Objective Study on the traceability of serum enzyme ass ays by testing enzyme reference material in clinical laboratories. Methods 50 laboratories were involved in this survey. One enzyme reference material was send to each participate lab. The reference material was tested by use of routine method and the results were recorded. All lab data were processed with computer. Results Compare with the target values, the bias of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK) and lactate dehydrogenase (LD) assays were 6 3%, 5 5%, -5 9% and -5 0% respectively, the bias of alkaline phosphatase (ALP) assays was -36 6. The inter laboratory coefficient variation of the amylase assay was 29 2%. Conclusions The results of enzyme assay in clinical laboratories could be traced to the international reference material.

Objective Investigating for the standardization of serum enzyme determination by using enzyme calibrator in lab tests Methods 150 laboratories were involved in this program. One enzyme calibrator and three patient samples were send to each participate lab The calibrator was tested using routine method and record the results The patient samples were analyzed before and after calibration Record the results All lab data was processed with computer Results Compare with the target values, the bias of alanine aminotransferase (ALT), aspartate aminotransferase (AST), amylase (AMY) and lactate dehydrogenase (LD) assays were 3 8%, -1 8%, 2 3% and -5 2% respectively, the bias of alkaline phosphatase (ALP), gamma glutamyltransferase (GGT) and creatine kinase (CK) assays were 13 7%, -13 9 and -19 2% respectively The precision of the enzyme assays were improved by using calibrators Conclusions The traceability of ALT, AST, AMY and LD assays were reasonable Calibrating by using enzyme calibrator can improve the assay precision among laboratories

Objective Understand the variation of serum creatinine measurement in clinical laboratories of some hospitals in Beijing.Methods 8 samples of mixed frozen human serum added different creatinine concentration standard materials(the creatinine concentration were80-1 000 ?mol/L)and 8 samples of mixed frozen serum of patients(contained different creatinine concentration)were distributed to 13 clinical laboratories(31 series of detection systems)with the way of spot investigation.Every clinical laboratories measured the samples followed the standard operating procedure.Results As to the mixed frozen human serum added different creatinine standard materials,the CV of different detection systems results were 5.74%-9.68%;as to the mixed frozen patients' serum,the CV was 5.90%-11.69%. Compared with Beckman closed detection systems,the results of Dade systems(which used the kinetic alkaline pieric acid method)showed the bias were-5.99%-0.35%,and as to the other systems which measured by alkaline picric acid method,when creatinine concentrations were 200 ?mol/L,the results showed negative bias,and the greatest bias was-8.45%.The bias plots revealed negative for all of the detection systems with enzymatic method over the whole concentration range,and the greatest bias was -8.88%.Conclusions The creatinine determination results of Beckman and Dade closed detection systems were consistent.The results of detection systems which used enzymatic method were generally lower than Beckman detection systems.What's more,the creatinine measurement variations of clinical laboratories were very large,especially for the results of unclosed detection systems,so it was urgent need to solve the standardization of creatinine measurement.

Objective To explore the expression and localization of aquaporin 11(AQP11) in human term pregnancies with oligihydramnios, and its role in amniotic fluid balance.Methods We studied 55 patients who underwent elective cesarean sections, consisting of 25 patients with isolated oligohydramnios and 30 with normal amniotic fluid volume.Immunohistochemistry and reverse transcription polymerase chain reaction were employed to determine the expression and localization of AQP11 in the amnion, chorion and placenta.Results AQP11 protein was detected in expressed in the full-term pregnant women`s amnion, chorion and placenta.The expression in the amnion was positively correlated with amniotic fluid amount;the expression in the placenta was negatively correlated with amniotic fluid amount.The expression increased in the chorion with different amniotic fluid amount.Conclusion AQP11 plays an important role in regulating amniotic fluid balance.

Objective To obtain acquired immunity evidence in hamsters elicited by third stage hookworm larvae of Necator americanas(NaL3).morphology changes of NaL3 and inflammatory responses in the skin and undedying subcutaneous tissue and muscles of hamsters were observed.Methods Hamsters were immunized subcutaneously with one dose of 150 NaL3 at 2 weeks earlier,and then challenged pereutaneously with 900 NaL3.Skins were excised from post-challenge hamsters at 6,24,72 hours and 1,2 weeks,and then examined under light microscopy.Non-immunized hamsters served as negative controls.Results In non-immunized hamsters the number of NaL3 were 15,33,11.0 and 0 at 6,24,72 hours and 1,2 weeks post-infection.No damaged or dead NaL3 section was observed.All NaL3 exhibited no structural damage and infihrating inflammatory cells were absent from the sunDunding tissues.There were no cutaneous changes.In contrast.the total number of Nak sections in the skin of immunized hamsters were 25,53,15,5 and 4 at 6,24,72 hours and 1,2 weeks post-challenge.Among these NaL3 sections,damaged and dead section number were 0,24,6,0,0 and 0,0,7,5,4.At 24 hours post-challenge the Nak exhibited cutieular swelling and damage.By 72 hours post-challenge pyknosis of the somatic cells nuclei and sparseness or loss of definition in the internal structures of NaL3 were seen.One or two weeks after chanenge,the NaL3 showed severe damage or even dead with remnants.Inflammatory responses including macrophages,epithelioid cells and eosinophils infiltrating and granulomata forming were mainly seen around the NaL3 sections in the skin of immunized hamsters.Conclusions Hamsters initially immunized with NaL3 exhibited obvious acquired immunity protection against percutaneously challenged infection as evidenced by vigorous inflammatory responses in the skin and underlying subcutaneous tissue and muscle.

