Objective To verify a new revised method with low usage amount of arsenic trioxide for determining urinary iodine by As(Ⅲ)-Ce4+ catalytic spectrophotometry using ammonium persulfate digestion.Methods The standard curve linearity,sample detection limit,precision and accuracy of determining urinary iodine of this modified method were verified according to Determination Methods of Chemicals in Biological Materials.Results The linear correlative coefficients of the 0-300 μg/L range and 300-1200 μg/L range calibration curve were-0.9998--1.0000(n =6) and-0.9998--1.0000,respectively.The detection limit for iodine was 1.3 μg/L.The relative standard deviations were 1.5％ (1.1/71.3)-2.5％ (6.2/244.9) when measuring 3 urine samples with iodine concentration of 71.3-244.9 μg/L,and 0.6％(2.4/388.5)-1.7％(17.3/1018.0) when measuring 3 urine samples with iodine concentration of 388.5-1018.0 μg/L,respectively(n =6).The test results of the four urinary iodine national standard materials with iodine concentration of 73.0,206.0,556.0 and 883.0 μg,/L were all within the given value range and the average value relative deviation was 1.8％ (1.3/73.0),0.4％ (0.8/206.0),0.2％ (1.0/556.0) and-1.6％(-13.7/883.0),respectively (n =6).The average recovery was 98.8％ with a range of 93.2％ (186.3/200.0)-103.4％(51.7/50.0) when measuring 3 urine samples with iodine concentration of 64.6-144.9 μg/L and 3 urine samples with iodine concentration of 346.8-574.4 μg/L,respectively.Conclusions This new modified method greatly reduces the amount of waste containing arsenic,and can directly take urine samples with high iodine concentration to digest and determine without dilution.It is performed with good standard linear curve,better precision and high accuracy,and in line with the analysis of biological samples requirements.

OBJECTIVETo observe the clinical efficacy difference between segmentation scraping and conventional acupuncture for cervical spondylosis (CS) so as to provide effective treatment method.

METHODSEighty-five cases of cervical type of CS were randomly divided into a scraping group (44 cases) and an acupuncture group (41 cases). The segmentation scraping therapy was used in the scraping group. The scraping group was treated with focusing on scraping the head and joint part of neck and occiput in the upper cervical spine injury, and focusing on scraping the lower section of cervical and shoulder in the lower cervical spine injury, once every seven days, totally for 3 times. In the acupuncture group, Fengchi (GB 20),Wangu (TE 5), Tianzhu (BL 10),Neck-Jiaji (EX-B 2), etc. were selected,once daily,for 15 days. The visual analogue scale (VAS) was used to evaluate the immediate analgesic effect after the first treatment and the clinical efficacy was observed after the end of treatment.

RESULTSAfter the first treatment, the score of VAS was decreased significantly in the scaping group (P < 0.01), but there was no significant difference in the acupuncture group compared with those before treatment (P > 0.05); the score of VAS in the scaping group after the first treatment was lower than that in the acupuncture group (3.66 +/- 0.74 vs 5.43 +/- 0.35, P < 0.01). Compared with before treatment, the scores of VAS were decreased significantly after treatment in two groups (both P < 0.01), but without significant difference between two groups (P > 0.05); the effective rate was 95.5% (42/44) in the scaping group and 87.8% (36/41) in the acupuncture group, the curative effects were similar (P > 0.05).

CONCLUSIONBoth of scraping and acupuncture therapies have good analgesic effect for cervical spondylosis, and overall effects are similar, but the immediate analgesic effect of scraping thrapy is better than that of conventional acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Neck Pain ; therapy ; Spondylosis ; therapy ; Treatment Outcome

This study was aimed to investigate the expressions of genes hoxb2 and hoxb4 after interference of the proliferation and differentiation of hematopoietic stem cells (HSC) to the erythroid progenitors (CFU-E) in vitro by using all-trans retinoic acid (ATRA). The cord blood was collected from 12 cases of fetal placenta umbilical vein and cultured by using culture technique of HSC in vitro. The proliferation and differentiation of HSC to CFU-E were interfered with 6 x 10(-8) mol/L of ATRA. The expression levels of genes hoxb2 and hoxb4 in blank control and ATRA groups were detected by FQ-RT-PCR on day 3, 7 and 10 of culture. The results showed that the expressions of genes Hoxb2 and hoxb4 were a little on day 3, obviously increased on day 7 and reached highest level on day 10 in 2 groups. The expression level of hoxb4 on day 3, 7 and 10 in blank control group was obviously higher than expression level of hoxb2. As compared with blank control group, the expressions of genes hoxb2 and hoxb4 in the ATRA group were significantly up-regulated. It is concluded that the genes hoxb2 and hoxb4 all expressed in process of proliferation and differentiation to erythroid progenitors, which suggests that hoxb2 and hoxb4 relate to erythroid hematopoiesis, and the hoxb4 has more great relevance to erythroid hematopoiesis as compared with hoxb2. The ATRA (6 x 10(-8) mol/L) can up-regulate the expression of hoxb2 and hoxb4 significantly.
Cells, Cultured ; Female ; Fetal Blood ; cytology ; Gene Expression ; drug effects ; Hematopoietic Stem Cells ; drug effects ; Homeodomain Proteins ; genetics ; Humans ; Pregnancy ; Transcription Factors ; genetics ; Tretinoin ; pharmacology

OBJECTIVETo construct a three plasmids lentiviral vector containing canine coagulation factor IX (cFIX) gene with ubiquinone promoter (PUB) and observe the expression of cFIX gene.

