Objective To determine the mechanism of nitric oxide synthase(NOS) and prostacyclin(PGI2) acting on splanchnic hyperdynamic circulation of portal hypertention(PHT). Methods Ninety-six Sprague-Dawley rats were divided into three groups, namely, intrahepatic portal hypertension(IHPH, n=31), prehepatic portal hypertension(PHPH, n=33) and sham-operated controls(SO, n=32). Animals of each group were received indomethacin(INDO) either on a short term or long term with saline as control. Portal venous pressure, together with the concentration of nitric oxide (NO) and PGI2 in serum was measured. The constitutive nitric oxide synthase(cNOS)and inducible nitric oxide synthase(iNOS)activity in the abdominal aorta and small intestine of these rats were detrmined by spectrophotometry method. RT-PCR was performed to measure the levels of iNOS and cNOS mRNA in the arteries and guts mentioned above. Results Although INDO decreased the concentration of PGI2 in serum, the long-term INDO-treated group restored splanchnic hyperdynamic circulation in both IHPH and PHPH rats, concomitant with enhanced expression of iNOS and concentration of NO(P0.05). Conclusion Overproduction of NO inducing hemodynamic abnormalities of PHT is synthesized principally by increase of iNOS. There may be a possible interaction between PGI2 and NO in hyperhemodynamics of PHT, while PGI2 may not be a mediator in the formation and development of hyperdynamic circulatory state.

Objective To observe endoplagmic reticulum stress in bone tissue of fluomsis rats and further explore the pathogenesis of skeletal fluorosis.Methods 48 Wistar rats were divided into 4 groups according to their body mass.The control and low.calcium group were fed with normal diet(0.79%calcium)and low-calcium diet(0.79%calcium)respectively,and both drank tap water(sodium fluoride concentrations＜1 mg/L).High fluoride and low.calcium plus high-fluoride groups were fed with normal diet(0.79%calcium)and low-calcium diet (0.79%calcium)respectively,and both drank tap water containing sodium fluoride(sodium fluoride concentrations 221 mg/L).During experimental period,rats were measured body mass once a week with a stand diet and water available ad libiturn.The experimental period was 3 months.The biochemical techniques were used to test the indicators of oxidative stress and ALP in seFum of fluorosis rats.The total RNA was extracted from the one side of the femur,and the transcription level of Bip,Xbp1,CHOP and PDI were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results The level of MDA in serum of low-calcium plus high-fluoride group wag higher than that of the control[(14.74±3.11)μmol/L vs(10.15±1.96)μmol/L,P＜0.05];the activity of GPx was ma~edly higher in hish-fluoride group compared with the control[(3.87±0.41)×103 U/L vs(2.85± 0.55)×103 U/L,P＜0.05];the level of uric acid in sel'um was significantly lower both in high-fluoride group and low-calcium plus high-fluoride group compared with the respective control and the low-calcium group[(73.95± 9.52)μmol/L vs(110.43±25.48)μmol/L,(54.32±22.09)μmol/L vs(101.71±17.01)μmol/L,P＜0.05]. The activity of ALP wag obviously higher in low-calcium plus high-fluoride group compared with the control [(24.77±4.57)×103U/L vs (12.91±3.97)×103U/L,P＜0.01)].The mRNA expression of Bip/GAPDH in bone tissue was markedly higher in bone of high-fluoride group and low-calcium plus high-fluoride group compared with the control(1.38±0.24,1.35±0.12 vs 1.14±0.06,P ＜ 0.05). The expression of Xbp1/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control and the low-calcium group (1.48±0.20 vs 1.02±0.25,1.07±0.25,P ＜ 0.05 or ＜ 0.01);and CHOP/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control(0.84±0.18 vs 0.52±0.07,P ＜ 0.05 ). Conclusions Accelerated osteogenetic action is seen in fluorosis rats,accompanied by oxidative stress and bone endoplasmic reticulum stress,which is likely involved in the pathogenesis of skeletal fluorosis.

