Objective:To establish a sensitive and fast HPLC method for determining cucurbitacin IIa in rat plasma and investigate the pharmacokinetic characteristics after intravenous administration. Methods: The analysis was performed on a Dikma Diamonsil C18 column (250 mm × 4. 6 mm, 5 μm) at 35℃ with the mobile phase of acetonitrile-water (45∶ 55, v/v) and the UV detection at 212 nm. The plasma samples were collected after intravenous administration at different time points and measured by HPLC. The pharmaco-kinetic parameters were calculated by DAS 2. 0 software. Results: The calibration plot of cucurbitacin IIa was linear over the range 0. 146-14. 060 μg · ml-1 . The range of method recovery was 99. 02%-104. 22% and that of extraction recovery was 84. 74%-86. 80%. The intra-and inter-day precision were both less than 5%. The stability met the requirements. The main pharmacokinetic pa-rameters were showed as follows:t1/2β(h):0.732 ±0.151, 0.681 ±0.055,0.667 ±0.064;Vd (L·kg-1):0.147 ±0.089, 0.131 ±0.095,0.153±0.047; Cl(L·h-1·kg-1): 0.287±0.031,0.304±0.063,0.318±0.029andAUC0→∞(mg·h·L-1):3. 646 ± 1. 124,4. 916 ± 1. 227,9. 385 ± 1. 419. Conclusion: The validated method is successfully applied in basic pharmacokinetic study in rat plasma after intravenous administration. The plasma concentration-time curves at three dosages are all fitted three-compart-ment model. Within the examined dose range, the pharmacokinetics of cucurbitacin IIa in rat is based on linear pharmacokinetics.

Obiective To investigate the effect ofisoflurane on expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA in the hippocampus of immature rats.Methods sixty-four 7-clay-old SD rats were randomly assigned into 2 groups(n=32 each):control group(group C)and isoflurane group(group S).group S was exposed to 1.5% isoflurane for 6 h while group C to air.Fore animals were killed before anesthesia(T0,baseline),at 2,4,6 h(T1-3)of isoflurane anesthesia and 4,6,12 and 24 h after anesthesia(T4-7).The hippocampi were immediately removed for determimation of the expression of IL-1β mRNA,IL-6 mRNA and TNF-α mRNA by RT-PCR.Results Compared with group C,the expression of IL-1β mRNA at T1-5,IL-6 mRNA at T2.3 and TNF-α mRNA at T1-6 in the hippocampus was upregulated in group S.Conclusion The expression of IL-1β mRNA,IL-6 mRNA and TNF-β mRNA was elevated in the hippocampus of immature rats after being exposed to isoflurane.

Objective To investigate the signaling pathways involved in the activation of neuron and glia in spinal cord in rats with neuropathic pain. Methods Twelve female SD rats (weighted 150 to 200 g) were randomized into two groups of spared nerve injury(group SNI) and control(group C). Surgery was performed to build model of SNI neuropathic pain in group SNI. Foot-lift response frequency to mechanical stimulation for ipsilateral hindpaw was assessed by 12 g and 2 g touch stimulator at different times. On the 11~(th) day after operation, 3 rats from each group were fixed by perfusion and the expressions of mitogen-activated/extracellular signal-regulated kinase (MEK), p-mitogen-activated/extracellular signal-regulated kinase (p-MEK), p-extracellular regulated protein kinase(p-ERK) and nuclear factor kappa B (NF-κB) were detected by immunohistochemistry method. And proteins from ipsilateral LA-6 spinal cord in other 3 rats from each group were extracted for Western Blot analysis. Western Blot and immunohistochemistry were performed with antibodies specific for MEK, p-MEK, pERK and NF-κB. Results All rats in group SNI developed a relative unchangeable mechanical allodynia since the 5~(th) day after operation. The results of immunohistochemistry method showed that the expression of MEK was mainly in cytoplasm, p-MEK in cell nuclear, p-ERK in astrocyte and NF-κB in neuron according to morphologic observation. Western Blot analysis indicated that the expressions of p-MEK, p-ERK and NF-κB in group SNI were increased significantly compared with those in group C(P<0. 05). Conclusion In the spinal cord of rats with neuropathic pain, MEK-ERK signaling pathway is activated in astrocytes and NF-κB in neurons, which may contribute to the development of neuropathic pain.