METHODSLentivirus was generated by transient three-plasmid transfection, namely, the VSV-G envelope expression cassette, the delta NRF packaging plasmid and the PTK 164 plasmid. Viral particles were used to infect the target cell, third passage mesenchymal stem cells (MSCs) and 293T cell respectively at MOI 3: 1. The cFIX activity was detected in cultured cells with one-stage clotting assay.

RESULTSThe MSCs were obtained in vitro. The lentivirus infected MSCs and 293T cells all expressed the active factor IX with the activity of (26.30 +/- 2.10)% and (19.70 +/- 1.53)%, respectively, which are significantly higher than that of control (1.00 +/- 0.05)%.

CONCLUSIONSThe lentiviral vector of three plasmids with ubiquinone promoter (PUB) was constructed and can transfect the MSCs and 293T cells.

Animals ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; Dogs ; Factor IX ; genetics ; metabolism ; Genetic Vectors ; Hemophilia B ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; Plasmids ; genetics ; Transfection

OBJECTIVE: To evaluate influences of regular-dose of adriamycin (ADR) on heart function and sarcoplasmic reticulum (SR) Ca2+ -ATPase in cardiomyocyte of rabbits. METHODS: Nine rabbits received intraveneous injection of ADR (2mg/kg) once a week for 8 weeks, the rabbits injected with saline were used as control group. Cardiac output (CO), blood pressure (BP), mean artery pressure (MAP), left ventricular systolic pressure( LVSP), left ventricular end-diastolic pressure (LVEDP), calcium in cardiomyocyte (MyoCa2+) of rabbits and SR Ca2+ -ATPase activity were examinated 3 weeks after the final injection. RESULTS: CO, LVSP and SR Ca2+ -ATPase activity were significantly decreased in ADR treated group compared with the control group. Conversely, LVEDP and MyoCa2+ were significantly increased in ADR treated rabbits. CONCLUSION: Heart function can be decreased by regular-dose of ADR in injection. Calcium overload in cardiomyocyte and decrease of SR Ca2+ -ATPase activity is important physiopathologic mechanism in ADR-induced impairment of heart.

After oral administration of low, middle, high dose of simulative Maijunan tablets to SD rats and puerarin, hydrochlorothiazide at middle dosage to SD rats separately, plasma samples were collected at different times and treated with acetonitrile to precipitate protein. The contents of puerarin and hydrochlorothiazide in plasma were determined by HPLC method. The mean plasma concentration-time profiles of puerarin and hydrochlorothiazide at different dosages of medication administration were processed by 3P97 pharmacokinetic software and SPSS statistics 17.0 software. The results indicated that the in vivo kinetic processes of puerarin in rats were all fitted to a two-compartment open model and hydrochlorothiazide fitted to a one-compartment open model. Hydrochlorothiazide in vivo kinetic process in rats was in accordance with the linear dynamics. The combination of hydrochlorothiazide and rhynchophylla with pueraria promoted the absorption, reduced the elimination rate and prolonged the action time of puerarin in vivo. Meanwhile, the combination also promoted the absorption rate and the bioavailability, prolonged the action time and the accumulation time of hydrochlorothiazide in vivo.
Animals ; Biological Availability ; Drugs, Chinese Herbal ; chemistry ; Female ; Hydrochlorothiazide ; pharmacokinetics ; Isoflavones ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tablets

Animals ; Cell Proliferation ; drug effects ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mesangial Cells ; cytology ; drug effects ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; genetics

Objective To observe the efficacy difference between heavy-dose moxibustion at Sha-petechiae and ordinary suspended moxibustion in treating neck-type cervical spondylosis due to cold coagulation and blood stagnation.Method A total of 113 patients with neck-type cervical spondylosis due to cold coagulation and blood stagnation were randomized into a treatment group of 58 cases and a control group of 55 cases.The treatment group was intervened by heavy-dose moxibustion at Sha-petechiae after scraping therapy;the control group was treated with conventional moxibustion therapy.The cervical symptoms and signs were evaluated by using LU's scale scores.The improvement in symptoms and clinical efficacy were observed after the intervention and half a year later.Result The Visual Analogue Scale (VAS) and LU's scale scores were significantly changed after the intervention and half a year later compared with those before the intervention in both groups (P＜0.01).After the intervention and half a year later,there were significant differences in comparing the VAS and LU's scale scores between the two groups (P＜0.01).The total effective rate and markedly effective rate were respectively 94.8％ and 77.6％ in the treatment group versus 76.4％ and 52.7％ in the control group,and the between-group differences were statistically significant (P＜0.01).Conclusion Heavy-dose moxibustion at Sha-petechiae is an effective approach in treating neck-type cervical spondylosis due to cold coagulation and blood stagnation,and its short-term and long-term efficacies are both superior to those of ordinary moxibustion.

