The corneal epithelial cells are at the surface of the cornea.Human corneal epithelial cells cultured in vitro contribute to fully understand the biological properties of corneal cells,construct disease models in vitro,and play its clinical significance better.This article reviews the recent advances in human corneal epithelial cells cultured in vitro about extractive fraction,extraction methods,factors of progress,impact identification methods.

OBJECTIVE:To study conditions of disfunction of blood clotting induced by cefoperazoned/sulbactam.METHODS:Cases of disfunction of blood clotting induced by cefoperazoned/sulbactam in our hospital from Mar to May in2004were analyzed statistically.RESULTS:The chief clinical manifestation was alimentary tract hemorrhage,which may relate to the dosage and course of treatment of cefoperazoned/sulbactam.CONCLUSION:The application of cefoperazoned/sulbactam in senile patients should be more prudent.

OBJECTIVE:To provide into the regularity of anaphylactic shock induced by fosfomycin injection.METHODS:The reports of fosfomycin-induced anaphylactic shock from 1994 to 2004 were retrieved and analyzed.RESULTS:36 cases of anaphylactic shock were publicly reported in these 10 years,among which there were 3 cases of death.CONCLUSION:Fosfomycin may induce anaphylactic shock,which should arouse more attention in clinic.

Objective To investigate the effects of morphine dependence and withdrawal on neurosteroids and amino acid transmitters of rat amygdala. Methods Morphine dependence was induced by pretreatment with increasing doses of morphine for 7 days. Withdrawal was precipitated by naloxone (2mg/kg). Withdrawal syndromes were observed and scored. After decapitation, amygdala was dissected out. Nomadic and conjugated neurosteroids were extracted using liquid-liquid extraction and solid phase extraction. Concentrations of neurosteroids including dehydroepiandrosterone (DHEA), pregnenolone (PREG), allopregnanolone (AP), dehydroepiandrosterone sulfate (DHEAS) and pregnenolone sulfate (PREGS) were detected with HPLC-MS. Concentrations of glycine (GLY), glutamate (GLU) and gamma-aminobutyric acid (GABA) were quantitated by HPLC-ECD with pre-column OPA derivatization. Results Compared with saline control, the DHEA level in rat amygdala of morphine dependent group decreased by 33% (P＜0.01). Compared with naloxone control, the PREG and AP levels in rat amygdala of morphine withdrawal group increased by 45% (P＜0.05) and 42% (P＜0.05) respectively; the GABA level decreased by 18% (P＜0.01). Compared with morphine dependent group, the PREG and PREGS levels in rat amygdala of morphine withdrawal group increased by 60% and 40% respectively (P＜0.05); the glycine level decreased by 14% (P＜0.05). Conclusion The DHEA in rat amygdala may play a role in the development of morphine dependence but not involved in the manifestation of withdrawal symptoms. Other neurosteroids (including PREG, AP and PREGS) in rat amygdala seem to be involved in withdrawal but not in dependence. The synthesis and release of inhibitory amino acids in amygdala were depressed when withdrawal was precipitated by naloxone. The results suggest that different changes of neurosteroids and amino acids exist in stages of morphine dependence and withdrawal.

Aim To detect the effect of morphine dependence and withdrawal on the levels of neurosteroids and amino acid neurotransmitters in nucleus accumbens in rat morphine dependent model. Methods Nucleus accumbens was dissected out from morphine dependent and naloxone precipitated withdrawal rats. The contents of neurosteroids including dehydroepiandrosterone, pregnenolone, allopregnanolone, dehydroepiandrosterone sulfate and pregnenolone sulfate were detected with liquid chromatography-negative atmospheric pressure with ionization mass spectrometry(LC-MS). The contents of glycine, glutamate and ?-aminobutyric acid were quantitated by HPLC-ECD with precolumn derivatization. Results Compared with saline group,in nucleus accumbens of morphine withdrawal rats, the level of dehydroepiandrosterone sulfate (P

