ObjectiveTo provide a reference for the establishment and appliance of enzyme reference measurement of CK in China by comparing and analyzing the RELA results of CK in IFCC from 2006-2008. Methods The RELA samples of CK were measured according to the reference procedure for the measurement of catalytic activity concentration of CK (37 ℃) ,which had been published by IFCC. The EP5A2 protocol was used for evaluation of the imprecision and ERM was used for verification of the trueness. Results In RELA 2006, the result of sample A was (9. 896 ±0. 112) μkat/L, and the result of sample B was (4.953 ±0. 120) μkat/L. In RELA 2007, the result of sample A was (2.684 ±0.054)μkat/L, and the result of sample B was (8.798 μ0. 101) μkat/L. In RELA 2008, the result of sample A was (10. 523 ±0. 149) μkat/L,and the result of sample B was (10. 551 ±0. 141) μkat/L. The precision of the CK reference method in the year 2006 to 2008 was 0. 92%, 0. 86% and 0. 88% respectively, each of them is less than 1% and the results of ERMs were consistent with the certified value(1. 68 ± 0. 07)μkat/L,which verify the imprecision and accuracy of the reference method. Conclusions All of the results in the continuous three years were in the range of equivalence limits suggested by IFCC. The CK reference method suggested by IFCC has been established and it is getting better.

Objective In order to study the important role of two specific VEGF receptors(Flt 1 and KDR) in angiogenesis, we design the experiment to detect the gene expression of Flt 1 and KDR in human umbilical vein endothelial cells cultured in hypoxium. Methods We cultured the human umbilical vein endothelial cells for different time and treated it with vascular activate factor, such as endothelin (ET), Nitric oxide (NO), LNNA (inhibitor of NO). The expressive level was analyzed by Northern blot or RT PCR. Results Hypoxia increases flt 1 mRNA in endothelial cells at 6 hours. Administration of ET stimulated an increase in flt 1 mRNA. NO inhibited expression of flt 1 mRNA. LNNA increased its expression. Hypoxia partly affected the expression of KDR. LNNA enhanced the VEGF transcripts. Conclusion Hypoxia up regulated flt 1 gene expression, partly KDR. ET enhanced and NO inhibited flt 1 gene expression.

Objective To investigate the regulation and expression of VEGF gene in human umbilical vein endothelial cells under hypoxia and the influence of vascular activators,such as endothelin(ET),nitric oxide(NO)and NO synthesis inhibitor N-nitro-L-Arginie(L-NNA),on the expression of VEGF.Methods Culture the human umbilical vein endothelial cells and then treat by hypoxia and vascular activators.The expression on VEGF mRNA level and protein level were assayed by northern blots,ELISA and computer figure analysis system respectively.Results When human umbilical vein endothelial cells were subjected to hypoxia for 6 hours,the expression of VEGF began to increase.ET could increase the expression and NO inhibited it,and LNNA affected the expression of the VEGF.VEGF protein concentration in the cell culture media matches the mRNA result.They are 6 hours hypoxia (8^2?1.1)?g/L,ET+6 hours hypoxia(9^37?1^02)?g/L,NO+6 hours hypoxia(2^86?0^91)?g/L,LNNA+6 hours hypoxia(14^75?1^87)?g/L respectively.Conclusion (1)Hypoxia can induce the expression of VEGF in human umbilical vein endothelial cells.(2)ET can increase the inducible effect of hypoxia,NO can inhibit it and LNNA stimulated the expression of VEGF in human umbilical vein endothelial cells.

Pneumonia is one of the most common medical complications after stroke.Stroke-associated pneumonia (SAP) can not only increase the length of hospital stay and medical cost of patients,but also an important risk factor for mortality and morbidity in patients with stroke.All these indicate that the importance of prediction and prevention of SAP.This article reviews the advances in research on the prediction and prevention of SAP.

