Objective To analyze reason and countermeasures of unqualified blood specimens ,improve the qualified rate of sam-ples ,to strengthen the quality control before analysis .Methods A retrospective statistical analysis were conducted to analyze the characteristics of the unqualified specimen and reasons from January 2013 to June 2014 .Results A total of 11 024 unqualified spec-imens accepted in the inspection center from January 2013 to June 2014 ,accounting for 0 .331% of reasons of unqualified specimens including hemolysis(26 .7% ) ,blood coagulation(25 .8% ) .The unqualified specimen in surgical system was higher than that in med-ical system .Conclusion Control specimen qualified rate system should be established in clinical laboratory ,the fraction defective samples should be reduced through continuous analysis and communication with clinical medical personnel to ensure the quality be-fore analyzing .

Objective To explore the clinical features and CT characteristics of invasive pulmonary aspergillosis(IPA) in children with acute leukemia.Methods CT scanning of chest was performed in 13 cases with IPA proved clinically and sputum culture in children with acute leukemia.All cases followed up by CT 4 to 10 times.The clinical features and CT findings were analyzed retrospectively.Results The early CT findings included single nodule in 2 cases,multi-nodular shadow in 10 case and sub-pleural wedge-shaped consolidation in 5 cases,of these 5 cases,4 accompanied multi-nodules.Halo sign around the nodules in 9 cases.The follow-up CT showed air crescent sign or cavity in 11 cases,halo sign around the nodules in 7 cases and around cavity in 2 cases.Conclusion IPA has some characteristic manifestations on CT.CT is of important value in observing the evolution and evaluating the prognosis for the patients with IPA.

Objective To investigate the expression of miRNAs in endoscopic ultrasound (EUS)-guided fine needle pancreatic aspirates (FNA) and its clinical significance. Methods The expression of miRNA-210, miRNA-21, miRNA-196a, miRNA-181a, miRNA-181b, miRNA-155 and miRNA-16 in EUS-FNA specimen of 23 pancreatic cancer and 13 benign pancreatic masses was detected with quantitative real-time polymerase chain reaction (qRT-PCR) assays and its clinical significance was analyzed. Results The relatively expression quantity of miRNA-210, miRNA-21,miRNA-196a, miRNA-181a and miRNA-181b (0.86±1.10, 0.69±0.64, 0.32±2.50, 0.16±0.83and 0.56 ±0.88, respectively) was significant higher in pancreatic cancer than those in the benign pancreatic masses (-0.11±0.98,-0.03±0.97,-1.50±1.40,-0.53±1.10 and -0.28±1.10,respectively) (P value was 0. 012, 0. 011, 0. 024, 0. 036 and 0.015, respectively). The relatively expression quantity of miRNA-16 and miRNA-155 in pancreatic cancer and benign pancreatic masses was - 0.11 ± 0.69, 0.08 ± 1.04, - 0.73 ± 1.26 and - 0.19 ± 1.19 respectively, no statistic difference (P value was 0. 067 and 0. 467). The high expression of miRNA196a was positively correlated with pancreatic cancer lymphatic metastasis and clinical TNM staging. Conclusion There were multi miRNAs with abnormal high expression in the pancreatic cancer. Of those, miRNA-196a was associated with the clinical malignant features of pancreatic cancer.

Aim To observe the analgesic effect of oxymatrine(OMT)and its mechanism.Methods A peripheral mononeuropathy was produced in adult mice by placing loosely constrictive ligatures around the common sciatic nerve.The antinociceptive effects of the OMT were assessed in mechanical allodynia and cold allodynia tests.The CAMKII inhibitor KN-93 and AIP were adopted to investigate the influence of OMT on the analgesic effect and analyze its analgesic mecha-nisms.Western blot was used to evaluate the expres-sions of tCaMKII and pCaMKII protein.Results The intraperitoneal administration of OMT (1 60,80 mg· kg -1 )increased the paw withdrawal threshold in the
mechanical allodynia test (P <0.05 ),OMT (1 60, 80,40 mg·kg -1 ,ip)remarkably decreased the paw lifts in the cold allodynia test (P <0.05).Ith KN-93 (1 .25 μg/site),AIP (0.02 μg/site)significantly en-hanced the analgesic effect of OMT (35 mg·kg -1 ) (P <0.01 ).Protein expression of pCaMKII was de-creased by OMT(1 60 mg·kg -1 ).Conclusion OMT has significant protective effects on chronic constriction injury(CCI)in mice,and the effective mechanism of OMT inhibits the expression of CaMKII receptor.

