Objective Beta platelet-derived growth factor receptor ( PDGFR-β)-mediated signaling plays a key role in mor-phine tolerance , but its molecular mechanisms are not yet completely understood .The present study aims to investigate whether the ex-tracellular signal-regulated kinase ( ERK) and cyclic AMP response element binding protein ( REB) signaling pathways are involved in the development of PDGFR-βactivation-induced morphine tolerance in rats . Methods Thirty-six adult male SD rats were randomly divided into six groups of equal number:normal saline (20μL), morphine (15μg), morphine +imatinib (morphine 15μg +ima-tinib 10μg), morphine +PDGF-BB (morphine 15μg +PDGF-BB 10 ng), imatinib (10μg), and PDGF-BB (10 ng), all treated intrathecally at 20μL once daily for 7 consecutive days .Paw withdrawal latency ( PWL ) was measured 1 d before and 30 min after medication at 1, 3, 5, and 7 days, respectively, followed by calculation of the maximal possible effect of analgesia (MPE).On the 8th day, PWL was again obtained from all the rats at 30 min after intrathecal injection of morphine (15μg).Then, all the animals were sacrificed and the L4-5 segment of the spinal cord was isolated for determination of the expressions of ERK , phosphorylated ERK ( p-ERK) , CREB, and phosphorylated CREB ( p-CREB) by Western blot. Results At 5 and 7 days after medication, MPE was significant decreased in the morphine group ([52.90 ±8.20] and [15.12 ±3.80] %) and the morphine +PDGF-BB group ([43.51 ±5.42] and [14.81 ±3.60] %) as compared with (100.00 ± 0.00) %in both groups at 1 day (P<0.05), but had no significant changes in the morphine +imatinib group at 1, 3, 5, and 7 days.After intrathecal injection of morphine on the 8th day, MPE was (16.22 ±2.51) %in the morphine group, (15.22 ±3.50) %in the morphine +PDGF-BB group, and (35.21 ±4.51) %in the PDGF-BB group, all remarkably lower than (100.00 ±0.00) %in the control group (P<0.05).There were no significant differences in the expression levels of ERK and CREB among the six groups.The expressions of spinal p-ERK and p-CREB were markedly increased in the morphine , morphine +PDGF-BB, and PDGF-BB groups as compared with the control group (P<0.05), but significantly decreased in the morphine +imatinib group in compari-son with the morphine group, (P<0.05). Conclusion The PDGFR-βsignaling pathway plays an important role in the develop-ment of tolerance to morphine-induced analgesia and its underlying mechanisms may be associated with the activation of the ERK and CREB pathways .

Objective To investigate the expression patterns of fibroblast growth factor 23 (FGF23) in osteoblast and its responses to calcium, phosphate, exogenous PTH and 1,25(OH)_2D~3. Methods The primary rat calvarial osteoblasts were cultured in MEM medium which containing 10% FBS, then were harvested when cells were in half-confluence, confluence, osteoid deposition and osteoid mineralization stages respectively. The procedure was monitored under microscopy. Total RNA was extracted from cells according to the Trizol procedure. FGF23 mRNA levels were determined by Real-time PCR. Further, the confluent osteoblasts were treated with 3.2 mmol/L CaCl_2, 4.4 mmol/L β-glycerophosphate, 10~(-9) mol/L rhPTH(1-34) and 10~(-8) mol/L 1,25(OH)_2D_3 respectively for 3 days, and same volume of the medium was added as the control. The gene expressions were determined by Real-time PCR. Results FGF23 expression was transiently up-regulated at cell confluent stage and down-regulated after that. The FGF23 mRNA levels were 7.5-fold higher in confluent cells compared with that in half-confluent cells (P<0.001). The markedlly stimulating effect (about 16 times) on FGF23 expression was stimulated by exogenous 1,25(OH)_2D_3 treatment while no significant effect was found on FGF23 mRNA levels by CaCl_2,β-glycerophosphate, and rhPTH(1-34) treatments when compared with the control. Conclusions The FGF23 expression in osteoblast is developmental stage-related and its powerful stimulator is 1,25(OH)_2D.

