Aplastic anemia(AA)is a rare disease oringed from hematopoietic cell,characterized by failure of the bone marrow and pancytopenia in peripheral blood.The mechanism of AA is indistinct,it can be congenital,or acquired disposition.Acquired AA may be secondary to poisonous,radiation,virus infection and some medicine.All these factors can injure bone marrow directly,suppressing the proliferation and differentiation of bone marrow,on the other hand,the immune system play a pivotal role indirectly.This review focused on the pathologic classification,mechanism of drug-induced aplastic anemia.

BACKGROUND: The glial scar formation after spinal cord injury in mammals is the physical and chemical barriers for neural regeneration, and relieving or delaying glial scar formation can provide benefit conditions for the regeneration of injured spinal cord.OBJECTIVE: To design and screen short hairpin RNA (shRNA) interfere molecular targeting the gene coding region of glial fibrillary acidic protein (GFAP) in rat, and reconstruct the eukaryotic vector of shRNA.DESIGN: An observational animal experiment.SETTING: Department of Spine Surgery, Shenzhen Hospital affiliated to Southern Medical University.MATERIALS: Twenty-five Wistar rats of clean degree, either male or female, weighing 20-25 g, were used. DMEM/F12,lipofectamine2000, Trizol RNA separating kits); fetal bovine serum (Hyclone); BamH Ⅰ, HindⅢ, Pstl, Sail and T4 ligases;Plasmid mini preparation kit and DNA gel extraction kit.METHODS: The experiments were carried out in the Shenzhen Hospital affiliated to Southern Medical University from October 2005 to June 2006. Three pairs of shRNA template which composed of 19 bp reverse repeated motif of GFAP target sequence with 9 bp spacer were designed and synthesized, then they were inserted directionally into plasmid Psilencer 2.1 respectively to generate small interfering RNA (siRNA) eukaryotic expression vector. ShRNA molecules were transfected by liposome via ex vivo expression repressive model of GFAP of rat spinal astrocytes. The effects of expressive suppression were detected with real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, and then the optimal shRNA eukaryotic vector of repressive expression of GFAP was screened.MAIN OUTCOME MEASURES: ① Interfering sequence specific shRNA template synthesis; ② Constructing specific recombinant plasmid eukaryotic expression vector. ③ Culturing rat spinal astrocytes in vitro; ④ Effects of expressive suppression on GFAP in primary astrocytes after RNA interference detected with real-time fluorescence quantitative RT-PCR and Western blot.RESULTS: Sequence analysis showed that GFAP-shRNA recombinant plasmid eukaryotic expression vector was successfully constructed, and optimal GFAP-shRNA eukaryotic vector was screened using real-time fluorescence quantitative RT-PCR and Western blot. The GFAP expressions were reduced by 81%, 63% and 56% at the levels of mRNA and protein respectively.CONCLUSION: GFAP-shRNA eukaryotic expression vectors were successfully constructed and screened. The gene expression GFAP of primitive rat astrocyte can be suppressed significantly by the GFAP-shRNA eukaryotic expression recombinant optimized via ex vivo cellular expression suppression model, which should pave the way for the following multiple targets of RNAi genetic manipulation in the treatment of suppression of glial scar formation after spinal cord injury.

Now the in-service staff have been permitted to apply for master's degree with the same educational level aspostgraduates. This is a signal reforming policy in postgraduate education and degrees management area in China. Manyhigher professionals are trained for our socialist modernization construction by this education technology. Although therelatively perfect system to manage this project was established in China, there are still some problems demandingprompt solution on the management thoughts, management process as well as management methods in some authorizedunits.

OBJECTIVE:To introduce the Fair PharmaCare program of BC Canada for the reference of the reform of China's medical insurance system.METHODS:The running procedure of Fair PharmaCare program of BC Canada was introduced in detail and its characteristics were analyzed.RESULTS & CONCLUSIONS:Fair PharmaCare program covers all BC residents and protects them against economic difficulties in pharmacare.The program serves as a reference for the reform of China's medical insurance system.

