase of MWT,BDNF,p-trkB/trkB,synaptophysin and spine density in the all observed regions(P<0. 05 ).On the contrary,rats in KA group showed a significant de-crease of MWT and BDNF,p-trkB/trkB,synaptophys-in and spine density as compared with K group in all regions(P<0. 05 ).Furthermore,BDNF was positive-ly correlated with spine density in all regions (P <0.05).Conclusion BDNF-trkBsignalingpathway mediates ketamine-induced therapeutic effects in dia-betic neuropathic pain.

Objective To observe the specific changes of content of cyclic guanosine monophosphate (cGMP) around meridian skin and specific influences of electroacupunture Neiguan (PC6) on cGMP of rats with acute myocardial ischemia (AMI). Methods With 6 male Wistar rats as normal group, 18 rats were made AMI model and divided into model group, high-frequency group and low-frequency group (n=6). The high-frequency and low-frequency group were given electroacupuncture in bilateral PC6 with 100 and 2 Hz for 20 min respectively. The content of cGMP in skin around PC6, Ximen (PC4), Tianquan (PC2), Waiguan (TE5) and Zusanli (ST36) was detected by using enzyme-linked immunosorbent method after treatment. Results In model group, cGMP content increased significantly around PC6, PC2, TE5 and ST36 compared with normal group (P <0.05,P <0.001). After the treatment with high-frequency or low-frequency electroacupunture in PC6, cGMP content decreased significantly in skin around PC6, PC2, TE5 and ST36 compared with model group (P <0.05, P <0.001). Conclusion High-frequency electroacupunture or high-frequency electroacupunture used in PC6 can protect rats from the injury of AMI. The mechanism may be related to the decrease of cGMP content around the acupoint areas in heart-related meridian.

Adolescent ; Cholesteatoma ; microbiology ; Ear, External ; Fistula ; etiology ; Humans ; Male ; Parotid Gland

OBJECTIVETo investigate the relationship between (GT)n polymorphism and esophageal cancer by analyzing the connection between microsatellite polymorphisms in the promoter of heme oxygenase-1 and the clinicopathological characteristics of esophageal squamous cell carcinoma (ESCC) in Han chinese population.

METHODSThe (GT)n repeats in HO-1 gene in 83 male and 43 female hospital-based patients with ESCC (aged between 40 and 79 years with a mean of (61 ± 8) years) and 134 healthy control individuals were obtained by DNA sequencing. Polymorphisms of the (GT)n repeats were generally grouped into three classes based on allele frequencies: class S alleles (<25 repeats), class M alleles (25 to 29 repeats), and class L alleles (≥30 repeats). The correlation between susceptibility and clinicopathological characteristics of ESCC were analyzed by χ2 test. For in vitro experiments, the transient-transfection assay was performed to explore the correlation between different lengths of (GT)n repeats and promoter activity by assessing the promoter activities of HO-1 gene in cultured Ecal09 cells treated with H2O2 by analysis of cariance.

RESULTSHigher frequencies of L-allele (25. 8% vs. 14. 9%, χ2 = 9. 520, P = 0. 002), L-allele carrier (41. 3% vs. 27. 6%, χ2 = 5. 381 , P = 0. 020) were found in patients with ESCC. Furthermore, the lymphatic metastasis rate (63. 5% vs. 41. 8%, χ = 5. 685, P = 0. 017) and the detection rate of poorly differentiated ESCC cell (53. 8% vs. 28. 4%, χ2 = 8. 335, P = 0. 004) was significantly higher in L-allele carriers compared to non-L-allele carriers. In transfection experiments, promoter activities of 5'-flanking regions of the HO-1 gene in Eca109 cells transfected with the recombinant gene carrying (GT)16 repeat after treatment with H2O2 increased (F = 23. 615,P = 0. 008). In H2O treated control group, compared to (GT)26 and (GT)36, the basal promoter activities of HO-1 gene carrying (GT)16 repeat increased (F =41. 376, P = 0. 003; F = 50. 761, P = 0. 002).

CONCLUSIONThe long (GT)n repeats of HO-1 gene promoter can increase the susceptibility of esophageal squamous cell carcinoma and the risk of lymphatic metastasis.