Objective To study causes of deterioration of sudden mydriasis in craniocerebral trauma patients with normal intracranial pressure and verify the efficiency of specific treatments.Methods A retrospective analysis was performed on causes of four cases of accidental mydriasis in normal intracranial pressure among 473 cases of craniocerebral trauma treated from June 2008 to March 2012.Changes of patients' condition and monitoring indices were observed after specific treatments.Results Abnormal mydriasis with synchronously normal intracranial pressure was largely due to sufficient decompression after a certain period of intracranial hypertension and persistence of brain perfusion pressure to more than 110 mm Hg or due to high cerebral perfusion pressure caused by redundant drainage of cerebrospinal fluid or low intracranial pressure (＜ 10 mm Hg),together with factors like low plasma osmotic pressure and carbon dioxide accumulation.The study showed that the intracranial pressure was maintained normal,that the brain swelling took a turn for better,that medical condition were stabilized and that pupil returned to normal in the four cases after treatment with specific protocol.GOS was four points in three cases and five points in one during follow-up at six months postoperatively.Conclusion Incidence of mydriasis with normal intracranial pressure in craniocerebral trauma patients can be efficiently declined through reduction of peripheral blood pressure,perfusion pressure controlling,hypertonic remedy maintenance and brain edema relief.

Obejective To study the effect of daidzein(Daid) on the enzymes of bone metabolism and bone rebuilding in ovariectomized rats.Method Sixty 3-mon female SD rats were divided into 6 groups:Sham,OVX,OVX-D1～D4 groups.Every day the rats in Sham and OVX groups were given distilled water,and the rats in OVX-D1,OVX-D2,OVX-D3,OVX-D4 groups were given 25,50,75,100 mg/kg bw Daid respectively.After 6 mon,all of rats were killed,and the blood,liver and femur were collected.The content of calcium and phosphorus in serum and the enzymes of bone metabolism were measured.Results The content of calcium,phosphorus and ATPase in serum was increased,but the activity of serum alkaline phosphatase(ALP) and tartrate-resistant acid phosphatase(TRAP-5b) was decreased dose-dependently in OVX-D groups compared with OVX group.Conclusion The expression of bone metabolism-associated enzymes could be inhibited,which resulted in inhibition of bone formation and absorbsion in ovariectomized rats.The new balance between bone formation and absorption could be rebuilt by Daid,through the mechanism of decreasing rate of bone conversion.

OBJECTIVE To investigate the pathogenic causes of lower respiratory tract infections(LRTIs) in the elderly in Guangzhou area.METHODS Pathogens obtained from 107 patients with LRTIs were performed by multiple diagnostic tools that including bacterial culture,PCR and specific immunological assays.RESULTS A bacterial cause was established in 42(68.5%) and an atypical pathogen cause in 25(31.6%) of the 107 patients.Mycoplasma pneumoniae and Haemophilus influenzae remained the most important pathogens for LRTIs.CONCLUSIONS In the prescription of antibiotics in the elderly with LRTIs,not only bacteria but also atypical pathogens should be taken into account.