Objective To studied the effects of administering a second rinse solution of low-potassium-dextran(LPD) before reperfusion on the donor lung of rat after long time,cold ischemic preservation.Methods Twenty-four rats were randomly divided into control group(n=12) and trial group(n=12).Donor lungs of control group were flushed with LPD solution.The levels of malonaldehyde(MDA),myeloperoxidase(MPO) and interleukin-8(IL-8) were tested in the tissue of right lung and the left lungs were reperfused with venous blood for 30 minutes.The partial pressure of oxygen(PaO_2),PaP and PawP were measured at every 10 minutes intervals during reperfusion.After reperfusion,the MDA,MPO activity and interlukin-8(IL-8) were determined in the lung tissue.Results The levels of MDA,MPO and IL-8 decreased significantly in right lung of trial group than those in control group(P

Objective To summarize the diagnostic and therapeutic experience on chronic pancreatitis characterized by a pancreatic mass. Methods The clinical data of 28 cases of chronic pancreatitis with mass undergoing surgical operations were retrospectively analyzed in our hospital from June1999 to June 2009. Results Among the 28 cases, 19 were diagnosed as carcinoma, 9 cases were diagnosed as chronic pancreatitis respectively before operation. Needle aspiration biopsy and/or postoperative pathology identified chronic pancreatitis in all cases. The symptom included abdominal pain (22 cases),jaundice (15 cases), and obstruction of duodenum (4 cases). Pancreaticoduodenectomy was performed in 17 cases, choledochojejunostomy performed in 3 cases, pancreatojejunostomy performed in 1 case.Duodenum-preserving resection was performed in 4 cases, and resection of body and tail of the pancreas were performed in 3 cases. There was no operative death. Postoperative complications included pancreatic leakage (2 cases), severe gastroplegia (2 cases) and stress peptic ulcer with massive bleeding ( 1 case). All patients got follow-up ranging from 6 months to 5 years. Recurrence of abdominal pain developed in 7 cases after 2 years. Canceration of pancreatic mass was found respectively in 8 months, 1 year after operation in one each cases. Conclusion Preoperative differential diagnosis of chronic pancreas and pancreatic tumor was difficult. Although needle aspiration biopsy is the effective method for diagnosis, there may be still a possibility of missed diagnosis/misdiagnosis.

Objective:To investigate YKL-40-mediated inflammation in human bronchial epithelial cells and analyzed the soluble factors secreted by bronchial epithelial cells exposed to YKL-40 that were responsible for increasing proliferation and migration of primary normal human bronchial smooth muscle cells (BSMCs).Methods:YKL-40-induced inflammation was assayed in two human bronchial epithelial cells (BEAS-2B cell line and primary human bronchial epithelial cells ,namely HBECs).In addition,we treated BEAS-2B cells and HBECs with YKL-40,and added the conditioned culture media ( YKL-40-BEAS-2B-CM) and ( YKL-40-HBECs-CM) to BSMCs.The proliferation and migration of BSMCs were determined by premixed WST-1 cell proliferation reagent and QCM chemotaxis migration assay ,respectively.Results: Bronchial epithelial cells treated with YKL-40 resulted in a significant increase of IL-8 production,but have no effect about RANTES ,Eotaxin and TNF-α.YKL-40-BEAS-2B-CM and YKL-40-HBECs-CM induced IL-8 was found to further stimulate proliferation and migration of BSMCs ,and the effects were inhibited after neutralizing IL-8.Conclusion:Through investigating the interaction of airway epithelium and smooth muscle ,our findings implicate that YKL-40 may be involved in the inflammation of asthma by induction of IL-8 from epithelium,subsequently contributing to BSMCs proliferation and migration.Moreover, inhibition of IL-8 signaling is a potential therapeutic target for YKL-40-induced inflammation and remodeling of asthma.

Objective To investigate the effects of isoflurane anenthesia on myocyte enhancer factor 2(MEF2) signaling pathway in neonatal rat hippocampus. Methods Twenty-four 5-day-old SD rats of both sexes,weighing 10-13 g, were randomly divided into 2 groups ( n = 12 each): control group (group C) and isoflurane group (group I). In group I, 1.5% isoflurane in 100% O2 was inhaled for 6 h. Group C received no treatment.Three rata in each group were sacrificed at 2, 4, 6 h of isoflurane anenthesia and 24 h after isoflurane anenthesia (T1-4), and the hippocampi removed for determination of MEF2 mRNA, synGAP Ⅰ mRNA, Arc mRNA and synapsinⅠ mRNA expression (by PT-PCR) and synapsin Ⅰ protein expression (by Western blot).Results Compared with group C, the expression of MEF2 mRNA, synGAP Ⅰ mRNA, Arc mRNA and synapsin Ⅰ mRNA at T1-3 and synapsin Ⅰ protein at T2-4 was up-regulated in group I ( P ＜ 0.05). Conclusion Inhalation of anaesthetic concentration of isoflurane may affect synapse formation during the development of central nervous system by actirating hippocampal MEF2 signaling pathways in neonatal rats.

Objective To observe the effect of treating Diabetic Peripheral Neuropathy with Acupoint Injection. Methods 65 patient with DPN were randomly recruited into a control group and a treatment group after adjusting stage. The control group was treated with acupuncture, while the treatment group was treated with the Safflower inoculation fluid acupuncture point injects. Nerve function parameter (MDNS、 NCV) and clinical symptoms were observed after 3 therapeutic courses. Results Nerve function parameter (MDNS、 NCV) and clinical symptoms in the treatment group were apparently improved after treatment (P<0.05) . Conclusion Acupuncture point injected with Safflower inoculation fluid may improve clinical symptoms and nerve functions of DPN patients.

Objective To evaluate the inhibitive effect of tea polyphenol on the growth of human nasopharyngeal carcinoma HONE1 cell xenograft in nude mice ,and to explore the underlying mechanisms .Methods Tumor model was established by subcu‐taneous inoculation of nasopharyngeal carcinoma cell HONE1 into nude mice ,was used to evaluate the antitumor effect of tea poly‐phenol in vivo .The expression levels of VEGF were detected by real‐time PCR and western blot .Results The growth of xenograft in nude mice was significantly suppressed after application of tea polyphenol at a dose‐dependent manner .To compare with control group ,the inhibition rates were 18 .82% (P<0 .05) and 47 .66% (P<0 .05)when treated at low and high dose respectively ,With in‐creased concentration of TP ,the inhibition rates increased .Real‐time fluorescence quantitative‐PCR and western blot results showed that the expression of VEGF decreased at a dose‐dependent manner .The change of high dose group was obviously ,the difference was statistically significant(P<0 .05) .Conclusion Tea polyphenol could significantly inhibit the growth of human nasopharyngeal carcinoma HONE1 cell xenograft in nude mice ,probably by down regulating the VEGF protein level to inhibit tumor angiogenesis effects .

OBJECTIVE: To investigate the effect and mechanism of tea polyphenol (TP) on the proliferation, apoptosis, migration and invasion of nasopharyngeal carcinoma(NPC) cell line HONEl. METHOD: After treated with different concentration of tea polyphenol, CCK-8 assay, fluorescent staining, cell scratching assay and transwell assay were applied to detect the effect of tea polyphenol on the HONE1 cells. Furthermore, the expression of protein VEGF was investigated by flow cytometry assay. RESULT: It was found that tea polyphenol could inhibit NPC cell proliferation significantly in a dose-dependent manner, however, little impact was observed in normal nasopharyngeal epithelial cell line NP69. Furthermore, it was demonstrated by fluorescent staining assay that tea polyphenol could induce NPC cell apoptosis, and cell scratching assay and transwell assay showed that tea polyphenol could inhibit cell migration and invasion. CONCLUSION Tea polyphenol can significantly inhibit cell proliferation, induce cell apoptosis and decreased the migration and invasion ability of NPC cells in vitro. Tea polyphenol might be a tumor suppressor of NPC cells.
Apoptosis ; drug effects ; Carcinoma ; Cell Line, Tumor ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; Polyphenols ; pharmacology ; Tea ; chemistry