Objective To explore the blood drug concentration monitoring of sustained-release valproate(DK)in children with epilepsy,focusing on the selection of sampling time and evaluation of the results.Methods Two hundred and seventy-one children taking DK and 155 children taking sodium valproate syrup(VPA Syr)were involved and their serum were taken when achieved steady state to determine the valproic acid level using fluorescence polarization immunoassay.They were divided into 4 groups,which were DK taken once daily group(DK qd group,126 children),DK taken once daily at night and sampled on morning group(DK qn group,26 children),DK taken every 12 h group(DK q12 h group,119 children),VPA Syr q12 h group(155 children).Determine the proportion of the blood drug concentration of each group below,ithin and above the therapeutic range for valproate(50-100 mg/L)were determined.The data were analyzed by t test.Results The Cmin of DK qd group were(73.09?19.91)mg/L,significantly lower from the serum concentration of DK qn and sampled on morning group [(94.94?25.44)mg/L](P0.05).Conclusions DK qn should sampled at night before the night dose.The Cmin of DK q12 h was higher according to the therapeutic range,it's favorable range still needs clinical practice.

Objective To evaluate effects of trichostain A (TSA) on cell proliferation, cell cycles, apoptosis and invasiveness of multiple myeloma cell line U266; as well as active changes of methylation regulating proteins including DNA methyl-transferase(DNMTs), methyl-binding domain (MBD) proteins: MBD2 and MeCP2 after treated with TSA. Methods U266 cells were treated with different concentrations of TSA for 12, 24, 48 and 60 h. The proliferation activity of U266 cells was detected by MTT and the IC50 of 24 h was calculated. After U266 cells were treated with IC50, cell cycles were check out by dying with PI. mRNA of matrix metalloproteinase-2(MMP-2), bc1-2, bcl-xl and methylation regulating proteins (DNMTs, MBD2 and MeCP2) were detected by real-time PCR. FCM and Western blotting were used to measure expressions of MMP-2 and MBD2. Results MTT results revealed TSA inhibited proliferation of U266 cells in a dose-and time-dependent manner and the IC50 of 24 h was 0.07 μmol/L FCM analysis showed that TSA could arrest the cell cycle in G0/G1 and the proliferation index (PI) in U266 cells [(49.90 0.39)%]were significantly different after exposed to TSA (0.7 μmnol/L for 24h compared with that in the control cells[(55.78 0.49)%](P <0.01). After treated by TSA, the 2-△△Ct of MMP-2, bcl-2 and bcl-xl were 0.71 0.06, 5.04 0.92 and 2.95 0.35, respectively. There were great changes on mRNA of DNMT, MBD2 and MeCP2. TSA could reverse the transcription of DNMT, MBD2 and MeCP2. Conclusion TSA can arrest the U266 cell cycle in GVG, to prevent its proliferation and promote apoptosis, which maybe greatly connect with the changes of the methylation regulating proteins.

Objective To explore the association between clinical pathological characteristics and the expressions of epidermal growth factor receptor (EGFR) and protein kinase B (AKT) in gastric carcinomas.Methods The expressions of EGFR and AKT were measured with immunohistochemical method in the cancer tissues and cancer-adjacent normal tissues from 153 cases of patients with gastric cancer.The association between clinical pathological characteristics and their expressions were analyzed.Results The expressions of AKT and EGFR in gastric cancer tissues had no relationship with gender,age,pathological type,and the degree of differentiation (P ＞ 0.05).A positive correlation was existed between the EGFR and TNM stages (x2 =5.43,P ＜0.05).The AKT was positively related to the size,T stage,and TNM stage of the tumor,respectively (x2 =4.73,4.95,5.32,P ＜0.05 orP ＜0.01).The levels of AKT (x2=4.83,4.75,P ＜0.05) and EGFR(x2 =4.67,4.58,P ＜0.05) in the gastric cancer tissues with lymph node and/or distant metastasis were significantly higher than the gastric cancer tissues without metastasis,respectively.Conclusions The over-expressions of AKT and EGFR would benefit the diagnosis and stages of a gastric cancer and the determination of its metastasis.

