Objective:To construct the expression vector pSuper.Hsp90?/pSuper for further research of the Hsp90?.Methods: In accordance with the Hsp90? sequence in gene bank,three sequences respectively targeting Hsp90? were cloned into the RNA expression vector pSuper.Results:It was confirmed by enzyme digestion,and electrophoresis that the construction of the recombinant expression vector pSuper-Hsp90?1,pSuper-Hsp90?2 and pSuper-Hsp90?3 was successfu1.pSuper-Hsp90?1 and pSuper-Hsp90?2 were successfu1 by sequencing.The expression of Hsp90? mRNA in HEK293 cells with pSuper-Hsp90?1,pSuper-Hsp90?2 and pSuper-Hsp90?3 decreased,and pSuper-Hsp90?1 notably decreased.Conclusion:The recombinant plasmid against Hsp90? RNAi can be successfully constructed in vitro,and pSuper-Hsp90?1 is the possible optimal target.

Objective: To observe the influence of electroacupuncture on depression and defects of neurological functions in the patients with post-stroke depression. Methods: 90 cases of the apoplectic patients confirmed clinically by CT or MRI were selected and divided randomly into three groups by digital table, 30 cases in each group. All three groups of the patients were treated for wind stroke, with electric stimulation added for the acupuncture group,Fluoxetine added for medication group, and placebo added for control group, continuously for 60 d. Results: Compared to the control group, the scores in Hamilton Depression Rating Scale (HAMD) and Modified Edinburgh-Scandinavia Stroke Scale (MESSS) decreased in the electroacupuncture group and medication group, and the scores in Barthel index increased (P＜0.05). The general therapeutic effects were better in the electroacupuncture group and medication group than in the control group, and the general therapeutic effects were similar in the electroacupuncture group and medication group. Conclusion: Eiectroacupuncture can improve depressive behavior and promote rehabilitation of the nervous functions.

Objective To explore ffie effects of magnesium isoglycyrrhizinate injection,polyene phosphatidylcholine(PPC)and reduced glutathione(GSH)in patients with liver cirrhosis accompanied by benign obstructive jaundice after surSery.Methods Fifty-nine patients with liver cirrhosis and benign obstructive jaundice caused by gallstones were selected,and all of them were treated with magnesium isoglycyrrhizinate injection and GSH preoperatively.Thirty-one odd number patients received magnesium isoglycyrrhizinate injection,PPC and GSH treatment(group A)after surgery immediately,28 even number patients still continued to be treated with magnesium isoglycyrrhizinate injection and GSH(group B).The plasma alanine aminotransferase(ALT),aspartate aminotramsferase(AST),total cholesterol(TBIL)and cholinesterase(ChE),and prealbumin at the time of 1,3 and 7 day after surgery were compared between the two groups.Results Seven patients were excluded from this research including 3 cases in group A,4 cases in group B.The plasma ALT,AST,TBIL of the group A at postoperative 3,7 day[(193 ± 48)and(63 ± 17)U/L,(194 ± 57)and(54 ± 19)U/L,(99 ± 20)and(28 ± 9)μmol/L]were significantly lower than those of the group B[(235 ± 50)and(103 ± 33)U/L,(235 ± 64)and(82 ± 22)U/L,(127 ± 45)and(43 ± 18)μ mol/L,respectively](P＜ 0.05),whereas the plasma ChE and prealbumin of the group A[(3781 ± 294)and(4405 ± 469)U/L,(0.22 ± 0.07)and(0.30 ± 0.04)g/L]were significantly higher than those of the there were obvious differences of the plasma ChE and prealbumin at postoperative 3 or 7 day compared with those at postoperative 1 or 3 day in the group A(P ＜ 0.05).But this were improved at postoperative 7 day compared with those at postoperative 3 day in the group B(P＜ 0.05).Conclusion Combined treatment of magnesium isoglycyrrhizinate injection,PPC and GSH can be more effective to reduce the injury of the liver cells and promote the recovery of liver function for patients with liver cirrhosis and obstructive jaundice.

