Objective:To establish an HPLC method for the determination of gallic acid,quercetin and kaempferol in Gongyanping capsules. Methods:HPLC was applied with the chromatographic conditions as follows: The chromatographic column was Agilent-SB C18 (250 mm × 4. 6 mm,5 μm) at 30℃; the mobile phrase was methanol-0. 3% phosphoric acid solution with gradient elution; the flow rate was 1. 0 ml·min-1. Results: The linearity relationship of gallic acid, quercetin and kaempferol was within the range of 98.200-491.00 μg·ml-1(r=0.999 9), 7.520-37.600 μg·ml-1(r=0.999 9) and 4.940-24.700 μg·ml-1(r=0.999 9), re-spectively;the average recovery was 96. 74%(RSD=1. 33%), 98. 18%(RSD=1. 70%) and 97. 04%(RSD=1. 28%),respective-ly. Conclusion:The method is simple, accurate and repeatable.

Objective:To study the quality standard for Herba Desmodii,and provide the scientific basis for a reasonable method of quality control. Methods:With the methods of microscopic identification,thin-layer chromatography and high performance liquid chromatography for the qualitative and quantitative determination of Herba Desmodii,the ethanol soluble extracts,moisture content and total ashes of Herba Desmodii were determined. Results:The characteristics of thin-layer chromatography were obvious;a good lineari-ty of quercetin and kaempferol was within the range of 0. 002-0. 050 μg(r=1. 000 0)and 0. 004-0. 100 μg(r=0. 999 9)with the av-erage recovery of 97.89%(RSD=0.89%,n=6)and 98.93%(RSD=1.31%,n=6),respectively. Conclusion:The method is simple,accurate and reproducible,which can effectively control the quality of Herba Desmodii.

Objective: To discuss the quality control method and index of traditional She medicine -Melastoma dodecandrum. Methods:The contents of gallic acid , ellagic acid and total flavonoids ( the total amount of quercetin and kaempferol) were deter-mined by HPLC. Water content, acid insoluble ash content, extract, heavy metals and harmful elements in 26 batches of herbs were also detected. Results:The content range of gallic acid, ellagic acid and total flavonoids was 9. 84-26. 31 mg·g-1 , 4. 13-16. 11 mg· g-1 and 1. 31-38. 28 mg · g-1 , respectively. The content range of water, acid insoluble ash and extract was 6. 70%-13. 90%, 1. 05%-6. 20% and 18. 00%-40. 80%, respectively. The content of Pb in all the samples was more than 5 ppm, the number of sam-ples with the content of Cd above ten thousand three was 7, and the content of As, Hg and Cu in all the samples was less than 1 ppm, ten thousand one and 20 ppm, respectively. Conclusion:The contents of gallic acid, ellagic acid and total flavonoids can comprehen-sively reflect the content of active ingredients in Melastoma dodecandrum. The contents of acid insoluble ash, Pb and Cd in Melastoma dodecandrum are relative high, which should be studied further to draw up reasonable limit.

Objective To evaluate the effects of vitamin C combined with low dose of gabapentin on neuropathic pain (NP) in rats.Methods Fifty male Sprague-Dawley rats,weighing 225-275 g,aged 6-7 weeks,were randomly assigned into 5 groups (n =10 each):sham operation group (group Sham),NP group,vitamin C group (group VitC),gabapentin group (group Gap),and vitamin C combined with gabapentin group (VitC + Gap group).NP was induced by chronic constrictive injury in the rats anesthetized with pentobarbital sodium.The sciatic nerve was exposed and 4 silk ligatures were placed on the sciatic nerve at 1 mm intervals.In VitC,Gap and VitC+ Gap groups,vitamin C 500 mg/kg,gabapentin 30 mg/kg,and vitamin C 500 mg/kg + gabapentin 30 mg/kg diluted in normal saline 1 ml were injected intraperitoneally,respectively,twice a day for 7 consecutive days starting from 7 days after NP.Normal saline 1 ml was given in Sham and NP groups.Before NP and at 3,7,and 14 days after NP,the paw withdrawal threshold to mechanical stimuli (PWMT) and paw withdrawal latency to thermal stimuli (PWTL) were measured.At 14 days after NP,the blood samples were drawn from the carotid arteries.The rats were then sacrificed and the spinal cord was removed for determination of concentrations of malondialdehyde (MDA) in serum (by using Thiobarbituric acid method) and Slc23a2 mRNA expression (by RTPCR).Results Compared with Sham group,PWMT and PWTL in left paws were significantly decreased in NP,VitC,Gap ard VitC + Gap groups,the concentrations of MDA in serum were increased and Slc23a2 mRNA expression was down-regulated in NP group,Slc23a2 mRNA expression was down-regulated in VitC and Gap groups,and no significant change in Slc23a2 mRNA expression was found in VitC + Gap group.Compared with NP group,PWMT and PWTL in left paws were significantly increased,the concentrations of MDA in serum were decreased,and Slc23a2 mRNA expression was up-regulated in VitC + Gap group,the concentrations of MDA in serum were decreased in VitC group,and no significant changes were found in PWMT and PWTL in left paws in VitC and Gap groups.There were no significant differences in PWMT and PWTL in fight paws at each time point between the five groups.Conclusion Vitamin C combined with low dose of gabapentin can relieve NP effectively,and up-regulation of Slc23a2 mRNA expression,increased trafficking of vitamin C to neurons and inhibition of oxidative stress may be involved in the mechanism.

