Objective To explore whether the polymorphisms within the tumor necrosis factor alpha (TNF-?) and TNF-? gene are associated with the clinical types of patients with chronic hepatitis B virus (HBV) infection. Methods By using a restriction fragment length polymorphism (RFLP) assay of polymerase chain reaction (PCR) products, the single nucleotide polymorphisms (SNPs) within TNF-? and TNF-? gene among 56 patients with chronic severe hepatitis B and 71 patients with chronic mild hepatitis B or asymptomatic carriers as well as 90 healthy controls were analyzed. The TNF-? concentration of serum in 56 patients with chronic severe hepatitis B and 30 healthy controls were determined by radio immunity assay (RIA). Results The frequencies of the TNF1/2 genotype and the TNF2 allele were significantly increased in patients with chronic severe hepatitis compared with healthy controls (25% vs 11.1%,P=0.028;12.5% vs 5.6%,P=0.036) and patients with chronic mild hepatitis B and asymptomatic carriers (25% vs 8.5%,P=0.011;12.5% vs 4.2%,P=0.015). Furthermore, heterozygotes carrying TNF2 allele had significantly higher levels of serum TNF-? than homozygotes for the wild type allele among all patients with chronic severe hepatitis B (P

Objective To investigate the ability of repairing bone defect with the compound of hydroxyapatite(HA),red bone morrow (RBM), bone morphogenic protein(BMP)and fibrin sealant (FS),and the feasibility with the compounds as bone substitute material. Methods The animal models of bilateral radius bone defect were created by surgery in the New Zealand white rabbits, and were treated with the compound of HA, RBM, BMP and FS, by autograft and no implant as control.The effect were observed by gross, histopathological and X-ray examinations, and were determined by biomechanics at 2,4,8 and 12 weeks after operation. Results By gross, histopathological and X-ray examinations, the effect indicated that the bone defect were perfectly repaired with autograft and the compound of HA, RBM, BMP and FS at 12 weeks, but not with the no implant group. The effect of biomechanics had no statistically significant difference between the autograft and the compound of HA, RBM, BMP and FS. Conclusion The compound of HA, RBM, BMP and FS possesses much high bone inductive potentialities and the ability of rebuilding bone defect and can serve as an autograft substitute material.

A prospective study was conducted for 91 cases of fever patients after neurosurgical procedures during the period of January-December 2013.They were divided into non-infection (n =42) and infection (n =49) groups according to whether there was infection complication.And another 51 nonsurgical hospitalized patients without fever or infection were selected as control group.The levels of procalcitonin (PCT),C-reactive protein (CRP) and white blood cell (WBC) were detected respectively.Statistical analysis showed that the level of PCT had no significant difference between non-infection and control groups (P ＞ 0.05).However,it was markedly elevated in infection group than non-infection group (P ＜ 0.01).The level of CRP was significantly different between non-infection and control groups (P ＜ 0.01).And it was the same between infection and non-infection groups.The level of WBC had significant difference between non-infection and control groups (P ＜ 0.05) and infection and non-infection groups (P ＜ 0.01).The receiver operating characteristic (ROC) curve showed that either sensitivity or specificity of PCT was the highest.Compared with CRP and WBC,PCT may identify more accurately the causes of fever after neurosurgical procedures.

Objective To discuss clinical results of treatment of trimalleolar fractures by minimally invasive surgical osteosynthesis. Methods From January 2002 to Doctober 2005, twenty-eight cases (mean age: 38.7 years) of trimalleolar fracture were treated by minimally invasive surgical osteosynthesis. Their lateral and posterior ankle joints were exposed through the Gatellier-Chastang incision. The sequence of reduction and fixation of ankle fracture was firstly the posterior ankle, then the medial and lateral malleolus, and distal tibiofibular syndesmosis lastly. Postoperatively, all the patients were fixed externally from 3 to 4 weeks with plaster splint. Results Follow-ups of 18 months on average revealed that all the cases healed. The healing time ranging from 2.8 to 4.5 months averaged 3.2 months. According to the Baird-Jackson scoring system, the results were rated as excellent in 16 cases, good in eight cases, moderate in three cases, and poor in one case, with the good-excellent rate being 85.7% . Conclusions The anatomical reduction and firm internal fixation are key factors in treatment of trimalleolar fractures. The minimally invasive surgical osteosynthesis is a good method due to the minimal invasion, a high rate of union, and fewer complications it results in.

