Aim To investigate the effect and mechanism of action of chronic morphine treatment on the neurosteroidogensis of primary cultured rat cerebral cortical neurons(CCNs).Methods The effect of morphine on the production of pregnenolone(PREG),dehydroepiandrosterone(DHEA),allopregnanolone(AP),pregnenlolone sulfate(PS)and dehydroepiandrosterone sulfate(DS)in cell culture media was measured by solid-phase extraction and LC-MS,with methyltestosterone(MT)or estrogen sulfate(ES)as internal standards.The dependence-like changes of CCNs were determined by testing the p-CREB level in the nuclear lysates using western blot.Results Compared with the control group,morphine treatment significantly reduced levels of PREG and DS respectively;opioid agonist DAMGO significantly reduced levels of PREG,DS and PS respectively,and increased the level of AP significantly.Compared with the morphine group,?-opioid-antagonist CTAP concomitant with morphine increased the level of PREG significantly.Compared with the control group,chronic morphine treatment or DAMGO treatment significantly increased the level of p-CREB in the CCNs.Compared with the morphine treatment group,?-antagonsit CTAP significantly reduced the level of p-CREB.Conclusion ?-opioid receptor mediated the inhibitory effect of morphine on levels of neurosteroids,and changes of neurosteroid levels might be related to morphine dependence.

Objective To evaluate the changes in the expression of cytochrome cholesterol side chain cleavage enzyme (P450scc) in the brain of morphine-dependent rats. Methods Twenty-four male SD rats aged 4-8 months weighing 180-200 g were randomly divided into 3 groups (n = 8 each): group Ⅰ normal saline (group NS), group Ⅱ morphine dependence (group MD) and group Ⅲ morphine withdrawal (group MW). In group MD and MW, the rats were given intraperitoneally increasing doses of morphine starting from 5 mg/kg to 10, 15, 20, 30, 40 and 50 mg/kg twice a day for 7 days. In group NS, the rats were given equal volume of normal saline instead of morphine. The rats were decapitated 1 h after last injection in group NS and MD. In group MW, naloxone 2 mg/kg was given 1 h after last injection, and then the animals were decapitated 30 min after withdrawal symptoms were observed. The brains were immediately removed and the frontal cortex, hippocampus, striatum and thalamus were separated. The expression of P450see was determined by Western blot. Results The expression of P450scc in the frontal cortex, hippocampus and striatum was significantly decreased in group MD and MW compared with group NS (P<0.05). Conclusion The down-regulation of P450scc expression might be involved in the development of morphine dependence, but it is not involved in the morphine withdrawal.

OBJECTIVE:Alfuzosin concentration in human serum was determined by RP-HPLC METHODS:The mobile phase was methanol-acetonitrile-phosphate buffer-triethylamine(10∶30∶60∶0 05) After being extracted with ethyl ace_tate,alfuzosin was analyzed by reversed-phase HPLC (Shim-pack CLC-ODS,4 6mm?150mm,5?m)and fluorescence detection with excitation wavelength set at 334nm and emission at 378nm RESULTS:Over the concentration range of 0 4～51 2?g/L,the linear regression equation was A=0 3 909C+0 0 605(n=8,r=0 9 996) The average recovery of alfuzosin was 99 4% The intra-and inter-day RSDs were less than 15% CONCLUSION:The method is accurate and can be used for studying the pharmacokinetics of alfuzosin

AIM To investigate if morphine addiction and relapse to morphine-seeking is related to the change in neurosteroid levels in the brain of rats. METHODS Rats were injected ip morphine (5 mg·kg-1·d-1, 18:00-20:00) and trained in conditioned place preference (CPP) box, once daily for 10 d. CPP test was performed 24 h after the last training. After discontinuation of training for 7 d for CPP extinction, then intermittent and inescapable foot-shock (FS, 0.5 mA, 0.5 s on, 40 s off, 15 min) was applied to rats as the priming stimuli for relapse. CPP test was performed 2 h after FS. When CPP test finished, rats were decapitated and the levels of neurosteroids were analyzed using gas chromatography/mass spectrometry. RESULTS CPP was established when rats were injected morphine and trained for 10 d. At the same time, the levels of pregnenolone and allopregnanolone in the brain tissues of rats were significantly increased. When CPP was reinstated in morphine-treated rats by FS-stress after 7 d CPP extinction, the levels of dehydroepiandrosterone and dehydroepiandrosterone sulfate were significantly increased. CONCLUSIONThe development of morphine addiction and relapse may be related to endogenous neurosteroids in rat brain tissues.