Objective To develop a method of real-time quantitative polymerase chain reaction (RQ-PCR) for detection of translocation ETS leukemia-acute myeloid leukemia 1(TEL-AML1) fusion gene in children with acute lymphoblast leukemia(ALL),and explore its clinical application in minimal residual disease (MRD) monitoring.Methods By reverse transcription and RQ-PCR,a quantifying of TEL-AML1 fusion gene was developed,and the expression levels of TEL-AML1 were detected in bone marrow (BM) samples obtained from 24 children with ALL at diagnosis and at the end of induction of remission,as well as at a series of follow-up. Moreover,the results of MRD detection by RQ-PCR were compared with that of detection by routine morphological examine of BM and cells differentiation mark analysis by flow cytometer(FCM),to evaluate the sensitivity of RQ-PCR in MRD monitoring. Results A method of RQ-PCR targeted at TEL-AML1 fusion gene was established. In 11 BM samples,which collected from TEL-AML1 positive children at the end of induction therapy and all of them achieved completely remission (CR) by routine morphological examine,5 samples were found to be MRD positive by RQ-PCR,positive ratio was 45%. There were 15 BM samples collected in maintenance therapy period,and all these samples were CR by routine morphological examine. However,by RQ-PCR,3 out of 15 samples were found to be MRD positive during maintenance therapy period. After intense and maintenance therapy,the MRD levels of the 3 children were declined to negative. In a recrudescent child,the expression of TEL-AML1 fusion gene was rose by a magnitude of 103 copies before relapse,and after induction therapy once again the patient was completely relaxed.Conclusions RQ-PCR targeted at TEL-AML1 has a higher sensitivity than conventional morphologic way and FCM. RQ-PCR can be used in the quantitative detection of MRD,and provide gist for early prognosticating a relapse and instructing clinical therapy.

ObjectiveTo develop a stable model of focal cerebral infarction in rat to study the curative effect of neural stem cells transplantation.MethodsThirty-seven rats were selected which were divided into two groups in random, experimental group and control group. The focal infarction model was developed by the ligation of the left middle cerebral artery followed by the ligation of the ipsilateral common carotid artery and the temporary clip occlusion of the contralateral common carotid artery for 1.5 h. The operation adopted minimally invasive craniotomy though temporal bone. The model was evaluated by examining the neurologic deficits, ink perfusion, TTC staining and Magnetic Resonance imaging.ResultsAll the rats were in good condition after the operation, the mortality rate was 6.25% after 4 weeks. Ink perfusion and TTC staining confirmed that the ischemia was confined to the cortex. The areas of infarction measured 83.52 mm3 by Magnetic Resonance imaging after 4 weeks.ConclusionA stable focal cerebral infarction model can be achieved by minimally invasive craniotomy. It is superior for its homogeneity of infarction volume and site, and its low mortality. It can be used for the study of transplantation of neural stem cells.

Objective To improve treatment success rate and prognosis for patients with bifrontal contusions by intracranial pressure monitoring.Methods A retrospective analysis was conducted on 79 cases of bifrontal contusions admitted between October 2004 and April 2012.The patients were divided into intracranial pressure monitoring group (n =40) and group without intracranial pressure monitoring (n =39),according to the treatments.Significance of high coronary craniotomy timing,surgical strategy and intracranial pressure monitoring in the diagnosis,treatment and prognosis was analyzed.Results The intracranial pressure monitoring group showed a significantly shorter period concerning osmotic dehydration [(14.24 ± 7.93) days vs (21.61 ± 11.97)days,P＜0.01],ICU stay [(14.38 ±7.56)days vs (24.71-± 17.94)days,P＜0.01] and total hospital stay [(17.20 ±8.09)days vs (33.92 ± 21.70)days,P＜0.01] as well as a better GOS [(4.15 ± 1.22) points vs (3.69 ± 1.56) points,P ＜ 0.05],as compared with group without intracranial pressure monitoring.Conclusions Craniotomy,especially decompressive craniectomy,is one of the most important treatment means to control cranial pressure and ensure cerebral perfusion pressure in patients with bifrontal contusions (in particular the moderate and severe ones).Besides,intracranial pressure monitoring is conducive to selection of surgery timing and is instructive to combined treatment,such as osmotherapy,intracranial pressure controlling and assurance of cerebral perfusion pressure.

Four kinds of colophony were used to isolate the herbicidal substance from the toxin produced by isolate PAM1 of Pythium aphanidermatum, which had herbicidal activity to Digtaria sanguinealis L and the results showed X-5 was the best one for absorbing herbicidal substance among the four kinds of colophony used. Four solvents including 50% ethanol, 95% ethanol, ethyl acetate and acetone were used as eluting solvent and 2 procedures were tested, and the results of growth inhibition and seed germination indicated that the best eluting procedure was procedure 1.