Objective:Combining the reality of school, to explore the method of training on the implementation of the outstanding doctor education.Methods: Deepening the reform of teaching, curriculum integration, constructing reasonable curriculum system, strengthen the innovation ability and practice ability, build a outstanding doctor training system, revision of the talent training scheme, the implementation of outstanding physician education program.Results:in our school outstanding doctor the actual situation of the education training, and further analysis and research, explore the method of training on the implementation of the outstanding doctor education.Conclusion: by deepening the reform of teaching, constructing curriculum system, strengthen the innovation ability and practice ability, build a outstanding doctor training system, revision of the talent training scheme, the implementation of outstanding physician training plan, so the method of training on the implementation of the outstanding doctor education is feasible.

Objective To evaluate the impact of comorbidity and age on the tolerance of firstline single-agent chemotherapy in elderly patients with advanced non-small cell lung cancer(NSCLC).Methods Clinical data of 61 elderly patients with advanced NSCLC(aged over 70 years,median age 72 years) receiving first-line single agent chemotherapy were retrospectively analyzed in this study.Performance status(PS) between 0-1 score was in 52 patients,PS 2 score in the other 9 patents.Patients were treated with gemcitabine or docitaxel as the first line chemotherapy,and the median number of chemotherapy cycles was 3.4.Comorbidity was assessed by Charlson comorbidity index (CC1).Patients with CCI equal to 0 were classified as non comorbidity group(n=26),and patients with CCI≥1 were classified as comorbidity group(n=35).Adverse reactions were graded by using the criteria of NCI-CTC v3.0.Results Age and PS could not predict adverse effects of grade 3 or 4.The incidence of hematologic toxicity of grade 3 or 4 was higher in comorbidity group than in noncomorbidity group(40.0％ vs.15.4％,x2 =4.36,P=0.037).The incidences of febrile neutropenia,non hematologic toxicity of grade 3 or 4 and treatment suspension were higher in comorbidity group than in non-comorbidity group.The most common types of comorbidity were diabetes and chronic pulmonary disease.The incidence of non-hematologic toxicity of grade 3 or 4 was increased in patients with chronic pulmonary disease as compared with patients without chronic pulmonary disease(41.4 ％vs.11.5％,x2=6.061,P=0.032).Conclusions The incidences of adverse reactions,especially hematologic toxicity of grade 3 or 4 are significantly increased in patients with comorbidity after singleagent chemotherapy.Evaluation of comorbidity before treatment is helpful to predict the tolerance of single-agent chemotherapy in elderly NSCLC patients.

BACKGROUND:Transient receptor potential protein channel is distributed in many kinds of tissue and cells in human body, and takes part in many physiologic functions and pathological changes of human body. However, up to now, transient receptor potential melastatin 8 (TRPM8) has not been reported in human cartilage.
OBJECTIVE:To study the expression of TRPM8, a subfamily member of transient receptor potential gene, in human knee joint cartilage.
METHODS:Fifteen samples were selected from patients after amputatioin due to severe trauma. Total protein was extracted from 300 mg cartilage tissue of every patient. To detect the distribution of TRPM8 in human
cartilage, cartilage sections were col ected in the same region. The expression and distribution of TRPM8 in human cartilage was detected by western blot analysis and immunofluorescence respectively.
RESULTS AND CONCLUSION:Western blot analysis showed that specific TRPM8 band was visible at molecular weight of 130 000, and as the age increased, TRPM8 expression in human cartilage was gradual y decreased. Immunofluorescence staining results showed that TRPM8 was evenly located in the cellmembrane of chondrocytes. TRPM8 is expressed in the human knee cartilage, specifical y in member of chondrocytes.

Objective To investigate the characteristic changes in the mRNA and protein of complement 3 in spinal dorsal horn of rats with neuropathic pain,and the role of abnormal activation of complement protein in the mechanism of pain production.Methods Eighty-four healthy male SD rats were divided randomly into seven groups(n=12 for each group):normal control group,sham-operation 1d,3d and 7d groups,and chronic constriction injury of the sciatic nerve(CCI)1d,3d and 7d groups.The left sciatic nerve was ligated loosely in CCI groups,while it was only exposed but not ligated in rats in sham-operation groups.The mechanical and thermal pain thresholds were measured on 1,3 and 7 days after operation,and the mRNA and protein of complement 3 in the dosal horn of the spinal cord were determined respectively by RT-PCR,immunoturbidimetry and immunohistochemistry.Results The mechanical and thermal pain thresholds were observed to be lowered in rats one day after the sciatic nerve ligation,and a state of hyperalgesia was found to be persistent up to 7 days after CCI.This symptom was not observed in sham operation group.The expression of mRNA and protein of complement 3 in spinal dorsal horn were increased on 1,3 and 7 days after CCI.Interestingly,a high expression of mRNA of complement 3 was also observed in rats one day after sham-operation.Both mRNA and protein of complement 3 were not obviously elevated in rats of sham operation 3d,7d groups and normal control group.Conclusion The mRNA and protein of complement 3 in spinal dorsal horn are highly up-regulated in rats with neuropathic pain,suggesting that the characteristic dynamic changes in complement may contribute to the establishment and maintenance of hyperalgesia.