OBJECTIVEThis study examined the effect of recombinant human erythropoietin (r-HuEPO) on the serum levels of neuron-specific enolase (NSE), S-100β protein and myelin basic protein (MBP) in young rats 24 hrs after lithium-pilocarpine-induced status epilepticus (SE) in order to study the potential role of r-HuEPO in epileptic brain damage.

METHODSForty 19-21-day-old male Sprague-Dawley (SD) rats were randomly divided into four groups (n=10): normal control group, SE, r-HuEPO pretreated-SE and r-HuEPO. SE was induced by lithium-pilocarpine. R-HuEPO (500 IU/kg) was intraperitoneally injected in the r-HuEPO pretreated-SE and r-HuEPO groups 4 hrs before SE. Serum levels of NSE, S-100β and MBP were determined 24 hrs after the SE event.

RESULTSSerum levels of NSE, S-100β and MBP in the SE group increased significantly compared with those in the normal control and the r-HuEPO groups (P＜0.05). The r-HuEPO pretreated-SE group showed significantly decreased serum levels of NSE, S-100β and MBP compared with the SE group (P＜0.05).

CONCLUSIONSr-HuEPO may reduce the expression of NSE, S-100β and MBP and thus might provide an early protective effect against epileptic brain injury.

Animals ; Erythropoietin ; pharmacology ; therapeutic use ; Male ; Myelin Basic Protein ; blood ; Nerve Growth Factors ; blood ; Phosphopyruvate Hydratase ; blood ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; blood ; Status Epilepticus ; blood ; drug therapy

In the present study, we investigated the inhibitory action of ketanserin on wild-type (WT) and Y652 mutant human ether-a-go-go-related gene (HERG) potassium channels expressed in Xenopus oocytes and the effects of changing the channel molecular determinants characteristics on the blockade with and without ketanserin intervention using standard two-microelectrode voltage-clamp techniques. Point mutations were introduced into HERG gene (Y652A and Y652R) and subcloned into the pSP64 plasmid expression vector. Complementary RNAs for injection into oocytes were prepared with SP6 Cap-Scribe after linearization of the expression construct with EcoR I. Clampfit 9.2 software was employed for data collection and analysis. Origin 6.0 software was used to fit the data, calculate time constants and plot histograms. The results showed that ketanserin blocked WT HERG currents in voltage- and concentration-dependent manner and showed minimal tonic blockade of HERG current evaluated by the envelope of tails test. The IC50 value was (0.38+/-0.04) micromol/L for WT HERG potassium channel. The peaks of the I-V relationship for HERG channel suggested a negative shift in the voltage-dependence of activation after using ketanserin, whose midpoint of activation values (V1/2) were (-16.59+/-1.01) mV (control) vs (-20.59+/-0.87) mV (ketanserin) at 0.1 micromol/L, (-22.39+/-0.94) mV at 1 micromol/L, (-23.51+/-0.91) mV at 10 micromol/L, respectively (P<0.05, n=6). Characteristics of blockade were consistent with an open-state channel blockade, because the extent and rate of onset of blockade was voltage-dependent, increasing at more potentials even in the condition of leftward shift of activation curve. Meanwhile, in the different depolarization duration, the fractional blockade of end-pulse step current and peak tail current at 100 ms duration was significantly lower than that at 400 ms and 700 ms, which indicated that following the channel activation fractional blockade was enhanced by the activated channels. Ketanserin could also modulate the inactivation of HERG channel, which shifted the voltage-dependence of WT HERG channel inactivation curve from (-51.71+/-2.15) mV to (-80.76+/-14.98) mV (P<0.05, n=4). The S6 mutation, Y652A and Y652R, significantly attenuated the blockade by ketanserin. The IC50 value were (27.13+/-9.40) micromol/L and (20.20+/-2.80) micromol/L, respectively, increased by approximately 72-fold for Y652A and 53-fold for Y652R compared to that of WT HERG channel blockade [(0.38+/-0.04) micromol/L]. However, between the inhibitory effects of Y652A and Y652R, there was no significant difference. In conclusion, ketanserin blocks WT HERG currents in voltage- and concentration-dependent manner and preferentially blocks open-state HERG channels. Tyr-652 is one of the critical residues in the ketanserin-binding sites.
Animals ; Ether-A-Go-Go Potassium Channels ; antagonists & inhibitors ; Humans ; Ketanserin ; pharmacology ; Mutation ; Oocytes ; Patch-Clamp Techniques ; Potassium Channel Blockers ; pharmacology ; Xenopus