Objective:This study investigated the clinical characteristics of multiple myeloma with early death in the era of novel drugs. Methods:Medical records from 188 patients diagnosed from January 2009 to December 2015 were retrospectively reviewed, showing that early death occurred in 19 patients. Early death was defined as death by any cause within the first year after diagnosis. Results:(1) Early mortality was 10.1%, and the median age was 67 years old (range:40-84 years). Eight cases presented IgG type, and 11 cases were non-IgG type. All 19 patients were diagnosed to be at stageⅢin accordance with the Durie–Salmon staging system, and renal insufficiency occurred in 10 patients. In accordance with the International Staging System (ISS), four patients were diagnosed to be at stageⅡ, whereas 15 other patients were at stageⅢ. Extramedullary plasmacytoma (EMP) occurred in six cases, whereas 10 cases pre-sented high-risk patients with cytogenetic abnormalities. Elevated lactate dehydrogenase (LDH) was found in five cases, amyloidosis was detected in three patients, and secondary plasma cell leukemia was observed in two cases. The median score of performance sta-tus (KPS) was 70 (range: 20-80). A total of 16 patients were treated with bortezomib, and 3 patients were treated with CADT. (2) Among the 13 patients who were evaluated, the overall response rate was 46.2%(6/13), and the complete response (CR) and near-CR rate was 7.7%(1/13). (3) The median overall survival was 3 (1-11.5) months, although the two patients with secondary plasma cell leu-kemia survived for less than 2 months. (4) Eight patients died of disease progression (42.1%), eight patients died of severe infections (42.1%), and three patients died of thrombotic events. Conclusion:The important causes of early death include the following:high-risk cytogenetics, elevated LDH, EMP, amyloidosis, advanced age, poor performance status, and serious complications during treat-ment. In the era of novel drugs, we should improve early diagnosis rates and explore individualized treatment for high-risk multiple my-eloma for the benefit of a wide range of patients.

Objective To investigate the efficacy and outcome in newly diagnosed multiple myeloma (MM) patients with renal insufficiency using bortezomib-or thalidomide-based regimens as front line treatment.Method Sixty-nine newly diagnosed MM patients with renal insufficiency were retrospectively analyzed from August 2006 to August 2014.Results ① Among thirty-nine patients with bortezomib based regimens (the bortezomib group),the overall response rate (ORR) was 89.7％ and complete response (CR) plus near CR(nCR) rate was 41.0％.By contrast,among thirty patients with thalidomide based regimens (the thalidomide group),the ORR was 83.3％ and CR + nCR rate was 26.7％.There was no significant difference of either ORR or CR + nCR rate between bortezomib and thalidomide groups.② The improvement rate of renal function in bortezomib group and thalidomide group were 87.2％ and 60.0％respectively (P =0.012).The median duration time of renal injury was 45 days in 52 patients with renal function improved,which was significantly shorter compared with 222 days in 17 patients without improvement (P ＜ 0.05).There was no difference of median serum creatinine and creatinine clearance rate between the two groups.③ The median progression-free survival (PFS) and the overall survival (OS) were 18 and 33.5 months,respectively in all patients.The three-year and five-year OS rates were 57％ and 17％,respectively.The median PFS was 19 months in bortezomib group,while it was only 12 months in thalidomide group (P =0.023).The median OS were 36.5 months and 25.5 months respectively,which was no difference (P =0.285).Conclusions The newly diagnosed MM patients with renal insufficiency could get higher ORR and the longer PFS using bortezomib-containing regimens as initial therapy.Meanwhile the improvement rate of renal function and the living quality in patients with bortezomib are better compared with those with thalidomide based treatment.

We have found the parallel relationship between the live cell number of several tumor cell lines and the formation of MTT formazan with MTT colorimetric assay. The MTT colorimetric assay was compared with ~3H-Tdr incorporation assay for the proliferation of mouse spleen cells induced by mitogens or the activity of rat/mouse IL2,the results suggested that the ~3H-Tdr incorporation assay can be replaced by MTT colorimetric assay. MTT colorimetric assay have the advantages of simplicity, rapidity,save any radioisotope and no specific equivepment, etc, so under some conditions it would be a useful method for measuring cell proliferation or cytotoxicity in a laboratory.

Objective To investigate the expression of resistin in rats of acute pancreatitis and the relationship between its expression and pancreatic pathologic changes. Methods Forty Sprague-Dawlev rats were randomly divided into four groups:control group,sham operation group,AEP group and ANP group. AEP model was induced by intraperitoneal injection of cerulein and ANP model was induced bv intrapefitoneal injection of L-arginine while the control group received no treatment,and the sham operation grouD rleceived same volume injections of 0.9%saline solution.The serum amylase,C-reactive protein,TNF-alpha.IL-1 beta were determined by using ELISA,pancreas/body weight(g/kg)ratio was recorded,pancreatic pathology was examinated,the expression of resistin mRNA and protein in pancreas was detected by real-time PCR and immunohistochemistry. Results The levels of amylase,pancreas/body weight ratio.pancreatic pathology score,expression of resistin mRNA,IL-1-beta,TNF-alpha and C-reactive protein in AEP group were(4377±343)U/L,8.67 ±1.43,5.39 ±0.26,2.04 ±0.19,(10.21 ±1.34)ng/ml,(184.18±45.24)pg/ml, (194.24 ±44.81)pg/ml,(3586±63)ng/ml,respectively;and the corresponding value8 in ANP group were (6750±322)U/L.9.33±1.76,7.81±0.28,3.29±0.30,(15.14±0.84)ng/ml,(349.31±94.54)pg/ml,(315.59±37.04)pg/ml,(4345±244)ng/ml,respectively;which were significantly higher than(1442±183)U/L,4.34±0.42,1.10±0.21,0.88±0.08,(5.13±0.74)ng/ml,(108.74±31.03)pg/ml,(106.44±21.31)pg/ml,(2895±165)ng/ml in sham operation group(P<0.01 or P<0.05).Resistin wag positively correlated with the increase of C-reactive protein,TNF-alpha,IL-1 beta and the pathologic score of pancreas,respectively(r=0.711,0.812,0.794,0.812,P<0.01).Conclusions Resistin was correlated with the pathogenesis and development of acute pancreatitis,and may be useful to predict the severity of acute pancreatitis.

