Objective To develop an improved enzymatic method for assaying creatinine in serum and urine. Methods Cratinine iminhydrolase catalyzes the conversion of creatinine to N-methylhydantoin and ammonia,the later combines with 2-oxoglutarate and NADPH in the presence of glutamate dehydrogenase.Before beginning the reaction sequence,endogenous ammonia is removed by an “ammonia scavenger system”that involves the same auxiliary reaction.The NADPH and 2-Oxoglutarate consumed are restored through a reaction catalyzed by isocitrate dehydrogenase.The activity of this strictly magnesium-dependent enzyme is completely blocked by magnesium-complexing reagent that acts promptly whenever the reaction has started with creatinine iminohydrolase as startor.The reaction is monitored kinetically by measuring the NADPH decrease via its absorbance at 340nm.Results The method affords a simple,rapid,and sensitive assay with good precision and extended linearity.Test results compare closely with the HPLC precedure.The proposed method is not subject to interference from several metablites or from the drugs used commonly in clinics.Conclusion The method is easily automated and is suitable for routine work in clinical laboratories.

AIM:To detect the effect and underlying mechanism of parthenolide ( PN) on neointimal hyperpla-sia.METHODS:After 1 week of high-fat feeding, 30 male New Zealand white rabbits (2.0~2.3 kg) were randomly di-vided into 6 groups: sham +NS, rabbits received 0.9% normal saline after sham operation; sham +DMSO, rabbits re-ceived DMSO after sham operation;balloon injury(BI)+NS, rabbits received NS after balloon injury;BI+DMSO, rabbits received DMSO after balloon injury;BI+PN low, rabbits received PN at 1 mg/kg after balloon injury;BI+PN high, rab-bits received PN at 2 mg/kg after balloon injury .The drugs were intraperitoneal injected once a day after the operation until sacrifice.After fed with high-fat diet for 4 weeks, the intima-media thickness, the expression of caspase-1, IL-1β, the lev-els of IL-8, TC, TG, LDL and HDL in the serum were measured .RESULTS:Compared with sham +DMSO group, the thickness of intima, the amount of caspase-1, IL-1βand IL-8 in BI +DMSO group were significantly increased ( P <0.05).The levels of caspase-1, IL-1βand IL-8 were significantly decreased in BI +PN high group compared with BI +DMSO group (P<0.05).CONCLUSION:Neointimal hyperplasia is suppressed by PN after balloon injury , the potential mechanism may be associated with its anti-inflammatory role .

Objective To study the expression of KAI1mRNA in cervical carcinoma and its clinical significance.Methods The transcription-polymerase chain reaction(RT-PCR) method was applied to detect the expression of KAI1mRNA in 53 fresh freezing cervical tissues.The findings were compared with the clinicopathological data.Results The expression of KAI1mRNA was (0.309?0.093) in normal cervix,(1.608?0.289) in cervical intraepithelial neoplasia(CIN),(0.948?0.910) in cervical carcinoma respectively.The relative densitometric value of KAI1mRNA decreased in late stage of cervical cancer in local cervical cancer with lymphatic metastasis(P

ObjectiveTo compare the capability of optimized acellular(OA) allograft and xenograft reparing rat sciatic nerve defect by observing the immunological rejection, early functional recovery and nerve regeneration in the adult rats, which had been made a 1.0cm long gap in the continuity of the sciatic nerve.MethodsThe right sciatic nerve of adult Sprague-Dawley(SD) rats were exposed and 1.0cm long segment of the nerves were removed and repaired by fresh rabbit nerve and autofrafts, OA rat and rabbit nerve. After 1 and 3 months respectively,sciatic functional index (SFI),electrophysiological and histology studies were detected to evaluate immunological rejection,early functional recovery and nerve regeneration. ResultsThe immunological rejection, functional recovery and nerve regeneration in OA xenografts were compared with that in OA allografts, autografts and fresh allografts. One month after the surgery, the levels of CD8+ T cells and macrophages that infiltrated the grafts,the SFI and the axon density at the midpoints of them were similar within OA grafts and autografts(P ＞ 0.05),but all statistically distinguishable from fresh allografts(P ＜ 0.05).And better results were got after 2 months(P ＜ 0.05).ConclusionsThe results imply that OA xenografts is as good as OA allografts,which can be immunologically tolerated and that the removal of cellular material and preservation of the matrix are beneficial for promoting regeneration and functional recovery through an OA procedure.And this gives another promising option to repair peripheral nerve defect.