Objective To evaluate the surgical effect of the surgical removal of both medial temporal lobe lesion and hippocampus amygdala for treating epilepsy. Methods Retrospectively analyzed 18 cases of epilepsy induced by the medial temporal lobe lesion and their hippocampal epileptic discharge was recorded by the deep electrode. Removed both medial temporal lobe lesion and hippocampus amygdala through medial temporal gyrus by modified pterional approach. The lesion had been totally removed in all of these 18 cases in naked eye. Evaluated the effect of surgery for epilepsy by Engel grading scale. Results These cases were followed up for average 2.8 years. Engel Ⅰ for 13 cases, Engel Ⅱ for 4 cases, Engel Ⅲ for 1 cases, Engel Ⅳ for none after operation. But there were lateral 1/4 quadrantanopsia in 2 cases, recent memory decreasing in 3 cases and none of death or any other complication. Conclusion Surgical removal of both medial temporal lobe lesion and hippocampus amygdala is a safe and effective method for treating epilepsy with less complication.

Objective By extra-body imitation release test with prolongation,medicines action on sustainedreleased injection of hydro-cortisone microeapsule was understood. Methods Making use of HPLC method was inspected to result of medicinal lower and die, solution. At the same time, the disaolvent course of microcapsule wall was observed by microscope and electron microscope. Results The release test at beginning 12 - 24 hours with rdease drugs measure was very lower-degree,along with the time more prolong in mierocapsule substance to contact of PBS solution(24～48h) was began disintegrate. The rdease of medicinal concentration also accompany with rising, the rising different concentration PBS solution from starting up to the highest as to general need 3～5 days and keep releasing for several days. Conclusion Sustained-released injection of hydrocortisone mierocapsule can be effectual action of prolongation released time in hind hydrocortisone.

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.

ObjectiveTo evaluate the neuroprotective effect of gradient perfusion-rewarming after deep hypothermia circulatory arrest (DHCA) in piglets.Methods12 Shanghai piglets (3-4 weeks old) were randomly divided into two groups of A (experiment group) and B (control group),average weight (9.78 ±0.93)kg.Animal CPB model is completed with microinvasive technique.DHCA duration is 90 min in two groups.During the rewarming period,group A was rewarmed with gradient perfusion strategy,maintain the temperature for 15 min every 5 ℃ elevation of the core temperature.Group B was rewarmed according normal consistent rewarming strategy.PH-stat management is adopt in both groups.Blood gas analysis,rectal temperature,heart rate,ECG,blood flow rate of carotid artery,glumatic acid/aspartate level of jugular vein and protein NFB of brain tissue are monitored during and/or after the cardiopulmonary bypass (CPB).ResultsDuration of rewarming in group A is (67.3 ± 7.8) min,and (41.8 ± 3.6)min in group B (P ＜ 0.05).Sample collected at the beginning of CPB,15 min of rewarming,30 min of rewarming and 45 min of rewarming show that there is no difference between the blood flow rate at 15 min of rewarming; difference are shown at the 30 min and 45 min of rewarming (P ＜ 0.5 ).High performance liquid chromatography ( HPLC ) analysis show the obvious difference of glumatic acid level of jugular vein at 30 min of rewarming and 45 min of rewarming ( P ＜ 0.5),this kind of difference of aspartate can only be seen at the 45 min of rewarming.Histologic evaluation shows gradient rewarming has a better effect on preservation of CA1 area neuron in hippocampus,however,Immunohistochemistry doesn't find the same effect.ConclusionControlled gradient perfusion-rewarming strategy can improve the neuroprotective effect during DHCA,keeping the balance of the blood flow,cerebral local temperature and brain metabolism might be the mechanism.

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.