Objective To explore the characteristic of sleep disorders in the initial stage of withdrawal and their relationships with alcohol craving levels in alcohol dependence (AD) patients,and provide support for diagnosis and prevention of re-drinking.Methods Thirty-two AD inpatients were assigned to AD group and 20 male healthy volunteers to control group.Alcohol craving was assessed with the Pennsylvania Alcohol Craving Scale (PACS) within the 2nd week after alcohol withdrawal for AD patients,and then the whole-night polysomnogram (PSG) tracings were conducted on the day of the night.Results The five item scores of the PACS were from 3.48 to 4.26 in AD patients.The sleep latency was(42.48±22.42) min,total sleep time was(289.61± 103.22)min,sleep efficiency was(71.45± 19.86) ％,S1 sleep was (23.47± 11.07) ％,arousal frequencies was (8.01 ± 2.77),S3+4 sleep was(6.26±5.35)％ in AD group.Compared with control group((19.65±8.57) min,(407.33±21.29) min,(81.52 ± 6.46) ％,(8.79± 1.83) ％,(2.17 ± 1.04),(15.87 ± 5.24) ％ respectively),the differences had statistical significances(t=2.206-9.082,P＜ 0.05-0.001).Alcohol craving levels were positively related to sleep latency,arousal frequencies and S1 sleep (r=0.424-0.898,P＜0.05-0.01) and negatively to total sleep time,sleep efficiency and S3+4 sleep (r=-0.416--0.662,P＜0.05-0.01) in AD group.Conclusion AD patients have sleep continuity and structure disturbances in the initial stage of alcohol withdrawal,sleep continuity and structure disturbances are related to alcohol craving.Improvements of sleep disorders should be paid during clinical alcohol dependence treatment.

Objective To evaluate the correlation between homologous recombination repair protein RAD51 and methotrexate-enhanced radiosensitivity.Methods Western blot and RT-PCR assays were used to detect RAD51 expression in HOS osteosarcoma cells exposed to γ-ray irradiation alone and in combination with methotrexate.Colony formation assay was used to test the survival fraction of HOS cells exposed to γ-rays and methotrexate.Results Methotrexate inhibited both protein and RNA expressions of RAD51,and the combination of radiation and methotrexate enhanced the inhibition of RAD51 expression.Moreover,transfection of cells with RAD51 gene decreased cellular sensitivity to methotrexate and γ-rays.The sensitizer enhancerment ratios after irradiation in combination with methotrexate were 1.51 and 0.99,respectively.Methotrenate was a preferred radiosensitizer to HOS cell.Conclusions RAD51 might be involved in the methotrexate-enhanced radiosensitivity.

Objective To investigate the effects of microRNA-196a(miR-196a) inhibitory sequences transfection on HOXB8 expression in PANC1 cells.Methods PANC1 cells were divided into control group,miR-196a inhibitory sequences group and siRNA control group.Liposomal transfection method was applied to transfect miR 196a inhibitory sequences and siRNA control into PANC1 cells.RT-PCR and Western blot were used to detect the expressions of miR-196a and HOXB8 mRNA and protein.Results After miR-196a inhibitory sequences transfection,when compared with that of siRNA control group,the expression of miR-196a was significantly decreased (0.05 ± 0.054 vs.0.839 ± 0.025,t =3.12,P ＜0.05) ; and the expression of HOXB8 mRNA was significantly increased by 1.57 folds (2.20 ± 0.07 vs.1.29 ± 0.10,t =3.86,P ＜ 0.05),the expression of HOXB8 protein was also obviously increased (0.90 ± 0.03 vs.0.40 ± 0.10,t =3.11,P ＜ 0.05).Conclusions MicroRNA-196a down-regulates the expression of HOXB8.