To study the mechanisms whereby cisplatin suppresses survival of human esophageal squamous cell carcinoma cells.The cytotoxicity of cisplatin in cisplatin-resistant cell line EC109 /CDDP and its parental cell line EC109 was measured by cell viability assay.Western blotting was used to investigate the protein expression of to-tal p53 and phosphorylated p53 at Ser15.Colony formation assay was employed to evaluate the ability of cells to recover from treatments and form colonies.The results indicated that EC109 /CDDP cells were more resistant to cisplatin-induced cytotoxicity than EC109 cells,with the IC50 values of (20.4 ±4.4)μmol /L and (5.7 ±0.1 )μmol /L,respectively.Although cisplatin did not alter the total protein level of p53,it obviously increased the phosphorylation of p53 at Ser15.Cisplatin inhibited survival of both EC109 /CDDP and EC109.Notably,inhibition of p53 by Pifithrin-αsignificantly promoted recovery of cisplatin-treated EC109 and EC109 /CDDP cells to differ-ent degrees.In this respect,p53 protein was found to be activated in response to cisplatin treatment in both EC109 /CDDP and EC109,which may contribute to the cytotoxic effect of cisplatin.

Objective To establish a high performance liquid chromatographic ( HPLC) method for determination of gallic acid and ellagic acid in Melastoma dodecandrum. Methods HPLC was applied, the chromatographic column was Waters Xbidge C18(4.6 mm×250 mm,5μm);with methanol-mixed aqueous solution containing 0.3% phosphoric acid and 0.06%tetrahydrofuran as the mobile phrase;gradient elution:0–12 min,1% A ( the detection wavelength was 216 nm);12–24 min, 1% A→55% A (the detection wavelength was 254 nm);flow rate was 1.0 mL.min-1, the column temperature was 30 ℃. Results Good linear relationship of gallic acid and ellagic acid was obtained at ranges of 0.089 7-0.448 μg ( r=0.999 8) and 0.028 4-0.142 μg (r=0.999 8),respectively. The average recoveries were 96.51% (RSD=1.10%) and 96.19% (RSD=1.20%) . Conclusion The method is simple,accurate and repeatable,and it is suitable for content determination of gallic acid and ellagic acid in Melastoma dodecandrum. It provides a basis for quality evaluation of Melastoma dodecandrum.

Objective To investigate the expression of cyclooxygenase-2(COX-2)and Survivin in ameloblastoma (AB) tissues. Methods A total of 60 AB samples (primary AB 40 cases, recurrent AB 20 cases) and 60 normal oral mucosas were collected from the Affiliated Hospital of Chengde Medical College. The AB samples included 12 acanthomatous types, 18 follicular patterns, 18 plexiform patterns, 6 basal cell ameloblastomas and 6 desmoplastic ameloblastomas. The expres?sion levels of COX-2 and Survivin were detected by immunohistochemistry and Western blot methods in AB and normal oral mucosa. The expressions of COX-2 and Survivin were compared between different types and tissues of AB. Results The positive rate of COX-2 was significantly higher in AB (82.7%) than that in normal oral mucosa (10.0%). The positive rate of Survivin was significantly higher in AB (83.3%) than that in normal oral mucosa tissues (16.7%). The expression of COX-2 (0.781±0.142) was higher in AB than that of normal oral mucosa tissues (0.179±0.056). The expression of Survivin (0.447± 0.139) was significantly higher in AB than those in normal oral mucosa tissues (0.072±0.017). There was a positive correla?tion between the expression of COX-2 and Survivin (r=0.778,P<0.05). There were no significant differences in the positive expressions of COX-2 and Survivin between different types of AB (P>0.05). Conclusion COX-2 and Survivin were over-expressed in AB, which may be involved in the occurrence and development of AB.

