Objective To investigate the expression of mRNAs for cholesterol side chain cleavage enzyme (p450 scc), 17?-hydroxylase / C17-20 lyase (P450 C17) and 3?-hydroxysteroid dehydrogenase (3?-HSD) in frontal cortex, amygdala, hippocampus, striatum and midbrain of morphine dependence rats.Methods Twenty-one male SD rats were randomly divided into 3 groups with 7 animals in each group: (1) control group (group C) ; (2) morphine dependence group (group D) and (3) morphine withdrawal group (group W). In group D and W the animals were given intraperitoneally increasing doses of morphine starting from 5 mg?kg-1 to 10, 15, 20, 30, 40 and 50 mg?kg-1 twice a day for 7 days. In group C the animals were given normal saline instead of morphine. In group C and D the animals were decapitated 1 h after last injection. In group W naloxone 2 mg?kg-1 was given 1h after last morphine injection, then the animals were decapitated 30 min after withdrawal symptoms were observed. The brains were immediately removed and the frontal cortex, amygdala, hippocampus, striatum and midbrain were separated. The expression of mRNAs for the three steroidogenic enzymes in the different brain regions of rats were determined by RT-PCR.Results The expression of P450scc mRNA in striatum and 3?-HSD mRNA in amygdala, striatum and frontal cortex decreased in group D compared with group C. The expression of 3?-HSD mRNA increased in morphine withdrawal group compared with group D.Conclusion The gene expression of steroidogenic enzymes decreases in some brain regions of morphine dependence rats, suggesting that endogenous neurosteroids might be involved in 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.

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.

BACKGROUND:Previous studies have demonstrated that normal rat liver homogenate supernatant can induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels with partial hepatocyte functions. However, whether fibrotic liver homogenate supernatant can work or how the inducing effect is remains unclear. OBJECTIVE:To investigate the differentiation potential of human umbilical cord mesenchymal stem cels into hepatocytes under the normal liver and fibrotic liver microenvironment in vitro. METHODS:Liver fibrosis was induced in the SD rats by repeated intraperitoneal injections of 3% thioacetamide at a dose of 200 mg/kg body mass, twice a week for 4 weeks, and then fibrotic liver tissues and normal liver tissues were used to prepare liver homogenate supernatants. Passage 3 human umbilical cord mesenchymal stem cels were used and divided into standard control group (cels were cultured in DMEM/F12 with 10% fetal bovine serum), fibrotic liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 50 g/L fibrotic liver homogenate supernatants), normal liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 100 g/L normal liver homogenate supernatants). The morphological changes of the cels in each group were recorded under inverted microscope; the protein levels of CK18, AFP, CYP3A4, CYP2E1, CYP2D6 and TPH2 were evaluated using western blot assay. Furthermore, the concentration of albumin in the cels was measured. RESULTS AND CONCLUSION:After a 7-day inducement, the stem cels in liver homogenate supernatants groups lost their fusiform shape and changed into hepatocyte-like cels with the morphous of round shape. Compared with the standard control group, the hepatocyte-like cels in the two liver homogenate supernatants groups exhibited human hepatocyte biomarkers, CK18 and AFP. The standard control group cels could express a little amount of CYP2E1, while cels in the two liver homogenate supernatants groups could express CYP3A4, CYP2E1, CYP2D6, TPH2. Compared with the standard control group, the expression level of CYP2E1 in the two liver homogenate supernatants groups increased significantly (P < 0.01), and however, the relative levels of CYP3A4, CYP2E1, CYP2D6, TPH2 in the two liver homogenate supernatants groups showed no statistical significance (P > 0.05). At the same time, compared with the standard control group, the concentration of albumin in the two liver homogenate supernatants groups markedly increased (P < 0.01), but there was no difference between the two liver homogenate supernatants groups (P > 0.05). Experimental findings demonstrated that both of normal liver tissue and fibrotic liver tissue microenvironments could induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels. To achieve the same effect, compared with normal liver tissue, fibrotic liver tissue required lower concentrations, suggesting that fibrotic liver tissue microenvironment may be more conducive to differentiation of umbilical cord mesenchymal stem cels into hepatocytes.