Objective To investigate the clinical effects of pars repair by the pedicle screws,laminal hooks and auto iliac bone graft through the Wiltse approach via Quadrant channel for the treatment of the adolescent spodylolysis.Methods From November 2007 to April 2011,pars defects of 11 patients (10 males and 1 female) were repaired with pedicle screws,laminal hooks and auto iliac bone graft via Quadrant channel through the Wiltse approach.The mean age of 11 patients was 17.3 years (range 15-19).Mean course was 9.9 months (range 6-14 months).All patients have bilateral pars defect of L5.There was no poroma across the pars that have been proved by X-ray film and 3-D CT before the operation.There were 4 patients with Ⅰ grade spondylolisthesis.The degeneration of the intervertebral disc assessed by Pfirrmann grade of all patients was no more than Ⅰ grade.Visual analogue scale (VAS) and Oswestry disability index (ODI) were compared before and after operation.The fusion of pars defect was observed by X-ray film and 3-D CT.Results The mean operation time was 85.5±12.3 min (range 70-110 min); intraoperatve blood loss was 80-150 ml (mean 105 ± 19.4 ml).Two patients were lost in follow-up after operation and the others had been followed up for 12 to 24 months (mean 16 months).All patients had significant pain relief and function recovery after operation.Mean postoperative VAS score was 1.5± 1.0 compared with 4.5± 1.0 of preoperation (t=4.603,P=0.002).ODI was decreased from 49.3％± 14.2％ of preoperation to 12.0％±4.9％ of the last follow-up (t=7.344,P=0.000).With the Macnab grades,7 patients are excellent,2 patients are good.3-D CT showed that all pars defect were fused in the last follow-up.Conclusion Pars repair by the pedicle screws,laminal hooks and auto iliac bone graft through the Wiltse approach via Quadrant channel can protect the paravertebral muscle,relief patients from pain,and improve function.

Objective To analyse clinical application of hepatitis B virus core antibody(HBcAb)detected by using the chemilu-minescence microparticle immunoassay.Methods A total of 1 6 830 specimen with positive HBcAb detected by using the two pairs of semi-hepatitis test from January 2012 to November 2014 were collected,and divided into three groups according to the cut off in-dex(COI)of detection results of HBcAb,including group 1.0-<9.0,group 9.0-<1 1.0 and group COI≥1 1.0,and detection re-sults were statistically analysed.The hepatitis B virus(HBV)DNA test was carried out in specimen with negative hepatitis B surface antigen(HBsAg)and hepatitis B surface antibody (HBsAb)and COI≥1 1.0.Results The detection rate of HBsAg(+)HBsAb(-) (13.84%)was significantly higher than other expression patterns in group ≥1 1.0(P <0.05).There was no statistically significant differences in positive rate among all expression patterns of HBsAg and HBsAb in the group 9.0-<1 1.0(P >0.05).The detec-tion rate of HBsAg(+)HBsAb(-)of group 9.0-<1 1.0 was significantly lower than that of the other two groups(P <0.05).A total of 304 specimen were HBsAg(-)HBsAb(-)and COI≥1 1,among them 64 specimen were HBV DNA postive and the posi-tive rate was 21.0%.Conclusion In the detection of HBcAb,COI≥1 1 and 1.0-<9.0 could be reference indicators for diagnosiing current and past HBV infection respectively,which should be combined with other laboratory indicators of HBV clinical data for comprehensive analysis.

The cyclotides are a family of cyclic "mini" proteins that occur in Violaceae, Rubiaceae and Cucurbitaceae plant families and contain a head-to-tail cyclic backbone and a cystine knot arranged by three disulfide bonds. To study the natural cyclotides of V tianshanica, dried herb was extracted with 50% ethanol, and the concentrated aqueous extract was subjected to a solvent-solvent partitioning between water and hexane, ethyl acetate and n-butanol, separately. The n-butanol extract containing cyclotides was subjected to column chromatography over Sephadex LH-20, eluted with 30% methanol. The subfractions were directly reduced by DTT and analyzed by reverse-phase HPLC. The peaks with different retention times were shown on the profile of RP-HPLC and collected. The cyclotides were speculated based on masses range from 3 000 to 3 500 Da. The purified cyclotides were reduced with DTT, alkylated with iodoacetamide, and then were cleaved with endoproteinase Glu-C, endoproteinase Lys-C and Trypsin, separately. The digested peptides were purified on RP-HPLC and analyzed on MALDI TOF/TOF analyzer. A new cyclotide, cycloviolacin T1 and a reported cyclotide varv E were systemically determined using MALDI TOF/TOF system. So the method for the isolation and characterization of cyclotides was quickly built up in succession.