A case of benign follicular neoplasm-trichogerminoma-is reported.A 48-year-old man presented with a 10-year history of asymptomatic subcutaneous nodule in the chest.Histological examination revealed a well-circumscribed lesion composed of variously sized lobuli and cysts in the deep dermis and separated from the surrounding tissue by a fibrous capsule.Most lobuli consisted of concentrically arranged clear cells in the central area and basophilic cells in a palisade arrangement in the peripheral area.The tumor cells displayed a multi-directional differentiation toward hair bulb,inner root sheath,outer root sheath and infundibulum of hair follicles.Immunohistochemically,the tumor cells expressed AE1/AE3,CK5/6 and CK17,but were negative for CK20 or CK7.There was a sharp contrast in immunohistochemical findings between the central clear cells and peripheral basophilic cells.Based on the histological and immunohistochemical features,a diagnosis of trichogerminoma was made.

Aim To investigate effects of glutamate(GLU) and ?-aminobutyric acid(GABA) on levels of neurosteroids synthesized and released by primary cultured rat brain cortical astrocytes.Methods Primary cultured rat brain cortical astrocytes were treated with GLU or GABA for 48 h respectively.Unconjugated(dehydroepiandrosterone,DHEA;pregnenolone,PREG;allopregnanolone,AP) and conjugated neurosteroids(dehydroepiandrosterone sulfate,DHEAS;pregnenolone sulfate,PREGS) in culture media were extracted by solid phase extraction(SPE) and analyzed by HPLC-MS.Results Compared with saline control group,GLU treatment decreased PREG and PREGS levels and increased DHEAS level significantly;GABA treatment decreased PREG level and increased AP level significantly.Conclusion Both GLU and GABA were shown to inhibit the genesis of PREG.While GLU significantly promoted the genesis of DHEAS and GABA promoted the genesis of AP.The genesis of PREGS was also inhibited by GLU treatment in primary cultured cortical astrocytes.

Objective To determine the optimum dose of normal saline (NS) for preloading epidural space required to prevent the injury to blood vessel by epidural catheter placement for caesarean section.Methods Two hundred ASA Ⅰ or Ⅱ parturients with a single baby at full term in vertex presentation,aged 24-35 yr,weighing 63-78 kg,scheduled for caesarean section under continuous epidural anesthesia,were randomly divided into4 groups (n-50 each):control group (group Ⅰ),NS2 ml group (group Ⅱ),NS5 ml group (group Ⅲ)and NS 10 ml group (group Ⅳ).The epidural puncture was performed at L2-3 interspace with a Tuohy needle attached to a 5 ml syringe.Loss of resistance was used to identify the epidural space.In group C no fluid was injected into the epidural space before insertion of the catheter,while in groups Ⅱ,Ⅲ and Ⅳ NS 2,5 and 10 ml were injected into the epidural space before the catheter insertion respectively.After a test dose of 3 ml 1.5％ lidocaine,0.75％ ropivacaine 10-20 ml was administered through the epidural catheter.MAP and HR were recorded before epidural puncture (T0),at 10 and 20 min after the end of epidural administration (T1.2),and at the end of surgery (T3).The number of patients in whom blood or blood tinted fluid was withdrawn from the epidural catheter was recorded.The amount of ropivacaine consumed was recorded.The upper level of anesthesia was measured by pin-prick and the degree of motor block was assessed using modified Bromagc scale at T2.Results The hemodynamic parameters were in the normal range in the four groups.MAP was significantly lower at T2,the upper level of anesthesia was significantly higher,and the degree of motor block was significantly smaller in group Ⅳ than in groups Ⅰ,Ⅱ and Ⅲ (P ＜ 0.05).There was no significant difference in MAP among groups Ⅰ,Ⅱ and Ⅲ (P ＞ 0.05).There was no significant difference in HR and the amount of ropivacaine consumed among the four groups (P ＞ 0.05).The number of patients in whom blood or blood tinted fluid was withdrawn fiom epidural catheter was significantly smaller in groups Ⅲ and Ⅳ compared with groups Ⅰ and Ⅱ (P ＜ 0.05).Conclusion Preloading the epidural space with NS 5 ml can prevent the occurrence of injury to blood vessel induced by insertion of epidural catheter with no influence on the efficacy of anesthesia and NS 5 ml is the optimum dose.