ObjectiveTo investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis in PC12 cells undergone ischemia and hypoxia, and the mechanism involved.Methods PC12 cells at logarithmic phase were collected, and were divided into recombination lentivirus infection group (infected by lentivirus containing HSP70 and fluorescent gene), lentivirus control group (infected by lentivirus containing fluorescin without HSP70 gene) and non-infection group. HSP70 gene and protein expressions in PC12 cells were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. After being challenged with ischemia and hypoxia for 4 hours, the viability of cells was detected by methyl thiazolyl tetrazolium (MTT), the levels of lactic acid dehydrogenase (LDH) in cell supernatant were determined by LDH measurement test kit. The concentration of intracelluer calcium ([Ca2+]i) was assayed by flow cytometer. The activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and total ATPase were measured by ATPase test kits.Results The expressions of exogenous HSP70 gene and protein were found by RT-PCR and Western Blot in the recombination lentivirus infection group. After being challenged with ischemia and hypoxia, the viability of cells in the recombination lentivirus infection group was increased significantly as compared with the lentivirus control group and non-infection group (A value: 0.575±0.020 vs. 0.395±0.014, 0.363±0.045,t1= 17.996,t2= 10.600, bothP< 0.001), the levels of LDH in culture medium and the concentration of [Ca2+]i were decreased significantly [LDH (U/L): 743.46±23.68 vs. 935.43±34.77, 962.89±26.68,t1= 11.179, t2= 15.044, bothP< 0.001; [Ca2+]i relative fluorescence: (31.60±2.43)% vs. (41.48±3.33)%, (40.40±3.05)%, t1= 5.853,t2= 5.502, bothP< 0.001], and the activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and total ATPase were increased significantly [Na+-K+-ATPase (μmol·mg-1·h-1): 8.608±0.307 vs. 6.728±0.173, 6.450±0.091,t1=9.237,P1= 0.001,t2= 11.675,P2< 0.001; Ca2+-Mg2+-ATPase (μmol·mg-1·h-1): 10.523±0.036 vs. 7.910±0.209, 8.064±0.195,t1= 9.718,P1= 0.001,t2= 11.535,P2<0.001; total ATPase (μmol·mg-1·h-1): 17.041±0.324 vs. 14.150±0.182, 13.983±0.085,t1= 16.113,t2= 17.602, bothP<0.001]. There was no statistical difference in above indexes between lentivirus control group and non-infection group.Conclusion HSP70 can maintain the PC12 cells calcium homeostasis, which may be one of the important mechanisms of anti-apoptosis.

Objective To investigate the effect of manual operation on occupational therapy teaching. Methods 83 students were randomly divided into 2 groups: 41 students of general teaching group and the other 42 of manual operation teaching group, who received the ordinary classroom teaching and manual operation teaching respectively. After 4 h of study, they were measured with Canadian Model of Occupational Performance (CMOP). Results There were 37 and 41 questionnaires recalled from the general teaching group and the manual operation teaching group, and 2 and 3 questionnaires excluded as incomplete respectively. The scores of the classroom atmosphere, interesting stimulation, explanation of curriculum connotation, obtaining professional ideology, ability cultivated to solve practical problems, good relationship between doctors and patients developing, and thinking of discussion stimulating were more in the manual operation teaching group than in the general teaching group (P<0.01). Conclusion Manual operation is a new, effective, convenient, and low-cost mode of teaching for occupational therapy.

Objective To study the effects of ankle-foot retractor on ankle joint motion and equilibrium function of stroke hemiplegic patients.Methods 74 patients were divided into two groups: observation group (n=36) and control group (n=38). The control group was trainedby routine rehabilitation training program and electro-uprise bed, the observation group was trained by ankle-foot retractor based on routinerehabilitation training program. The effect was evaluated after 8 weeks. Results After training, either the observation group or the controlgroup showed significant improvement at motion of ankle joint, activity of daily living (ADL), and equilibrium function, but the observationgroup was better than the control group (P<0.001). Conclusion Ankle-foot retractor plays a positive role on motion of ankle joint, ADL andequilibrium function in stroke patients.

Objective To study the effect of motor imagery on balance rehabilitation in hemiplegics after stroke.Methods 20 hemiplegic patients who were in chronic stage(above 6 months)were randomly divided into two groups.Normal rehabilitation was performed in the whole stage and motor imagery was added in group A in stage Ⅰ and group B in stage Ⅱ.All patients were assessed with Berg Balance Scale(BBS),blance subscale of the Fugl-Meyer Assessment(FM-B),max affected limb-loading per weight and Functional Independence Measure(FIM)at the beginning and the ending of the every stage,respectively.Results The deference scores of BBS and FIM,as well as max affected limb-loading per weight between in the stage with motor imagery was better than those in the stage without motor imagery(P<0.01),as well as that of FM-B score(P<0.05).Conclusion Motor imagery can improve balance rehabilitation and activity of daily living in chronic hemiplegics.

Objective To investigate the effect of transcranial direct current stimulation (tDCS) on aphasia recovery after stroke. Meth-ods From April, 2012 to January, 2013, 20 aphasic patients after stroke were enrolled in an A-B experiment design. During phase A, ten times of sham tDCS and language training (five days a week) were implemented, then ten times language training combined with tDCS (five days a week) were implemented in phase B. The treatment lasted for four weeks. Picture naming was measured for all patients before and af-ter treatment both in phase A and phase B. Results The D-value scores of picture naming before and after treatment were significantly more in phase B than in phase A in both treatment items and non-treatment items (t>3.030, P<0.05). Conclusion tDCS could raise the accuracy of picture naming in patients with aphasia after stroke.

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.