Objective To investigate the sodium and potassium disturbances in 9870 elderly inpatients and to analyze their risk factors.Methods Clinical data of sodium and potassium levels and the possible risk factors were collected in the elderly inpatients from a single center.The incidence of sodium and potassium disturbances and their risk factors were analyzed by multiple Logistic regression,and the correction of the imbalance was investigated.Results A total of 6027(61.6％)times of sodium and potassium disturbances were found in the 9870 elderly inpatients on admission and during hospitalization,and the total incidence of this disturbances was 61.6％.The incidences of hyponatremia,hypokalemia,hypernatremia and hyperkalemia were 27.9％ (2729/6027),9.7％ (951/6027),15.4％(1506/6027) and 8.6％ (841/6027),respectively.Heart failure was the common risk factor for the two electrolyte disturbances.T2DM caused hyponatremia,hypokalemia and hypernatremia.Among the medication risk factors,diuretics were the common risk factor for various electrolyte disturbances.Patients taking antiepileptics,antidementia drugs,antidepressants and benzodiazepines were more likely to have hyponatremia.The correction rate of mild,moderate and severe hyponatremia/ hypokalemia were 79.2％(1253/1582),68.1％ (535/786),45.1％ (163/361) and 79.5％(776/976),66.4％(217/327),40.1％(81/203),respectively.The time for the correction of the above degrees of hyponatremia/ hypokalemia were(3.7±2.7) d,(4.1±2.3) d,(8.9±1.6) d and(2.5± 1.4) d,(3.2 ± 1.5) d,(6.1 ± 1.2) d.The supplement amounts of sodium chloride and potassium chloride were(5.98±3.67) g,(9.45±3.02) g,(10.26±1.32) g and(2.23±0.93) g,(5.12± 1.53) g,(8.07 ± 2.46) g,respectively.Conclusions The incidences of electrolyte disturbances are high in elderly inpatients with combined diseases and application of various drugs,and the correction of disturbances is difficult.More attention should be paid to the electrolyte disturbances,which should be corrected positively.

Human Interleukin-11 (hIL-11) is a multifunctional cytokine which plays an important role in regulating the proliferation and differentiation of cells in the hematopoeitic,lymphoid system etc. To obtain the IL-1 1 cDNA, a primary culture of Chinese fetal lung fibroblast was prepared from fresh tissue. Then the human IL-11 cDNA without the N-terminal signal peptide sequence was cloned by RT-PCR from the cells induced by PMA. The sequnce indicated that there are three bases different from those previously reported,but with no change of the amino acids. The cDNA was inserted into the 3′ end of trxcA gene in thioredoxin gene fusion expression system pTRXFUS to construct the trxA and hIL-1 1 fusion expression vector, and expressed in E. coli. The fusion hIL-11 accounts for more than 20 % of the total bacteria proteins.The expression product is present in soluble forms and has the full biological and immunological activities.

To evaluate the regulating effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression in hippocampus tissues of rats with cerebral ischemia, SD rats were divided into the sham-operation group, the model group, the Buyang Huanwu decoction group (ig, 3.15 g · kg⁻¹) and the simple prescription group (ig, 2.41 g · kg⁻¹). Each group was further divided into five subgroups based on time points after the administration, i. e. 1 d, 3 d, 7 d, 14 d and 28 d, respectively. CDK4/Cyclin D1 expressions of the group at different time points were examined by using immunohistochemistry and real-time qPCR. According to the results, the cerebral ischemia model group showed higher CDK4/Cyclin D1 expression than the sham-operation groups (P < 0.05), suggesting that the cell cycle signal pathway would be activated by the cerebral ischemic injury. Both Buyang Huanwu decoction and simple prescription groups showed significantly lower cyclin expression than the model group at 3 d, 7 d, 14 d, 28 d (P < 0.05), indicating both Buyang Huanwu decoction and its simple prescription could play the neuroprotective effect through the cell cycle signal pathway.
Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Cycle ; drug effects ; Cyclin D1 ; genetics ; metabolism ; Cyclin-Dependent Kinase 4 ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley