A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.

To control the quality of Humulus scandens, the quality standard was established in this study. According to the method recorded in the Appendix of Chinese Pharmacopoeia (2010 Edition) , the water and ash inspections were carried out. The component luteoloside and cosmosiin in Humulus scandens were identified and assayed by TLC and HPLC. The results showed a strong characteristics microscopic of Humulus scandens, and trichoromethane-methanol-formic acid (10: 3: 0. 3) as the mobile phase of TLC, the spots at 365 nm with a UV lamp was clear. The 16 batches of samples were analyzed by HPLC with a gradient elution of acetonitrile and phosphate solution (0.2%) at a flow rate of 1.0 mL · min(-1) and detected at 350 nm. The content of luteoloside was 0.015%- 0.651% (average 0.148%); the content of cosmosiin was 0.003%-0.118% (average 0.036%). The linear calibration curve of luteoloside and cosmosiin was acquired in the ranges of 0.011-0.364 g · L(-1) (r = 1.000 0) and 0.003-0.096 g · L(-1) (r = 1.000 0), respectively. The average recovery was 100.5% and 98.5%, respectively. The methods are convenient and reliable, which can be ap- plied for quality assessment of Humulus scandens.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; standards ; Humulus ; anatomy & histology ; chemistry ; Quality Control

Objectiye To optimize the depth of the microchannel and the time point for sperm collection,and improve the efficiency of sperm screening on a microfluidic device. Methods Microchannels with four different depths of 25, 50, 100 and 200 μm were tested. Mice sperm were added to the inlet of the microchannel. The relative quantity and motility of sperm in the outlet were recorded at different collection times, i.e. ,5, 15, 30 and 60 min. Statistical method one-way ANOVA and appropriate post-hoc testing were applied to analyze differences between different groups, and further to select the best-fit depth of the microchannel and the time point for collection. Results In microchannels with depths of 25, 50, 100 and 200 μm, the sperm motilities measured in each outlet were (85.4 ± 2.3)%, (85.8 ± 5.8)%,( 87. 2 ± 2. 8 ) %, (76. 5 ± 2. 8 ) % respectively with statistical significance ( F = 5.8, P ＜ 0. 05 ). No obvious differences were found among 25-100 μm channels, however the motility dramatically decreased in the 200 μm group. The relative sperm quantities were (5.2 ±2.0)%, (7.2 ±2.5)%,(12.3 ±2.0)%,(7. 7 ± 1.1 ) % respectively with statistical significance ( F = 6. 9, P ＜ 0. 05), which increased with channel depth from 25 to 100 μm,while it decreased in the 200 μm channel Taking 2 indexes into account, 100 μmwas the most fit channel depth for sperm motility screening. The sperm motility in the outlet gradually decreased with time. At the time points of 5, 15, 30 and 60 min after adding sperm, the sperm motilities were (99. 6 ±0. 7)%, (87.2 ±2. 8)%, (79. 3 ±2. 2)% and (62. 6 ±8.0)% respectively with statistical significance ( F = 37. 3, P ＜ 0. 01 ). Yet the relative quantities of sperm in the outlet increased almost three times in this process. At the time points mentioned above, the relative quantities of sperm were (5.8±1.1)%, (10.6 ± 0.9)%, (12.1 ± 1.7)%, (17.9 ± 3.4)% respectively with statistical significance ( F = 17.8, P ＜ 0. 01 ). Thus 15-30 min was the ideal screening time. Conclusion An effective microdevice for sperm screening with optimized depth and collection time period is developed,which may contribute significantly for the screening of healthy sperm on microfluidic chips.

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.
Enzyme Inhibitors ; pharmacology ; Free Radical Scavengers ; pharmacology ; High-Throughput Screening Assays ; Superoxides ; Uric Acid ; Xanthine ; Xanthine Oxidase ; antagonists & inhibitors

To promote bilingual teaching reform and explore a proper bilingual teaching mode in China,we studied the bilingual teaching of the course “Biochemistry on special subjects”.The present paper mainly designs and discusses the object of the course,teaching materials,contents and methods as well as the building of feedback and evaluation system of the course.

OBJECTIVETo observe the pathological changes of renal tissue in the rats with paraquat (PQ) poisoning as well as the serum creatinine (SCr) levels and expression levels of hypoxia-inducible factor-1α (HIF-1α) and transforming growth factor-β (TGF-bgr;) in renal tissue at different time points after PQ poisoning, and to investigate the association of HIF-1α with renal injury after PQ poisoning.

METHODSForty-eight healthy male Sprague-Dawley rats were randomly divided into control group (n = 6) and PQ group (n = 42). The control group was given a single dose of 1 ml saline by gavage; the PQ group was given a single dose of 1 ml PQ (50 mg/kg), which was prepared by diluting 20% raw liquid of PQ with saline, by gavage. The PQ group was further divided into 2, 6, 12, 24, 48, 72 and 120 h PQ subgroups (n = 6 for each subgroup) to be examined at 2, 6, 12, 24, 48, 72, or 120 h after gavage. Their arterial blood was collected for blood gas analysis as well as blood lactic acid (BLA) and SCr measurement; renal sections were subjected to HE staining; the protein expression of HIF-1α and TGF-β in renal tissue was measured by Western blot.

RESULTSThe BLA level and SCr level began to rise at 6h after poisoning. Compared with the control group, the 6, 12, 24, 48, 72 and 120 h PQ subgroups had significantly increased BLA and SCr levels (P < 0.05); the 72 and 120 h PQ subgroup showed hypoxemia (P < 0.05). The protein expression of HIF-1α in PQ group increased significantly at 6h and reached the peak level at 72 h, with a significant difference from that in the control group at 6, 12, 24, 48, 72, and 120 h (P < 0.05). The protein expression of TGF-β in PQ group began to rise at 24 h, reached the peak level at 72 h, and declined at 120 h, with a significant difference from that in the control group at 24, 48, and 72 h (P < 0.05). The protein expression of HIF-1α was positively correlated with SCr level (r = 0.9308, P = 0.0008), uncorrelated with arterial partial pressure of oxygen (r = -0.6996, P = 0.0534), and positively correlated with BLA level (r = 0.9483, P = 0.0003). The pathological changes of renal tissue mainly included the degeneration and necrosis of renal tubular epithelial cells, which worsened as the time went on and appeared less severe at 120 h.

CONCLUSIONThe HIF-1α expression in renal tissue increases significantly in the early stage of PQ poisoning, which is associated with increased BLA and SCr levels and causes upregulated expression of TGF-β that promotes renal fibrosis.

Animals ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Kidney ; metabolism ; pathology ; Male ; Paraquat ; toxicity ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; metabolism

Ischemic stroke is a primary cause of death and long-term disability all over the world. This disease is resulted from ischemia and hypoxia in brain tissues because of insufficient blood supply and causes a series of physiochemical metabolism disorders and physiological dysfunction. Its high disability ratio has bright huge burdens to society, governments and families. However, there is not efficacious medicine to treat it. In this study, a right middle cerebral artery occlusion was established in rats to observe the multi-path and multi-aspect intervention effects of Tibetan patent medicine Ruyi Zhenbao pills in reducing injuries to Nissl bodies, cerebral edema and inflammatory reactions and preventing cellular apoptosis, in order to lay a foundation for defining its therapeutic mechanism in acute ischemic stroke.
Animals ; Brain Ischemia ; drug therapy ; Male ; Medicine, Chinese Traditional ; Medicine, Tibetan Traditional ; NF-kappa B ; physiology ; Patents as Topic ; Rats ; Rats, Sprague-Dawley ; Stroke ; drug therapy

Objective To detect the expressions of CD19 and CD20 of B-lymphocytes (CD19+B, CD20+B) and natural killer (NK) cells in the peripheral blood of epileptic children and explore their significances. Methods Four hundred and fifty-eight epileptic children, admitted to our hospital from January 2008 to December 2010, were chosen as patient group; another 52 healthy subjects were chosen as controls. The expressions of CD19+B and CD20+B and NK cells were detected by flow cytometry; their results were compared. Ninety-two epileptic children were treated with intravenous immunoglobulin (IVIG) and the effect of IVIG therapy was studied. Results The CD19+B level ([22.35±6.54]％) and CD20+B level ([21.50±8.41]％) in the epileptic children were obviously higher than those in the healthy controls ([16.86±4.02)％,[16.13±4.19]％, P＜0.05); and the level of NK cells ([9.1 1±4.90]％) in the epileptic children was significantly decreased as compared with that in the healthy controls ([14.72±4.15]％, P＜0.05). The CD19+B level ([18.26±5.03]％) and CD20+B level ([16.74±5.12]％) 6months after MG treatment were decreased significantly as compared with those before treatment ([22.74±6.25]％,[21.61±8.03]％, P＜0.05]; while the level of NK cells ([14.65±4.58]％) 6 months after IVIG treatment was increased significantly as compared with that before treatment ([9.07±4.76]％,P＜0.05). Among the 92 patients treated with IVIG, 70 enjoyed good results and 22 had non-effective resutls; The changesofCD19+Blevel ([7.99±5.34]％) and CD20+B ([8.21±5.21]％) before and after treatment in effectively treated patients by IVIG were significantly different as compared with those in ineffectively treated patients by MG ([3.78±2.76]％,[3.66±2.48]％, P＜0.05); however, the changes of level of NK cells before and after treatment showed no significant difference between effectively treated patients and ineffectively treated patients ([5.28±4.55]％,[4.53％±4.43]％, P＞0.05). Conclusion Dysfunctions of B-lymphocytes and NK cells exist in epileptic children, and MG treatment shows good effect on immune dysfunction of them. The levels of CD19-B and CD20-B can be used as monitoring indicators in the IVIG treatment of epileptic children.

The detailed operating methods and the clinical application of the unique acupuncture therapy established by professor XIE Qiang and applied specially in otorhinolaryngologic department are introduced, ie. acupuncture and moxibustion methods for regulating channels and contacting qi, transferring focus of excitation, needling movement and needling ying combined with minimally invasive needle scalpel. Meanwhile, the position of the experience acupoints named Kaiyin 1 (ease-up the voice 1) and Kaiyin 2 (ease-up the voice 2) and their manipulation are recommended, and the clinical application of these acupuncture methods and acupoints for inflammation and pain in otorhinolaryngologic department are explained.
Acupuncture Therapy ; Adult ; Humans ; Male ; Middle Aged ; Moxibustion ; Otorhinolaryngologic Diseases ; therapy ; Pain Management ; Young Adult

OBJECTIVETo investigate the presence of abnormal metabolism in the thalamus and hypothalamus in patients with first-episode depression.

METHODSThirty drug-naive patients with first-episode depression and 30 age-matched controls were scanned with proton magnetic resonance spectroscopy ((1)H-MRS) for Naa, Cho, Cr and mI.

RESULTSCompared with the control group, the patients showed significantly reduced mI and mI/Cr of the hypothalamus, reduced mI/Cr of the left thalamus, and lowered Cho, ml, and ml/Cr of the right thalamus (P<0.05).

CONCLUSIONPatients with first-episode depression may have myo-inositol and phosphoric acid metabolism disorder in the thalamus and hypothalamus with malfunction of cellular osmotic pressure adjustment mechanism. Abnormal mI/Cr in the thalamus and hypothalamus may represent an important biochemical change in advanced patients with depression.

Adolescent ; Adult ; Case-Control Studies ; Choline ; metabolism ; Creatine ; metabolism ; Depression ; diagnosis ; Female ; Humans ; Hypothalamus ; metabolism ; Inositol ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Male ; Middle Aged ; Protons ; Thalamus ; metabolism ; Young Adult