Objective To investigate the distribution of complications and the relationship with neurologic subtype and gross motor function in preterm infants with cerebral palsy (CP). Methods The type, grade of Gross Motor Function Classification System (GMFCS), intelligence,speech, ophthalmologic consultation, brainstem auditory evoked potential and electroencephalogram of 135 preterm infants with CP were reviewed. Results There were 284 complications in total, (2.10±1.33) per child, and was significantly different among various types of CP (F=5.50, P<0.001). The incidence of mental retardation and speech disorder was significant different among various types (P<0.05). The incidence of mental retardation, speech disorder, visual impairment and epilepsy increased significantly (P<0.05) in spastic quadriplegia infants,compared with those with diplegia and hemiplegia. The incidence of mental retardation, speech disorder, visual impairment and musculoskeletal disorder was significantly different (P<0.05) among various grades of GMFCS. The frequency of complications was more in children unable to walk (GMFCS Ⅳ~Ⅴ) than able to walk (GMFCS Ⅰ~Ⅲ) for children over 2 years old (t=70.05, P<0.001). Conclusion The incidence of mental retardation, speech disorder, visual and hearing impairment, secondary musculoskeletal disorder and the multiple disorders are related with neurologic subtype and/or the grade of GMFCS.

BACKGROUND: It is demonstrated that bone marrow stem cells (BMSCs) can generate neurosphere structures, which is similar to cloning sphere of neuron-specific enolase (NSE), in a specially induced system in vitro; therefore, BMSCs draw more and more attention as seed cells to repair central nerve injury.OBJECTIVE: To investigate the differentiation from BMSCs into neuron-like calls induced by fetal spinal cord tissue.DESIGN: Observational study.SETTING: Department of Spine Surgery, Second People's Hospital of Shenzhen.MATERIALS: SD rats (16 pregnant days old and 2 months old) were provided by Animal Center, Tongji Medical College,Huazhong University of Science and Technology. Single antibody of NSE, multi-antibody of glial fibrillary acidic protein (GFAP) and single antibody of neurofilament (NF200) were provided by Wuhan Boster Company.METHODS: The experiment was carried out in Immune Opening Laboratory and Central Laboratory, Basic Medical Department, Tongji Medical College, Huazhong University of Science and Technology in September 2006. Bones of lower extremities of rats were collected to centrifuge BMSCs. Fetal spinal cord tissue homogenate was extracted from 16 pregnant days old rats to make inducing solution and induce differentiation of BMSCs. Otherwise, embryo muscle tissue was used to make muscle tissue homogenate as the same way so as to regard as the controls.MAIN OUTCOME MEASURES: BMSCs underwent morphological observation after induction; in addition, anti-NSE, NF200 and anti-GFAP were used to label neurons and astrocytes, respectively. Ten non-overlapping sights were randomly selected from positive reactive-induced cells after immunohistochemical staining under optic microscope to calculate ratio of positive cells of NSE and NF200 counting for total numbers of cells.RESULTS: ① Morphological changes of BMSCs after induction: During early induction, optic microscope indicated that soma of partial cells rebounded; whose apophysis was long and thin; apophysis of differentiated cells grew gradually and intercrossed each other. It was similar to nerve cells and some branches were similar to dendrite branches. However,morphological changes of cells in the control group were not obvious. ② Expression of relevant antigens differentiated from BMSCs into neuron-like cells at one week after induction: Most cells in spinal cord homogenate group expressed as positive NSE and NF200; a few of cells expressed as GFAP. While, positive staining of nerve cell antibody was not observed in the control group; meanwhile, positive reaction of nerve cell antigen was not observed in the control group,too. Immunohistochemistry examination demonstrated that positive rates of NSE and NF200 expressions were (68±1.7)%and (76.2±2.9)%, respectively.CONCLUSION: Fetal spinal cord tissue homogenate can induce differentiation from BMSCs into neuron-like cells.