Objective To investigate the clinical application value of three-dimensional ultrasonic inspection in prenatal diagnosis.Methods 90 cases of fetal formations were Tetrospecthely analysed,which were diagnosed by two-dimensionul associtated with three-dimensional ultrasonic inspection.We compared the ultrasonic inspection with the clinical diagnosis.Results The all coincidence rate was 100%by combining the two measures,three-dimensional ultrasonic inspection can make up the defect of two-dimemional ultrasonic inspection.Conclusion Two-dimensional combined with three-dimensional ultrasonic inspection can accurately diagnose fetal formations,and increase the detection rate.

Objective To investigate the efficacy of bend-assisted laparoseopie fight bemicolectomy for right colon carcinoma in elderly obesity patients. Methods 20 obesity eases undergoing hand-assisted laparoscopic right bemicolectomy were retrospectively reviewed, and compared with 25 obesity cases with transabdominal right hemicolectomy in the same period. The safety, recovery, eradication and stress reaction were compared. Results In hand-assisted laparoscopic group, there were less bleeding loss, rapid recovery to normal temperature and gastrointestinal function,ont of bed activity, short-time hospitalization compared with transabdominal group(P <0.01). There were no differences in operative time, numbers of lymph nodes removed, postoperative complications and length of specimen between two gronps(P > 0.05). CRP, IL-6, adrenocorticotrophic hormone (ACTH) and cortisol were more significantly increased in two groups after operation than before operation (P < 0. 05) , and this increase was moreprominent in transabdominal group than in hand-assisted laparoacopic group (P < 0.05). Conclusion Hand-assis-ted laparoscopic right hemicolectomy is a safe and effective way for elderly obesity patients,especially in postoperative recovery and physical stress reaction.

Objective To investigates the gene expression of telomerase by real-time quantitative telomeric-repeat amplification protocol assay (RTQ-TRAP) in the peripheral blood mononuclear cells (PBMC) of colorectal carcinoma patients and the relationship between the telomerase gene expression in PBMC and clinicopathological features. Methods Peripheral blood samptes were collected from 71 colorectal carcinoma patients, 20 benign colorectal disease patients and 25 normal controls. The telomerase gene expression in PBMC was measured by RTQ-TRAP, and serum CEA in colorectal carcinoma patients was measured by chemiluminescence immunoassay. Results The gene expression of telomerase in PBMC was positive in 50 out of 71 cancer patients (70. 4% ) , 1 out of benign patients (5. 0% ), respectively. The difference of the telomerase gene expression of PBMC in benign colorectal diseases and cancer patients was significant (χ2 = 24. 521, P < 0. 001 ). There was no significant association between the expression of telomerase and patient's gender, age, Dukes stage, and tumor site. The positive rate of CEA in colorectal carcinoma patients was not significantly higher than positive rate of telomerase gene expression(χ2 = 2. 286,P = 0. 125). Conclusions The RTQ-TRAP method is highly accurate and sensitive in measuring telomerase gene expression. The detection of telomerase gene expression in PBMC of colorectal carcinoma patients is a simple and useful molecular marker for the diagnosis of colorectal carcinoma.

Objective To investigate the effect of nifedipine on renal ischemia-reperfusion injury (I/R) in rats and the possible mechanism. Methods Forty-two male SD rats weighing 220-250 g were randomly divided into 3 groups with 14 animals in each group: sham operation group (group S); I/R group and I/R + nifedipine group (group N). The renal ischemia was induced by occlusion of bilateral renal arteries and veins for 45 min followed by reperfusion. In group S the renal arteries and veins were only exposed but not occluded. In group N, nifedipine 0.1 mg/kg was injected via the tail vein 15 min before ischemia and 15 min before reperfusion, while the equal volume of solvent was given in group I/R. The urine was collected to detect the N-acetyl-β-D-glucosaminide (NAG) activities at 6 and 24 h of reperfusion. Arterial blood samples were taken at 6 and 24 h reperfusion for determination of concentrations of serum creatinine (Cr), malondialdehyde (MDA) and NO. The rats were then sacrificed and the kidney was removed to detect the apoptotic cells in renal cortex and expression of heat shock protein 70 (HSP70) and endothelin- 1 (ET-1). Results The concentrations of serum Cr and MDA, NAG activity, expression of HSP70 and ET-1 and apoptosis rate were significantly increased in group I/R compared with group S (P ＜0.05). The concentrations of serum Cr and MDA, NAG activity, ET-1 expression and apoptosis rate were significantly decreased, while the serum NO concentration was significantly increased in group N compared with group I/R (P ＜ 0.05). Conclusion Nifedipine can attenuate renal I/R injury and the underlying mechanism may be related to the inhibition of ET-1 expression.

AIM: To observe the neuroprotective effect of combined treatment with taurine and diazepam against focal cerebral ischemia-reperfusion in rats. METHODS: Sixty male Sprague-Dawley rats were randomly divided into five groups: sham-operation group, vehicle group, taurine group (200 mg/kg, ip), diazepam group (10 mg/kg, ip) and combined treatment group (taurine 100 mg/kg+diazepam 5 mg/kg). Focal cerebral ischemia was induced by the method of middle cerebral artery occlusion (MCAO) in rats, and reperfusion was emerged by removing the thread 2 h later. The drugs were administered respectively at the time of reperfusion, and subsequently repeated once 12 h later. The animals in vehicle group were intraperitoneally injected with isodose normal saline. The neurological deficit score, the brain water content and cerebral infarction were measured 48 h after MCAO. Other 5 group animals of focal cerebral ischemia-reperfusion (n=16 in each group) were set up as mentioned above and accepted treatments 10 h after reperfusion, likewise repeated once 12 h later. Twelve animals in each group were adopted the same management as the previous 5 groups at 48 h after MCAO. The remained 4 animals in each group were sacrificed until two weeks after MCAO to observe the histopathological changes by nissl staining. RESULTS: Compared to vehicle group, the animals in combined treatment group at 2 h or 12 h after MCAO both decreased the neurological deficit score, reduced the brain water content and infarct volume (P<0.01 or P<0.05). The combined treatment significantly alleviated the neurological necrosis as well. The neuroprotective effect of the combined treatment was superior to that of using taurine or diazepam alone. CONCLUSION: These results suggest that combination of taurine and diazepam treatment has a coordinate neuroprotective effect on both the acute and chronic brain damage of focal cerebral ischemia-reperfusion.

Objective:To determine the role of lncRNA TUG1 in pancreatic β cells functioning both in vitro and in vivo.Methods:The lncRNA TUG1 expression in mice pancreas,brain,muscle and other different tissues was examined through qRT-PCR.MTT,flow cytometry,GSIS,ELISA and immunochemistry analyses were performed to detect the effect of lncRNA TUG1 on insulin secretion in vitro and in vivo.Results:lncRNA TUG1 was highly expressed in pancreatic tissue compared with other organ tissues.Knockdown of lncRNA TUG1 expression resulted in decreased insulin secretion in β cells both in vitro and in vivo.Immunochemistry analyses showed decreased relative islet area after treatment with lncRNA TUG1 siRNA.Conclusions:Downregulation of lncRNA TUG1 expression can affect insulin secretion in pancreatic β cells in vitro and in vivo,and lncRNA TUG1 may represent a factor that regulates the function of pancreatic β cells.