Objective To investigate the value of 1q21 amplification in newly diagnosed myeloma patients.Methods Fifty-two cases of newly diagnosed multiple myeloma from June 2008 to June 2010 were enrolled.Fluorescence in situ hybridization (FISH) was used to detect the 1q21 amplification,and the clinical characteristics and treatment response were analyzed.Results 1q21 amplification was discovered in 30 of 52 patients (57.7 ％),Clinical characteristics such as gender,malignant pleural effusion,extramedullary plasmacytoma,bone destruction,β2 microglobulin,ALB,hemoglobin,blood calcium,plasma cell proportion,clinical stage seemed to have no correlation with 1q21 amplification.The 52 patients all received bortezomibbased regimens.The response rates were not significant difference between patients with and without 1q21 amplification,the OS was also not significant difference [26 months (6-30 months) vs 30 months (12-85 months),P =0.409],but the patients with presence of 1q21 gain resulted in significantly shorter PFS [8 months (1-30 months) vs 20 months (3--48 months),P=0.019].Multivariate analysis showed 1q21 with more than two additional genetic abnormalities was an independent prognostic predictor (P =0.031).Conclusion 1q21 amplification is one of the adverse prognostic predictors,the response rate is not significant difference between patients with and without 1q21 amplification in bortezomib-based group,but the 1q21 amplification could result in significantly shortened PFS.

Objective To present our experience of dealing with complete penile amputation.Methods Two cases of penile complete amputation were reported.The first case was a 34-year-old man,suffered amputation of the penis approximately 2.5 cm distal from the pubic area with a sharp knife.3.5hours later,the patient was transferred to our hospital.The urethra mucosa and corpus spongiosum were anastomosed.The cavernous body of the penis was reattached by suturing the tunics albuginea of each corpus cavernosum to the corresponding proximal segment.One dorssl artery,two dorsal veins,and dorsal nerve were anastomosed under a 10 × microscope with interrupted 9-0 nylon nonabsorbable sutures.The second case was a 25-year-old man,presented to the emergency room 15 hours after distal penile amputation,which had 2 wounds as a result of self-mutilation caused by psychiatric problems.The urethra mucosa and corpus spongiosum were anastomosed.The cavernous body of the penis was reattached by suturing the tunics albuginea of each corpus cavemosum to the corresponding proximal segment using 4-0 polyglactic acid sutures.Results In the first case,the tourniquet was released after replantation,and the distal penis appeared to revascularize,as noted by the gradual increase in redness and size.An arterial pulse was detected,and the superficial penile veins displayed normal turgor,and no bleeding was found.On postoperative day 3,the penile skin started to necrotize.On day 12,the necrotic skin was superficially debrided,and a fistula was observed in the corresponding urethral segment.Two weeks later,the fistula was sutured with 4-0 interrupted synthetic absorbable suture,and a transposition flap to embed the whole injured penis shaft was created from the proximal scrotal skin.The glans was exposed.Two months after the second operation,the embedded penis was released from the scrotum.After follow-up of two years,the patient had glans re-epithelialization with normal voiding,sensation,and erections.In the second case,the glans was still pink,but the penile skin started to necrotize on postoperative day 3.On day 14,serious infections were noted,the necrotic skin was superficially debrided,and the amputated penis was relieved.Conclusions Prompt diagnosis and early treatment are essential to avoid the potential complications of ischemic necrosis and autoamputation.Venous outflow is a critical factor for success of replantation.Microsurgical reanastomosing of the dorsal penile vein,penile arteries,and dorsal nerves can be identified as the standard method for penile replantation.The bipedicled scrotal flap can provide adequate skin cover for penis defects.