OBJECTIVE:To provide reference for standardized management of national drug standard in China.METHODS:Special inspections about the implementation of national drug standard were performed in pharmaceutical manufacturers of Deyang area.Based on the results of daily inspections,national drug standard was analyzed and discussed comprehensively.RESULTS:National drug standards were too many and diverse so that they influenced standardized management and implementation,which involved several aspects such as legality and seriousness of the management of drug standard,the conformity of relevant policy with drug standard,the standard of raw material and excipients,standardization of management approach for reference sample.CONCLUSION:Rigorous,complete and accurate national drug standards should be formulated,and relevant laws and regulations should be improved.Pharmaceutical enterprises can be urged to establish standardized drug standard file and to organize production and testing in accordance with national drug standard.

39 cases of fatally abused children were collected during April,1984 to febraury,1987 in Los Angeless,U.S.A.73 cases of non-abused children were obtained as control.Both eyes of each case were taken and examined histo- pathologically.The intraocular changes occured in 64% of the fatally abused children.They were:Retinal hemorrhage,vitrous hemorrhage,retinal de- tachmedt,papilledema,subdural hemorrhage of the optic nerve and traum- atic cataract.On the other hand,the intraocular changes occured in 28% of control cass,especially in those of traffic and other accidents which usually caused the injuries of head and neck.Considering the intraocular changes and the general injuries together,we suggest that the most important mec- hanism of the intraocular changes be head injuries and whiplash syndrome.

To evaluate the effectiveness of comprehensive psychological intervention for reducing the preoperative stress response in sterility women. Forty sterility women were equally divided into intervention group and control group. Heart rate(HR), blood pressure(BP), Zung anxiety grade and depression grade scores(SAS, SDS) were recorded; serum concentrations of glucose (GLU), cortieosteroid (Cor), adrenocorticotropic hormone(ACTH) and angiotensin (A Ⅱ) were measure in both groups at admission and before operation. These parameters before surgery were significant higher than those at admission for control group(P<0.05); but there were no significant differences for intervention group (P>0.05). The results indicate that comprehensive psychological intervention can reduce the preoperative stress response of sterility patients.

In order to explore the significance of cardiomyocyte apoptosis in the early myocardial ischemia,the rat model of acute myocardial ischemia was established,and the apoptotic cells were detected in the early phase of ischemia(within 6h)with TUNEL method.The results showed that scanty apoptotic cells could be observed in the ischemic region 1h after ischemia,and reached the peak 3h after ischemia,then decreased.No apoptotic cells were found in the normal region.In the peri ischemic region,apoptotic cells could also be observed 1h after ischemia,and reached the peak 5h after ischemia.It is indicated that apoptosis is the major form of early ischemic myocardial damage,and the detection of apoptotic cardiomyocyte may provide a new sensitive and objective method for the postmortem diagnosis of early myocardial infarction.

To study the expression of both Fos and HSP70 proteins in the early stage of acute myocardial ischemia/reperfusion(I/R) in rats with immunohistochemistry in order to search the objective morphologic evidence for the forensic pathological diagnosis.I/R rat model was established by ligation of the anterior branch of the left coronary artery.All animals were divided into 7 groups.Tissues were cut and stained immunohistochemistry with the corresponding anti Fos and anti HSP70 sera as the first antibodies.There was no expression of both Fos and HSP70 proteins in control group.The expression of HSP70 protein began to increase in myocytes 1h after I/R in the ischemic areas and increased gradually wtih the prolongation of ischemia,while it began to express 2h after I/R in the non ischemic areas.Fos protein began to express 0 5h after I/R in the ischemic areas,1h in the non ischemic areas.The expressions of both HSP70 and Fos proteins in the non ischemic areas were weaker than those in the ischemic areas.Meanwhile,the expression of HSP70 and Fos proteins appears firstly in the inner layer of myocardium and extended gradually toward the outer layer.The expression of Fos protein mainly located in the inner layer of myocardium 0 5h after I/R and then extened to the whole layer of myocardium 1h after I/R.The expression of Fos protein in outer layer was stronger than that in inner layer at 4h after I/R.The expression of HSP70 protein located mainly in the inner layer of the myocardium at 1h and 2h after I/R,and then extended to the whole layer of the myocardium at 4h and 6h after I/R.The results indicated that the I/R induced the expression of both HSP70 and Fos proteins in the early stages of myocardial ischemia.The location and the intersity of the immunoreactivity of these two proteins changed at the different stages after I/R.These changes observed in the present study might be useful for the forensic pathological diagnosis of the early myocardial ischemia.