Objective To observe the concentration effects of alcohol in perineurial block on the structure and function of sciatic nerve and it's innervated muscles in rats so as to provide the basis for clinical application of chemical neurolysis.Methods One hundred and fifty female Sprague-Dawley rats were used and randomly assigned into a blank group(BG,n =6),a control group(CG,n =36),a 50％ alcohol group(50G,n =36),a 75％ alcohol group(75G,n =36)and a 99.9％ alcohol group(99.9G,n =36).The CG received physiological saline injection,the 50G,75G,99.9G received corresponding concentrations alcohol perineurial block,respectively.Changes of motor function was assessed,electrophysiological and histomorphological observations of sciatic nerve and its innervated muscles were conducted before and at 24 h,72 h,1 week,4 week,12 week after block.Results(① Hypokinesia and decrease of motor conduction velocity(MCV)were observed at 24 h after block,and peaked at 72 h after block; at 1 week after block,the motor function and MCV improved,the improvement persisted to the 12th week (P ＜ 0.05); but at the 12th week,MCV was still slower than that before block(P ＜ 0.01).②There were significant differences with regard to motot function and MCV of sciatic nerve among 99.9G and the other groups at e ery time point after block(P ＜0.05); ③ Reversible dengeneration of sciatic nerve and hind limb muscle,nonreversible necrosis of muscle occurred more seriously with the increase of alcohol concentration,and the cicatrization in 99.9G was more obvious than that in 50G and 75G at the 12th week; ④Structural lesion of sciatic nerve occurred at 24 h after block and peaked at the 72th h,myelinated axonal sprouts appeared at 1 week after block and persisted to the 12th weck.Conclusions ① The effects of 99.9 ％ alcohol perineural block on the structural lesion,motor function and MCV of sciatic nerve and its related muscle were more obvious than those of 50％ and 75％ alcohol; ② The destructive effect of alcohol block would maintain more than 12 week and neural restoration would maintain more than 12 weeks too.

AIM To establish an HPLC-DAD method for the simultaneous content determination of four constituents in Compound Kendir Leaves Tablets Ⅰ (Apocyni veneti Folium,Chrysanthemi indici Flos,Stephaniae tetrandrae Radix,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Shimadzu VP-ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of methanol-0.5％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 260 nm and 325 nm.RESULTS Chlorogenic acid,hydrochlorothiazide,buddleodide and promethazine hydrochloride showed good linear relationships within the ranges of 24.91-498.2 ng (r =0.999 9),286.33-5 726.7 ng (r =0.999 9),10.04-200.9 ng (r =0.999 9) and 154.80-3 096.1 ng (r =0.999 9),whose average recoveries were 98.3％ (RSD =1.3％),99.1％ (RSD =0.6％),98.5％ (RSD =1.0％) and 99.3％ (RSD =1.2％),respectively.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Compound Kendir Leaves Tablets Ⅰ.

Objective To compare the clinical efficacy of single and double locking compressing plates(LCP)treatment for Schatzker type Ⅵ tibial plateau fractures.Methods Patients(n=90)with Schatzker type Ⅵ tibial plateau fractures were randomly divided into two groups according to the treatment methods (single LCP group and double LCP group;45 patients in each group) and analyzed retrospectively.Operative duration,intraoperative bleeding volume,postoperative wound infection,Rasmussen anatomical score and functional score were measured and compared between the two groups.Results More favorable results were achieved in the single LCP group than in the double LCP group in operative duration [(51.4 ± 4.03)h vs.(76.2±5.5)h(t=24.514,P＜0.001)]and intraoperative bleeding volume[(173.7±8.8) ml vs.(212.4 ± 4.9) ml (t =25.911,P ＜ 0.001,respectively)].Further more,three cases of postoperative woundin fection were seen in the double LCP group,while none was observed in the single LCP group.There was no significant difference in the percentage of satisfactory Rasmussenanatomical scores and functional scores between the two groups one year afteroperation(~ =-0.442,P=0.659).Conclusions Compared with double LCP fixation,single LCP fixation is more advantageous for the treatment of Schatzker type Ⅵ tibial plateau fractures.