Objective: To investigate the effects of preconditioning with lithium on the expressions of HSP70 in gerbil after global ischemia. Methods: The transient ischemia model of gerbil global was established by clamping the bilateral common carotid arteries for 5 min.36 gerbils were randomly divided into group lithium(n=18) and group control(n=18).Gerbils in group lithium were injected with lithium intraperitoneally,5 mmol/kg,once a day for 5 consecutive days.Normal saline was used instead of lithium in group control as vehicle control.Both group lithium and control underwent ischemia after preconditioning,and were then further divided into three subgroups(n=6,in each subgroup).The garbles in lithium 1 or control 1 subgroup were sacrificed 1 day after ischemi;lithium 3 or control 3 sacrificed in 3 days;and lithium 7 or control 7 were sacrificed in 7 days.The brains were made into paraffin sections and were used for immunohistochemistry against HSP70. Results: Compared with the vehicle control,the HSP70 expression in subgroup lithium1 was inhibited in the hippocampus CA1(P

Objective: To investigate the effect of morphine dependence and withdrawal on the levels of neurosteroids in hippocampus of male rat.Methods: Rats were given (ip) increasing doses of morphine to form morphine physical dependence, withdrawal syndromes were precipitated by naloxone. The conditioned place preference (CPP) was used to establish morphine psychological dependence. The concentrations of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), pregnenolone (PREG), pregnenolone sulfate (PREGS), and allopregnanolone (AP) in rat hippocampus and plasma were quantified by liquid chromatography-mass spectrometry. Results:The rat model of morphine physical and psychological dependence were successfully established by ip increasing doses of morphine for 7 days and 5mg?kg~ -1 morphine for 10 days respectively. Compared with saline control group, morphine physical dependence increased DHEA and PREG contents in rat hippocampus (0.88?0.19/0.67?0.17,t=2.52,10.94?2.02/7.53?2.64,t=3.24,P

Objective To study the changes in the polyol pathway of glucose metabolism and oxidative stress responses induced by general thoracic surgery by measuring erythrocyte aldose reductase (AR), phosphofructokinase (PFK) and glucose-6 phospate dehydrogenase (G-6PD) activities and plasma MDA and GSH levels before and after surgery performed under different anesthetic techniques. Methods Thirty-two ASA Ⅰ-Ⅱ patients aged 45-71 yrs weighing 55-70 kg were randomly allocated to one of two groups with 16 patients in each group : group Ⅰ received isoflurane inhalation for maintenance of anesthesia and group Ⅱ isoflurane inhalation combined with epidural block. Patients with endocrine and glucose metabolism-related diseases were excluded. The patients were premedicated with intramuscular phenobarbital 0.1 g and atropine 0.5 mg. Anesthesia was induced with intravenous midazolam 0.1 mg?kg-1 , propofol 1.0-1.5 mg?kg-1 , fentanyl 0.1-0.15 mg, droperidol 1-2 mg and vecuronium 0.1 mg?kg-1 in both groups. The patients were mechanically ventilated (VT 8-10 ml?kg-1, RR 10-12 bpm) after tracheal intubation. Anesthesia was maintained with isoflurane inhalation at 1.2-1.6 MAC in group Ⅰ or isoflurane inhalation (0.6-0.8 MAC) combined with epidural block (T9-10) with 1 % lidocaine in group Ⅱ supplemented with fentanyl 5-10 ?g?kg-1 and droperidol 0.1-0.2 mg?kg-1 in divided doses and vecuronium infusion at 1-2 ?g? kg?min-1 . Venous blood samples were obtained before anesthesia (baseline, T0), 90 min after skin incision (T1), 60 min after surgery (T2 ) and on the 1 st and 2nd postoperative days (T3 , T4 ) for determination of erythrocyte AR, PFK and G-6PD activities and plasma glucose, MDA and GSH levels. Results In group Ⅰ plasma MDA level and AR and G-6PD activities in erythrocyte were significantly increased, while plasma GSH level and PFK activity in erythrocyte were significantly decreased on the 1st postoperative day compared to the baseline values before anesthesia (T0) (P 0.05). Conclusion Isoflurane inhalation combined with epidural block can effectively attenuate the effects of surgical trauma on glucose metabolism in theerythrocyte and the oxidative stress responses of the body.