BACKGROUND:There is lack of an objective and standardized animal model for assessing the therapeutic effect of physical and medication treatment on bone defoct, the effectiveness of operation, as well as the role of bone substitute in the repairing of bone defects.DESIGN:Verified study on the experimental model of bone nonunion in rabbitsSETTING: Department of Orthopaedics in Shenzhen people' s Hospital MATERIALS:Twenty common grade pure New Zealand rabbits of either gender were selected with body mass of (2.5±0.5)kg,aged 6 to 8 months.METHODS :This experiment was carried out at the experimental animal center of Shenzhen people's Hospital between May and August 1999. 1.5cm bone segment (including periosteum)was cut off in the middle of forearm radius in 20 common grade pure NewZealand rabbits,the broken ends were covered with bone wax, 10 weeks later, the bone nonunion status was assessed by macropathological observation, pathohistological and X-ray examination.MAIN OUTCOME MEASURES:Observations on rabbit forearm radius defects by macropathological observation, pathohistological and X-ray examination.RESULTS :Twenty rabbits(40 side radius)were enrolled in this study and weeks later, bone defect region was found filled with fibrous cicatricial tissue without osseous connection ,bone wax was not absorbed, capitulum was ossified with medullary cavity blocked,a small amount of callus formed at both broken ends of fractural bone ,length of bone defect ranged from 0.8 to blocked under optical microscope,chondrocyte and osteocyte could be observed arranging disorderly and covered with fibrous membrane,defect reosseous connection could be detected at defect region at week 10,broken end was ossified and medullary cavity was blocked ,there was small amount of callus appeared at both broken ends displaying irregular shape.CONCLUSION:Bone nonunion experimental animal was successfully established on rabbits in this study, with pathological changes meeting the need of bone nonunion and displaying typical properties,which can be used as reliable and feasible experimental animal model.

BACKGROUND: The study of isolation, purification, culture, cell labeling, inducing factors, effects of gene transfection on cytobiology, cell carrier construction, and time window for back transplantation of cell compound pertaining to marrow stromal cells (MSCs) is still in its infancy. OBJECTIVE: To search for an in vitro culture method that can be simply and effectively obtained. DESIGN, TIME AND SETTING: The present cytological in vitro experiment was performed at the Beijing Institute of Genome, Chinese Academy of Sciences between June 2006 and July 2007. MATERIALS: Eight specific pathogen-free New Zealand rabbits, aged 6 weeks, were provided by the Laboratory Animal Center, Institute of Genetics and Development, Chinese Academy of Sciences. METHODS: Under sterile condition, 1 mL rabbit bone marrow was taken and diluted with D-Hanks solution. Following centrifugation and subsequent supernatant removal, bone marrow was re-suspended using dulbecco's modified eagle's medium (DMEM) for single cell suspension. Next, single cell suspension was dropped onto the liquid surface of equal-volume lymphocyte separation medium (density: 1.077). Subsequent to centrifugation, cloudlike mononuclear cell layer was collected and re-suspended with DMEM containing 20% fetal bovine serum. The cells were inoculated at lxl0/cm2 and purified by adherent method. When 70%-80% of flask bottom was covered, cell digestion and passage was performed.MAIN OUTCOME MEASURES: Cell growth was observed with an operating microscope. Surviving cells were counted by Trypan blue viability test. Cell identification was performed by hematoxylin-eosin staining. Through the use of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, cell viability was detected to observe the cellular resuscitation of the cultured cells following cryopreservation. RESULTS: Twenty-four hours after inoculation, cells began to adhere to the wall, exhibiting short shuttle- or triangle-shaped appearance with different sizes of cellular processes. Three days later, adherent cells began to divide, and cell clusters could be found in some areas; One week later, most of cells exhibited scattered fibroblast-like growth; After passage, cells were evenly distributed with long shuttle-shaped appearance and arranged orderly. Following 3 passages, there wound be 5×106-1×107 adherent cells in 1 mL MSC suspension. Approximately 90% of MSCs survived and identified as mononuclear cells. Cells vigorously grew at days 1-6 after inoculation and reached a peak level at day 8, followed by a viability decline. After 56 days of resuscitation, frozen cells displayed a good and rapid growth. CONCLUSION: Highly purified MSCs can be acquired by gradient centrifugation of lymphocyte separation medium. Enough seeded cells can be obtained by in vitro culture and the cellular viability does not change after frozen preservation and resuscitation.

BACKGROUND: SOX-9 plays an important role in occurrence and development of cartilaginous tissues,enhances agglutination of mesenchymal cells,has structural domains of transcriptional activation,and can directly regulate the transcription,OBJECTIVE: To construct pDC316-SOX-9 plasmid for transfection of rabbit bone marrow stromal cells (BMSCs) using SOX-9 gene,and to investigate the effects of SOX-9 gene on growth characteristics of BMSCs and product expression. DESIGN,TIME AND SETTING: The cell gene engineering in vitro experiment was performe,d at the Beijing Institute of Genome, Chinese Academy of Scienees from June 2006 to January 2007.MATERIALS: Eight SPF New Zealand rabbits aged 6 weeks were used for culture of BMSCs.METHODS: pDC316-SOX-9 plasmid was used for transfection of rabbit BMSCs by liposome mediated method,Cell transfection included a SOX-9 plasmid transfection group,a blank plasmid group and a blank control group (only treatment of liposome). MAIN OUTCOME MEASURES: Cell morphology; growth activity; The SOX-9 protein expression in rabbit BMSCs were detected by immunohistochemistry,hematoxylin-eosin staining,reserve transcriptase-polymcrase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). RESULTS: Some cell colonies were detected at 4 days after pDC316-SOX-9 plasmid transfection.Spindle-shaped cells were collected after clone amplification.Cells in the blank control group gradually died over time.Cell activities in the SOX-9 plasmid transfeetion group and the blank plasmid group significantly prolonged,reached a peak at 2 weeks of transfection,and then gradually decreased.At 6 days,BMSCs were yellow in the SOX-9 plasmid transfection group following immunohistochemistry,expressing SOX-9 protein.Hematoxyliu-eosin staining showed many dikaryocytes,rich cell proliferation,similar to chondroblasts.No SOX-9 protein expression and unproductive cell proliferation in the blank plasmid group.SOX-9 mRNA was detected in the SOX-9 plasmid transfection group,but SOX-9 mRNA was not detected in the blank plasmid group and blank control group.SOX-9 levels were significantly higher in the SOX-9 plasmid transfection group than in the blank plasmid group and blank control group at 24,48 and 72 hours,1,2 weeks (P＜ 0.01).CONCLUSION: Rabbit BMSCs were successfully transfected with pDC316-SOX-9 plasmid to enhance cell growth activity and to persistently stably secrete SOX-9 protein,resulting in the differentiation of BMSCs into cartilages.

Objective To observe the morphologic changes of of vascular buds in vertebral cartilage endplate in age-specific rabbits and also to investigate the correlation between the changes of vascular buds and interverbral disc degeneration. Methods There were 15 New Zealand white rabbits in our study,which include three groups, 2-week-old rabbits, 1-year-old rabbits and 3-year-old rabbits, and each groups had five rabbits. The X-ray radiograph, histology and scanning electron microscope were used to observe the changes of vertebral cartilage endplate. According to Miyamoto standard, the interverbral disc was graded 1-5, and scored 1-5 respectively. Results The changes of micro-vascular structure of vertebral cartilage endplate were observed during aging. Under the scanning electron microscope, the vascular structure degenerated gradually, and disappeared in the end. The blood vessels in the central region of the vertebral cartilage endplate reduced more obviously than those in periphery region. The severe degeneration was found in vertebral endplate, compared with intervertebral disc. The changes of vascular buds in rabbits vertebral cartilage endplate had positive correlation with the vertebral endplate calcification and the interbertebral disc degeneration. Conclusion Changes of vascular buds in vertebral endplate may accelerate intervertebral disc degeneration.

Background: Chronic obstructive pulmonary disease (COPD) is an important clinical disease, and its global prevalence and mortality rates are high. It is meaningful to investigate the efficacy of integrative respiratory rehabilitation training in quality of life and respiratory physiology of COPD patients in stable phase. Objective: To observe the efficacy of integrative respiratory rehabilitation training in exercise ability and quality of life of COPD patients in stable phase. Design, setting, participants and interventions: Eighty outpatients and inpatients with COPD from Department of Respiratory Medicine, Taihe Hospital, Yunyang Medical College were randomly divided into 4 groups, with 20 patients in each group. The patients in group A only received drug therapy, the patients in group B received traditional qigong training, the patients in group C received modern rehabilitation training, and the patients in group D received integrative respiratory rehabilitation training. Main outcome measures: Chronic respiratory questionnaire (CRQ), 6-minute walking distance and Borg score in each group were examined before and after one-, three-, and six-month and one-year treatment. Results: The 6-minute walking distance, Borg score and CRQ score in group A had no significant changes after treatment (P>0.05). After one-month treatment, there were no significant differences in 6-minute walking distance and Borg score in groups B, C and D as compared with those before treatment (P<0.05). After three-month treatment, 6-minute walking distance and Borg score were improved in groups B, C and D (P<0.05). After six-month and one-year treatment, 6-minute walking distance, Borg score and CRQ score in groups B, C and D were improved as compared with those before treatment (P<0.05), and there were significant differences between group D and any of groups A, B and C (P<0.05). Conclusion: Modern rehabilitation training, traditional qigong training and integrative respiratory rehabilitation training programs all can improve the quality of life and exercise ability of COPD patients, and integrative respiratory rehabilitation training program is better than modern rehabilitation training and traditional qigong training programs. The efficacy of respiratory rehabilitation training is time-dependent, and need long-time adherence to the therapy.