ObjectiveTo evaluate the effect of preloading epidural space with 0.9％ sodium chloride on the incidence of the injury to blood vessel by epidural catheter insertion for cesarean section.Methods One hundred uterogestation patients with single birth,ASA class Ⅰ - Ⅱ,underwent caesarean section and requested continuous epidural analgesia were divided into group P and group C with each 50 cases by random digits table.After identification of the epidural space,group C was inserted epidural catheter directly,and 5 ml of 0.9％ sodium chloride was injected into epidural space through the epidural needle in group P,while the syringe plunger was held closed for 20 s to make sure the solution spreaded sufficiently,following insertion the epidural catheter.Between the two groups,mean arterial pressure and heart rate were recorded prior to anesthesia,2 min after turn to the supine horizontal position after succeeded puncture,the time when the fetus were born and when the surgery were over.The cases with bloody fluid in the epidural puncture needle during puncture,or in the epidural catheter during catheter placement,fresh blood in the epidural catheter,and bloody fluid in caudal end of epidural catheter during extubation were recorded.ResultsThe changes of mean arterial pressure and heart rate were all in the normal range,there was no obvious difference between the two groups.The incidence of rates with bloody fluid in the epidural puncture needle during puncture and the bloody fluid in caudal end of epidural catheter during extubation in group P were significantly lower than those in group C [ 10％ (5/50) vs.26％ (13/50),22％ (11/50) vs.48％ (24/50),P ＜ 0.05 or ＜ 0.01 ].ConclusionPreloading epidural space with 5 ml 0.9％ sodium chloride can reduce the incidence of the injury to blood vessel induced by insertion of epidural catheter.

Two cases of atrophic dermatofibrosarcoma protuberans (DFSP) are reported.The patients were a 16-year-old and a 24-year-old boy with a clinical course of 6 and 5 years respectively.The lesions began as well-marginated atrophic erythema,and subcutaneous nodules appeared gradually on the erythema.Histopathology showed atrophic or normal epidermis,wavy arrangement of tumor (spindle) cells in the superficial dermis which was aligned parallel to the epidermis,storiform arrangement of tumor cells in the lower dennis,and typical lacelike pattern of infiltration of subcutaneous fat tissue with tumor cells.Immunohistochemistry showed that the tumor cells stained positive for vimentin and CD34,but negative for S100 or CD 68.

Objective To investigate the relationship between the clinical manifestations,prognosis and histopathological findings of mucormycosis.Methods The clinical data on and pathological findings from 7 cases of mucormycosis confirmed by fungal culture in the institute from 1989 to 2006 were analyzed retrospectively.Results There was 1 case of hinocerebral mucormycosis and 6 cases of cutaneous mucormycosis,among them,2 were mucormycotic necrotizing fasciitis (MNF).The condition of patients with rhinocerebral mucormycosis or MNF aggravated rapidly and all the 3 patients died from mucormycosis. Histopathological examination showed mixed infiltrates of inflammatory cells as well as necrosis and angioin vasion.On the contrary,the condition of the remaining 4 patients with cutanesus mucormycosis,who presented mainly with indurated erythematous patch,progressed slowly,and 2 patients were cured.Histologically,the lesions were characterized by granulbmatous infiltration with a few hyphae;no typical angioinva sion phenomenon was noted.There was no evidence of perineural invasion with hyphae in any of the 7 cases.ConclusionIn patients with mucormycosis,histopathological findings characterized by mixed infiltrates of inflammatory cells,numerous hyphae and typical angioinvasion phenomenon may herald a poor prognosis.