Objective To investigate and compare the effects of pretreatment with different concentrations of trarnadol and morphine on the differentiation of hunan helper T cells (Th) in vitro. Methods Fifteen ASA Ⅰ patients of both sexes (8 males, 7 females) aged 23-59 yrs scheduled for minor day-care surgery were enrolled in this study. The patients had never taken morphine and tramadol before. Blood samples were taken from peripheral vein. Whole blood and mononuclear cells isolated from peripheral blood (PBMCs) were incubated with morphine 10, 100 or 1000 ng?ml-1(group M1 , M2 , M3) or tramadol 50, 500, 5000 ng?ml-1(group Q1, Q2, Q3) for 24 h. Then irrntants was added and incubated for another 5 or 24 h. In the whole blood the level of IL-2 and IFN-?(as markers of Th1 cells) and IL-4 and IL-10 (as markers of Th2 cells) were determined using three-color flow cytometry. In PBMCs the expression of CD4+ and CCR5+ (as markers of Thl cells) and CD4+ and CCR3+ (as markers of Th2 cells) were determined. The Th1/Th2 ratio was calculated. Results Pretreatment with morphine or tramadol significantly increased the number of Th2 cells while the number of Thl cells significantly decreased both in a dose-dependent manner leading to a significant decrease in Th1/Th2 ratio. Conclusion Tramadol and morphine can inhabit in vitro the cellular immune function in dose-dependent manner. The immunosuppressive effect of tramadol is less than that of morphine.

Objective:To establish the quality standard for infantile bronchitis traditional Chinese medicine lotion. Methods:TLC was used for the qualitative identification of Ephedra herba, Paeonia lactiflora Pall. , Poria cocos, Fructus aurantii and Epimedium brev-icornu Maxim. . HPLC was used for the content determination of ephedrine hydrochloride, pseudoephedrine hydrochloride and paeoni-florin. The chromatography conditions for ephedrine hydrochloride and pseudoephedrine hydrochloride were as follows: an AElichrom Pdar-Phenyl Polyphenyl ether-bonded phenyl-bonded silica column (250 mm × 4. 6 mm, 5μm) was used, the mobile phase was aceto-nitrile-0. 2% phosphoric acid solution (1 :99), the flow rate was 1. 0ml·min-1, the detection wavelength was 210 nm, the column temperature was 30℃ and the injection volume was 10 μl. The chromatography conditions for paeoniflorin were as follows:an Inertsil ODS-3 C18 column (250 mm × 4. 6 mm, 5 μm) was used, the mobile phase was acetonitrile-0. 1% phosphoric acid solution (15:85), the flow rate was 1. 0 ml·min-1, the detection wavelength was 230 nm, the column temperature was 30℃ and the injection vol-ume was 10 μl. Results:The TLC results of Ephedra herba, Paeonia lactiflora Pall. , Poria cocos, Fructus aurantii and Epimedium brevicornu Maxim. showed clear spots with good resolution. Ephedrine hydrochloride, pseudoephedrine hydrochloride and paeoniflorin had a good linear relationship within the range of 10.11-101.10 μg·ml-1(r=0.9996), 10.08-100.80 μg·ml-1(r=0.9991) and 20.50-102.5 μg·ml-1(r =0.9996), respectively. The average recovery was 98.80% (RSD =1.87%, n =6), 98.77%(RSD=1. 72%, n=6) and 99. 57% (RSD=1. 56%, n=6), respectively. Conclusion: The established quality standard can be used for the quality control of infantile bronchitis traditional Chinese medicine lotion.