The culture medium and cultural conditions of solid-state fermentation of Streptomyces Menmyco-93-63 were tested in this study. The suitable medium which contains rice, sorghum, millet bran, and rice hull with the proportion of 2:2:3:3 was developed for the spore production of Streptomyces Men-myco-93-63 using single substrate screening, mixture substrate screening and orthogonal experiments, and the sporulation was up to 2.52?109 CFU/g. And then, initial charge, initial ratio of water to solid, inoculating quantity, and culture temperature impact to sporulation of Streptomyces Men-myco-93-63 were tested. The favorite cultural conditions are developed as the following: the initial charge is 15 g in 500 mL Erlenmeyer flask; initial ratio of water to solid is 1.7:1.0 (V/W, rice hull excluding), inoculating quantity is 7 mL, culture temperature is 28℃.

AIMTo study the dynamic and long-lasting expression of thrombin receptor after acute intracerebral hemorrhage (ICH) in rats.

METHODS36 rats were randomly divided into 6 groups (n = 6): Normal group and ICH model groups at 6 hours, 24 hours, 3 days, 7 days and 14 days. ICH models were produced with the induction of collagenase type VII-S. Immunohistochemical method was used to detect PAR-1 protein and RT-PCR technique was used to detect PAR-1mRNA in brain tissue around the haematoma in different groups.

RESULTSPAR-1 protein and mRNA were mild positive in normal group. In model groups, intensity of PAR-1 expression started to enhance at 6 hours, and enhanced more at 24 hours. PAR-1 expression reached the peak at 3 days and began to descend. At 7 days the descent was obvious and there was further descent at 14 days. At each time point, the PAR-1 protein positive cell number and PAR-1mRNA absorbance ratio in ICH model groups were significantly higher than those in normal group (P < 0.05 or P < 0.01). In addition, PAR-1 proteins were obviously expressed in vivo in brain capillary endothelial cell.

CONCLUSIONFunctional PAR-1 exists in brain capillary endothelial cells. Activation of PAR-1 after ICH due to the stimulation of thrombin is not only the initiating agent of cerebral edema after ICH, but also participates the development of cerebral edema.

Animals ; Cerebral Hemorrhage ; metabolism ; Disease Models, Animal ; Male ; Rats ; Rats, Wistar ; Receptor, PAR-1 ; metabolism ; Thrombin ; metabolism

The aim of this study was to observe and compare the endogenous circadian rhythm and photoresponse of Clock gene transcription in the suprachiasmatic nucleus (SCN) and pineal gland (PG) of rats. With free access to food and water in special darkrooms, Sprague-Dawley rats were housed under the light regime of constant darkness (DD) for 8 weeks (n=36) or 12 hour-light: 12 hour-dark cycle (LD) for 4 weeks (n=36), respectively. Then, their SCN and PG were dissected out every 4 h in a circadian day, 6 rats at each time (n=6). All animal treatments and sampling during the dark phases were conducted under red dim light (<0.1 lux). The total RNA was extracted from each sample and the semi-quantitative RT-PCR was used to determine the temporal mRNA changes of Clock gene in the SCN and PG at different circadian times (CT) or zeitgeber times (ZT). The grayness ratio of Clock/H3.3 bands was served as the relative estimation of Clock gene expression. The experimental data were analyzed by the Cosine method and the Clock Lab software to fit original results measured at 6 time points and to simulate a circadian rhythmic curve which was then examined for statistical difference by the amplitude F test. The main results are as follows: (1) The mRNA levels of Clock gene in the SCN under DD regime displayed the circadian oscillation (P<0.05). The endogenous rhythmic profiles of Clock gene transcription in the PG were similar to those in the SCN (P>0.05) throughout the day with the peak at the subjective night (CT15 in the SCN or CT18 in the PG) and the trough during the subjective day (CT3 in the SCN or CT6 in the PG). (2) Clock gene transcription in the SCN under LD cycle also showed the circadian oscillation (P<0.05), and the rhythmic profile was anti-phasic to that under DD condition (P<0.05). The amplitude and the mRNA level at the peak of Clock gene transcription in the SCN under LD were significantly increased compared with that under DD (P<0.05), while the value of corresponding rhythmic parameters in the PG under LD were remarkably decreased (P<0.05). (3) Under LD cycle, the circadian profiles of Clock gene transcription induced by light in the PG were quite different from those in the SCN (P<0.05). Their Clock transcription rhythms were anti-phasic, i.e., showing peaks at the light phase ZT10 in the SCN or at the dark time ZT17 in the PG and troughs during the dark time ZT22 in the SCN or during the light phase ZT5 in the PG. The findings of the present study indicate a synchronous endogenous nature of the Clock gene circadian transcriptions in the SCN and PG, and different roles of light regime in modulating the circadian transcriptions of Clock gene in these two central nuclei.
Animals ; CLOCK Proteins ; genetics ; Circadian Rhythm ; physiology ; Male ; Photoreceptor Cells, Vertebrate ; physiology ; Pineal Gland ; physiology ; Rats ; Rats, Sprague-Dawley ; Suprachiasmatic Nucleus ; physiology ; Transcription, Genetic

OBJECTIVETo explore the protective effect of baicalin against rotenone-induced injury on PC12 cells, and the po-tential mechanism of action action was also explored.

METHODPC12 cells were injured by rotenone and were treated with different concentrations (0.1, 1, 10 μmol x L(-1)) of baicalin at the same time. Cell viability was analyzed by MTT, and morphology was observed by phase-contrast microscopy. The cell apoptosis was detected by flow cytometry by Annexin V-FITC/PI staining. The intracellular ROS level was determined by fluorescence microscope with DCF-DA staining. The expression of Bcl-2, Bax and Caspase-3 was analyzed by Western blot.

RESULTThe viability of PC12 cells exposure to rotenone for 24 hour was gradually decreased with dose escalating and 1.5 μmol x L was adopted to do the following experiment. Baicalin increased cell viability, improved cell morphology and decreased intracellular ROS level. Moreover, FACS indicated baicalin attenuated the apoptosis induced by rotenone significantly. Western blot showed that Bcl-2, Bax and Caspase-3 expression in rotenone-induced PC12 cells was reversed by baicalin.

CONCLUSIONThis study has demonstrated that baicalin protects PC12 cells against rotenone-induced apoptosis, at least in part, by scavenging excessive ROS and inhibiting the mitochondrion-dependent apoptotic pathway.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Cytoprotection ; drug effects ; Flavonoids ; pharmacology ; Gene Expression Regulation ; drug effects ; Intracellular Space ; drug effects ; metabolism ; PC12 Cells ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; Rotenone ; pharmacology ; bcl-2-Associated X Protein ; metabolism

This study was to investigate the circadian rhythms and light responses of Clock gene and arylalkylamine N-acetyltransferase (NAT) gene expressions in the rat pineal gland under the 12 h-light : 12 h-dark cycle condition (LD) and constant darkness (DD). Sprague-Dawley rats housed under the light regime of LD (n=36) for 4 weeks and of DD (n=36) for 8 weeks were sampled for the pineal gland once a group (n=6) every 4 h in a circadian day. The total RNA was extracted from each sample and the semiquantitative reverse transcription polymerase chain reaction (RT-PCR) was used to determine the temporal changes in mRNA levels of Clock and NAT genes during different circadian times or zeitgeber times. The data were analysed by the cosine function software, Clock Lab software and the amplitude F test was used to reveal the circadian rhythm. The main results obtained are as follows. (1) In DD or LD condition, both of Clock and NAT genes mRNA levels in the pineal gland showed robust circadian oscillation (P< 0.05) with the peak at the subjective night or at night-time. (2) In comparison with DD regime, the amplitudes and the mRNA levels at peaks of Clock and NAT genes expressions in LD in the pineal gland were significantly reduced (P< 0.05). (3) In DD or LD condition, the circadian expressions of NAT gene were similar in pattern to those of Clock gene in the pineal gland (P> 0.05). These findings suggest that the expressions of Clock and NAT genes in the pineal gland not only show remarkably synchronous endogenous circadian rhythmic changes, but also response to the ambient light signal in a reduced manner.
Animals ; Arylalkylamine N-Acetyltransferase ; genetics ; metabolism ; CLOCK Proteins ; genetics ; metabolism ; Circadian Rhythm ; Light ; Male ; Pineal Gland ; enzymology ; metabolism ; RNA ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley