Objective To investigate the relationship between plasma cell mastitis and prolactin. Methods The relationship between the lesion area , degree of inverted nipple and prolactin level of 228 plasma cell mastitis patients were observed , as well as their prolactin level before and after treatment , to explore the factors that influence relapse. Results There was no significant statistical relationship between prolactin level and lesions area and degree of inverted nipple. Prolactin level before and after treatment were statistically different (t =2.347,P = 0.02). Menstrual status, nipple status, comorbidities, lesion area and prolactin level were considered, only prolactin level was related with disease recurrence (P = 0.038). Conclusion prolactin level could significantly reduced as the disease cured , but elevated prolactin may lead to relapse of the disease.

Objective To compare clinical outcomes of double-bundle and single-bundle in individualized arthroscopic anatomical reconstruction of anterior cruciate ligament (ACL) . Methods The clinical data of 117 patients were reviewed who had received double-bundle or single-bundle arthroscopic ACL reconstruction from March 2007 through September 2009 in our hospital and had undergone complete follow-up. Of them, 35 cases had single-bundle ACL reconstruction and 82 double-bundle reconstruction. In the single-bundle group(group A), there were 31 men and 4 women, aged 28. 6 ±5. 1 years. In the double-bundle group(group B), there were 73 men and 9 women, aged 27. 6 ±5. 4 years. The 2 groups were comparable in the preoperative demographic data ( P > 0. 05). To evaluate the outcomes, Lachman and Pivot Shift exams , KT-2000, Lysholm and IKDC (International Knee Documentation Committee) scores, were adopted. Results The 117 patients received a mean follow-up of 15 months (from 11 to 25 months). The Lachman test showed 88. 6% (31/35) were normal in group A and 95. 1% (78/82) were normal in group B.The pivot-shift test showed 88. 6%(31/35) were normal in group A and 96. 3% (79/82) were normal in group B. Group A had a mean Lysholm score of 93. 4 ± 8. 2 and group B a mean Lysholm score of 93. 7 ±7. 0. There were no significant differences between the 2 groups in the above indexes ( P > 0. 05). By IKDC score, 71. 4% (25/135) were normal in group A and 93. 9% (77/82) were normal in group B. The KT-2000 test showed a mean of 1. 4 ± 0. 6 mm in group A and a mean of 1. 1 ± 0. 5 mm in group B. These 2 values were significantly different between the 2 groups ( P < 0. 05). Conclusions The individualized arthroscopic double-bundle anatomical reconstruction of ACL can maximally restore the anteroposterior and rotational stability. Arrangement of the ACL insertion site on the femoral and tibial side, three-portal technique and ruler application are keys for individualized anatomical double-bundle ACL reconstruction.

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

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.

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: 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 study the mechanism of 5 fluorouracil(5 FU) in the treatment of severe acute pancreatitis(SAP). Methods SD rats were randomly divided into three groups:acute pancreatitis group (AP group,n=12), 5 FU treatment group( 5 FU group,n=10),and control group (n=10). In 5 FU group,5 FU(1mg/100g) were injected immediately after AP was induced.12h after the AP was induced, blood samples were taken to determine nitric oxide(NO) and interleukin 6(IL 6). Results Compared with AP group,in 5 FU group, the survival rate increased,mean survival time prolonged,ascites volume decreased significantly,and the inflammatory mediators subsided. Conclusions NO and IL 6 may play an important role in the pathogenises of acute pancreatitis. The treatment effect of 5 FU for AP may through inhibit the inflammatory mediators.

Breast cancer is called "Ruyan" in literature of traditional Chinese medicine. We synthesized the ancient and contemporary discussions and raised the theory that "Duxie" (poisonous pathogenic factor) is the etiological factor and pathologic product through the whole course of breast cancer. "Liuyin Fudu" (latent poison of six exogenous pathogenic factors) and "Qiqing Yudu" (stagnant poison of seven emotions) are the main etiological factors affecting the breast cancer occurrence. "Aidu Neisheng" (internal product of cancer poison) is the essential change in breast cancer occurrence. "Tandu Yujie" (stagnation of phlegm, poison and blood stasis) is the essential pathogenesis of the breast cancer's development. "Yudu Weiqing" (vestigial poison) is the main pathogenesis of breast cancer after operation. "Yudu Pangcuan" (vestigial poison invasion elsewhere) is the key pathogenesis of recurrence and metastasis after operation. "Sanjie Jiedu" (dispersing accumulation and detoxification) is an important therapeutic principle in breast cancer's treatment after operation. The "Tandu Yujie" pathogenesis theory and "Sanjie Jiedu" therapeutic principle developed the theory about breast cancer in traditional Chinese medicine, and have some clinical application value.

OBJECTIVE: To observe the effect of Ruyiping, a traditional Chinese compound herbal medicine composed of 5 Chinese herbs for removing toxic materials and dissipating nodules from Runing II, another traditional Chinese compound herbal medicine for treating breast cancer, in preventing recidivation and metastasis in breast cancer patients after operation. METHODS: Eighty patients with breast cancer after operation were randomly divided into Ruyiping group and Runing II group, and prescribed Ruyiping and Runing II on the basis of chemotherapy, radiotherapy and endocrine therapy respectively for two years. RESULTS: There were two patients with metastasis and three patients lost to follow-up in Ruyiping group and three and two in Runing II group. The recidivation and metastasis rates were 5.41% and 7.89% respectively. The difference between the two groups was not statistically significant (P>0.05). The difference of disease-free survival time between the two groups was also not statistically significant. CONCLUSIONS: The effect of Ruyiping in preventing recidivation and metastasis is similar to that of Runing II. Ruyiping is the essential component of Runing II for preventing recidivation and metastasis. The result provides some clinical evidences for the theory that "Yudu Pangcuan" (vestigial poison invasion elsewhere) is the essential pathogenesis of breast cancer's recidivation and metastasis and the utilization of "Sanjie Jiedu" (dispersing accumulation and detoxification) is the therapeutic principle in preventing recidivation and metastasis after operation.