Emergencies ; Environmental Exposure ; standards ; Humans ; Maximum Allowable Concentration

Objective To prospectively assess the predictive power of centralization phenomenon in the curative effect of automated PLD.Methods The survey population was consisted of 109 patients with inclusion heraiation demonstrated by CT/MRI,74 men and 35 women with average age of 43.1 years(17～75 years). All were complained of low back pain,with varying degrees of lower extremity pain and altered sensation, lasting for more than 2 months;including one symptomatic disc in 99 patients and two symptomatic discs in 10 patients.Patients were undergone dynamic mechanical spinal test and reported whether the test would aggravate their pain.The assessment included forward flexion,extension,rotation of the trunk to the right and left, rotation to the left with fight extension,rotation to the fight with left extension,and whether straight leg raising in the supine position would aggravate back pain or leg pain.Symptom resposes were categorized into three groups:centralization group(CG),partial-centralization group(PCG)and noncentralization group(NCG). Centralization of pain is the progressive retreat of the most distal extent of the referred or radicular pain toward or to the lumbar midline.Noncentralization of pain is the peripheralization of pain in one or more directions, and no change in the distal-most pain location or intensity.All patients received a single therapy with PLD. Results A follow-up of 109 patients for 3 to 6 months,including 46 cases with 24 as exellent and 22 as good reaching 100% of excellent good rate in CG by MacNab standards;43 cases with 5 as exellent,29 as good,9 as fair and poor,with total effective rate of 79.1% in PCG.Twenty cases of NCG symptoms showed no improvement and therefore surgery was considered.Conclusions Centralization phenomenon occurrence during initial mechanical evaluation is a very accurate predictor for successful PLD outcome.Nonoccurrence of centralization would accurately predict poor PLD outcome and thus helpful as early predictor of the need for surgical treatment.

Objective To set up a system in vitro to rapidly recover plasma proteins lost during artificial-liver treatment.Methods The polyprotein precipitation was obtained by all proteins whose isoelectric point pH value were between 7.3 and 5.1,which collided with each other and aggregated using gradient pH co-precipitation(adding 1 mol/L citric acid slowly in the plasma solution to change the pH values gradually from 7.3 to 5.1 in 5 h)combined with salting out(degree of saturation of NaCl is 33%,reacted for 5.5 h at 4℃)or low-temperature ethanol precipitation(40% ethanol, reacted for 5.5 h at -7℃)so that to get rid of toxicants by discarding the supernatant.Results In the range of pH 5.1-7.3,50%(29g/57g)of the total plasma proteins had been recovered by the gradient pH salting out and 41%(25 g/61g)by the gradient pH low-temperature ethanol co-precipi- tation.The protein remained in the supernatant was mostly albumin and its combined bilirubin.The levels of total bilirubin decreased to 0.07% and 0.06% of the original levels by these two methods respectively and the serum HBV DNA level decreased to be undetected(quantitative PCR).Conclu- sions The proteins with close isoelectric point can co-precipitated with the presence of high concen- tration of NaCl or low-temperature ethanol and by changing the pH value gradually.The total protein in the discarded plasma during artificial-liver treatment can be recovered rapidly using the gradient pH coprecipitation.

A method to determine dihydroxyacetone (DHA) in fermentation broth was developed by high performance liquid chromatography (HPLC). DHA was separated on a Alltima C18(5?m,250?4.6mm). The mobile phase was 0.5% methanol solution (pH adjusted to 3.0 with H3PO4), the flow-rate was 1.0 ml/min and the detective wavelength was 200 nm. The detection limits of DHA was 0.1 g/L～10.0 g/L. 6.2 g/L DHA in the fermentation broth was detected by HPLC, which was in agreement with the result by spectrophotometric method.The method was applicable for DHA determination in the fermentation process.

More than 20 strains capable of producing dihydroxyacetone from glycerol were isolated from 4 different natural environment samples by using two detection methods. The strain 6-8 which could grow on medium containing glycerol as sole carbon source had a higher converting capability. Under a better culture, the highest DHA production of the strain 6?8 reached 6.4 g/L. In addition to general morphological and bio-chemical characteristics, the strain 6?8 was identified by 16S rDNA sequence and systematic analysis. The results showed that 16S rDNA sequence of the strain 6-8 had similarity of 99.7% with Acinetobacter sp. suggesting that the strain 6-8 is one of subspecies of Acinetobacter sp.