Alleles ; Asian Continental Ancestry Group ; Carcinoma, Squamous Cell ; etiology ; pathology ; Esophageal Neoplasms ; etiology ; pathology ; Female ; Gene Frequency ; Heme Oxygenase-1 ; genetics ; Humans ; Hydrogen Peroxide ; Lymphatic Metastasis ; Male ; Microsatellite Repeats ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Risk Factors ; Transfection

Aim To explore the role of PI3 K/Akt/Sirt1 pathway in cardioprotection of hydrogen sulfide ( H2 S ) postconditioning against ischemia/reperfusion ( I/R) injury. Methods Langendorff perfusion appa-ratus was used to build an isolated rat myocardial I/R model. Isolated rat hearts were subjected to 30 min global ischemia followed by 60 min reperfusion after 20 min of equilibrium. 60 male SD rats were randomly di-vided into 5 groups(n=12):control group(Control), ischemia/reperfusion group( I/R) , H2 S postcondition-ing group( H2 S) , inhibitor LY294002 group( LY) and H2 S with inhibitor group( H2 S+LY) . The left ventric-ular diastolic pressure ( LVEDP ) , the left ventricular developed pressure(LVDP), the maximum rate of in-crease or decrease of left ventricular pressure ( ± dp/dtmax ) were registered at the end of 20 min equilibri-um, 30 and 60 min of reperfusion separately. Triphe-nyl tetrazolium chloride( TTC) staining was used to de-termine the myocardial infarct size. The levels of Sirt1 and PGC-1 mRNA were tested using real-time PCR. The expressions of Sirt1 and PGC-1αwere detected with Western blot analysis. Immunohistochemical method was used to determine the location of Sirt1 . Results There were no differences in equilibrium hemodynamics observed between the experimental groups(P>0. 05). At the end of reperfusion, compared with I/R group, H2 S group had obviously ameliorated functional recov-ery and significantly decreased the myocardial infarct size(26. 9 ± 4. 9)% vs(48. 9 ± 5. 6)%(P <0. 05). Meanwhile, the expression of Sirt1 and PGC-1α in-creased significantly. However,LY294002 reversed the cardioprotective effects provided by hydrogen sulfide postconditioning and reduced the level of Sirt1 and PGC-1α, the percentage of Sirt1-positive nuclei. Con-clusion PI3 K/Akt/Sirt1 signaling pathway mediates the hydrogen sulfide postconditioning-induced protec-tion against I/R injury.

Objective To evaluate the feasibility and efficacy of ultrasound-guided popliteal vein catheter thrombolysis for acute deep venous thrombosis of the lower extremity.Methods Clinical data were retrospectively analyzed on 120 patients of unilateral acute lower extremity deep vein thrombosis from April 2010 to April 2013.60 cases were included in systemic thrombolytic group; 60 cases into catheter directed thrombolysis group.Thrombolysis rate and limb swelling reduction rate were calculated and compared between the two groups.Results Due to limb swelling rate,in systemic thrombolysis group:Thigh swelling reduction rate was (77 ± 9) ％ ; Leg swelling reduction rate was (70 ± 11) ％,while in catheter directed thrombolys group,that was (87 ± 5) ％,and (80 ± 9) ％,respectively (P ＜ 0.05).The thrombolysis rate in systemic group was (59 ± 14)％,that was (71 ± 13)％ in catheter directed thrombolysis group (P ＜ 0.05).Conclusions Ultrasound guided precutaneous catheter popliteal vein thrombolysis significantly improves the short-term outcome of deep venous thrombosis in terms of leg swelling reduction rate and thrombolysis reduction rate.

Abstract: Objective　To detect the antibody levels of hantavirus in serum samples from patients suspected with hemorrhagic fever with renal syndrome (HFRS) in Heilongjiang Province from 2019 to 2021, and to provide scientific basis for the prevention and control of disease. Methods　Enzyme-linked immunosorbent assays (ELISA) were used to detect the IgM antibodies to hantavirus in serum samples collected from suspected patients with HFRS in the acute-phase, and IgM and IgG antibody in convalescent-phase serum samples. The positive rate of IgM antibody in acute-phase serum samples of patients in different years was analyzed with χ2 test by SPSS 19.0, and the data were sorted out and analyzed about patients' gender, occupation, age, date of onset and interval from onset to initial diagnosis by EpiData 3.1, Excel 2003 software. Results　A total of 351 acute-phase serum samples and 208 convalescent-phase serum samples were detected in patients suspected with HFRS, respectively. There were 317 positive IgM antibodies of serum samples in the acute stage, with the positive rate of 90.31%. There was no significant difference in the positive rate of IgM antibodies in the acute stage between different years (χ2=0.895, P=0.639). T The IgM antibodies and IgG antibodies were positive in 32 (15.39%) and 28 (13.46%) of the convalescent-phase serum samples, respectively. Moreover, 148 patients (71.15%) were double-positive for IgM and IgG antibodies at the convalescent stage. The ratio of male to female patients was 4.56∶1, for which male patients were much more than female patients. Occupation was dominated by farmers (253 cases, 79.81%), followed by workers (19 cases, 5.99%) and the unemployed (17 cases, 5.36%), respectively. The age of patients ranged from 10 to 88 years old, with a median age of 49 years old. Most of the patients were in the age group from 30 years old to 60 years old (209 cases, 65.93%), among which the age group from 40 years old to 50 years old (86 cases, 27.13%) had the highest proportion, and the age group from 60 years old to 90 years old had a proportion of 20.18% (19 cases). May and November were the peak periods of HFRS in Heilongjiang Province. The median interval between onset and initial diagnosis was 4 days. Conclusions　There is a gap of about 10% between the clinical diagnosis of HFRS cases and the confirmed cases detected by laboratory in Heilongjiang Province from 2019 to 2021. The virus-specific detection results are important for confirming the diagnosis of local patients with HFRS.

OBJECTIVETo summarize the clinical effect of avulsed skin replantation of hand and foot via vacuum sealing drainage (VSD) combing low temperature technique.

METHODSFrom March 2012 to October 2013,13 cases with avulsed skin replantation of hand foot using combined technique included 8 males and 5 females with an average age of 32 years old ranging from 18 to 62 years. The time from injury to hospital was 1 to 4 hours (2.4 hour in average). The reasons of injury included machine injury in 7 cases and rolling over by cars in 6 cases. The parts of injuried involved finger in 2 cases,back of the hand in 5 cases and dorsum of foot in 6 cases. The area of avulsed skin was 5 cm x 6 cm to 12 cm x 16 cm,tendon and bone exposure was found in 4 cases. VSD was operated in all patients and the avulsed skin was refrigerated in the temperature of -4 °C or -80 °C. After 4 days, the skin stored in the -4 °C was replanted to the wounded place in 5 cases and in 3 cases the skin was planted to the donor site of flap. The skin stored in the -80 °C was replanted in 4 cases after 7 or 8 days, 1 case after 45 days.

RESULTSOf the 13 cases, 1 case of degloved injury from lower leg to dorsal foot,the replanted skin was necrosis completely; 1 case of degloving injury with fourth finger,the skin which replanted after 45 days survived approximately 30%,cured after skin-graft many times. In the other cases, the survival area of replanted skin was more than 85%, all cured after dressing. According to the standard of skin survival area evaluation by Jia et al, 11 cases showed excellent, 1 showed medium and 1 showed inferior. There were no complication about grafted skin rupture after the skin survived in 11 patients,after 4 to 22 months follow-up, the resiliency of grafted skin showed good. Sensation recovery was measured by BMRC standard: 3 cases of S3, 5 cases of S3, 3 cases of S2.

CONCLUSIONVSD combining lower temperature technique in skin replantation provides time and space for wound preparation and treatment plan for the patients who need second surgery, especially for the large area skin degloving,this method could utilize the degloved skin efficiently, decrease the donor site area, alleviate the pain and financial burden,reduce the scar formation of donor site and impediment.

Adolescent ; Adult ; Cryopreservation ; methods ; Drainage ; instrumentation ; methods ; Female ; Foot Injuries ; surgery ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Replantation ; Skin ; injuries ; Skin Transplantation ; Young Adult