Objective To compare the in-hospital and follow-up clinical results of percutaneous coronary intervention(PCI)and coronary artery bypass grafting(CABG)in patients with coronary artery disease and diabetes mellitus.Methods The DESIRE(drug-eluting stent impact on revascularization)is a single-center registry comparing coronary revascularization in our institution before and after the introduction of drug-eluting stents.Between July 2001 and June 2002,July 2003 and June 2004,1040 patients were referred for revascularization(PCI or CABG)during hospitalization.Clinical,angiographic,revascularization data clinical condition in hospital and the follow-up results were collected for retrospective analysis.In the present study,patients with diabetes were selected and divided into PCI and CABG groups.The primary endpoint is in-hospital and follow-up major adverse cardiovascular and cerebral events(MACCE).Results Compared with the CABG group,the PCI group had lower in-hospital MACCE rates(P0.05),but the PCI group had higher rates of repeat revascularization than that of the CABG group(P0.05).Conclusion Although PCI has lower in-hospital MACCE rates in diabetic patients,the follow-up MACCE rates of PCI and CABG are comparable,and PCI has even higher rates of repeat revascularization than CABG.With the increasing use of drug eluting stents,PCI might have better performance.

ObjectiveTo observe the effect of family rehabilitation plan (FRP) on motor function of stroke patients with hemiplegia.Methods64 patients were randomly divided into the FRP group (32 cases) and control group (32 cases). The patients in the FRP group critically followed the schedule and activities scheduled on FRP and were assessed and guided in out-patient department every two weeks. The patients in the control group made schedule and carried on activities by themselves. Motor function was assessed with Fugl-Meyer Assessment (FMA) and modified Barthel index (MBI) respectively.ResultsMotor scores of the patients in two groups significantly increased after treatment ( P<0.001), and that of the FRP group was significantly better than that of the control group ( P<0.001).ConclusionThe FRP can significantly improve motor function of stroke patients with hemiplegia.

Objective To study the change and significance of osteoprotegerin(OPG)/receptor activator of NF κB ligand(RANKL)during the process of rat arterial smooth muscle cells (SMC) to differentiate subgroups:7 d (early) subgroup and 14 d (late) subgroup.into osteoblast-like cells.Methods The rat arterial SMCs were divided randomly into the SMC group,atorvastatin group,and osteoblast-like group.Each group was also divided into 2.The osteoblast-like group was given β-glycerophosphate and vitamin C in culture medium,the atorvastatin group was given both β-glycerophosphate,vitamin C and atorvastatin.Von Kossa staining,Ca2+ content assay,ALP activity assay and osteocalcin assayed with Western blot were used to check the level of calcification.Real-time PCR was used to check the mRNA expressions of OPG and RANKL.Results There was high expression of OPG (early 2.71 ±0.08,late 2.69 ±0.02) but no RANKL in the SMC group all the time.The expressions of OPG were increased in the early subgroup and decreased in late subgroup in the atorvastatin group (early 3.52±0.05,late 2.50±0.03) and osteoblast-like group (early 4.18±0.10,late 2.30 ± 0.11).And the expressions of RANKL were both increased in the atorvastatin group and osteoblast-like group dnring the calcification process (from 1.01 ± 0.19 to 2.40 ± 0.10,and from 1.70±0.07 to 3.22±0.11,respectively).Among the three groups,the ratio of OPG/RANKL was decreased with the increasing calcification level (F =52.93,2.33,both P＜0.05).And compared with the early subgroups,the ratio of OPG/RANKL in the late subgroups was also decreased along with the increase of calcification level in each group(F=38.71,1.74,both P＜0.05).Conclusions The ratio of OPG/RANKL has a negative correlation with the level of the osteoblast-like cells differentiation,and atorvastatin could inhibit the calcification.