Objective To study the expression of VEGF, MMP-2 and MMP-9 in benign and malignant lesions of breast and their effects on breast carcinoma's invasiveness and metastasis. Methods VEGF,MMP-2 and MMP-9 expression was detected by SP method in 20 breast fibroadenoma and 12 normal breast specimens taken from non-cancerous regions adjacent to breast cancer tissue and 38 breast carcinomas. Results The expression rate of VEGF, MMP-2 and MMP-9 in breast carcinoma was remarkably higher than those in normal breast tissue and breast fibrogdenoma (63.2% vs 25.0% and 30. 0% ,78. 95% vs 33.3% and 35.0% and 71.1% vs 41.7% and 45.0% ,respectively; P ＜0.05). There was a correlation between VEGF, MMP-2 and MMP-9 expression and nodal metastasis ( P ＜0.05). High expression of VEGF, MMP-2 and MMP-9 was correlated with pathological grade ( P ＜ 0.05, respectively). The overexpression of VEGF was related with those of MMP-2 and MMP-9 in breast cancer( VEGF and MMP-2:rs =0.541, P ＜0.01 ;VEGF and M M P-9: rs = 0.11, P ＜ 0.01; MMP-2and M MP-9: rs= 0P ＜ 0.01 ). Conclusion VEGF,MMP-2 and MMP-9 overexpression is strongly related to the invasiveness and metastasis of breast carcinoma, which might be valuable in evaluating tumor invasion and metastasis.

Objective To observe the Clinical effects on Sanao tables combined with Montelukast Tables for cough variant asthma (CVA) (cold beam lung card).Methods Eighty-nine cases of sydrome differentiation (cold beam lung card) complied with CVA diagnostic criteria were randomly divided into treatment group (45 cases,3 cases were lost) and control group(44 cases,4 cases were lost)).The treatment group was given oral San'ao tables combined with Montelukast tables,and the control group was given oral Montelukast tables.The course of treatment were all 14 days.Chinese medical syndrome,cardinal symptom and airway reactivity effects were determined and compared at pretherapy.Results Clinical effects of Chinese medical syndrome (effective rate (90.48％,38/42),cough severity (85.71％,36/42),frequency (90.48％,38/42),cough up phlegm (85.71％,36/42),pharyngeal itching (80.95％,4/42) and aversion to cold (76.19％,32/ 42)) in treatment group were all superior to control group (70.00％ (28/40),62.50％ (25/40),70.00％ (28/ 40),55.00％ (22/40),52.50％ (21/40),25.00％ (10/40)).The differences were statistical significance (x2=14.046,14.046,13.923,23.104,17.729,17.729;P＜0.05 or P＜0.01).Conclusion Clinical effects of San' ao tables combined with Montelukast tables on CVA is superior to Montelukast tables.

[Objective]To investigate the clinical application of treating soft tissue defects of the feet、ankle and lower one third of the legs with reverse saphenous neurocutaneous concomitant vessel pedicled island flap.[Method]From March 1999 to June 2003,six cases of soft tissue defects of the feet,ankle and lower one third of the legs were treated with reverse saphenous neurocutaneous concomitant vessel pedicled island flap.The size of flaps were from 12 cm?7 cm to 5 cm?3 cm,aver aging 8 cm?5 cm.[Result]Flaps of 6 cases survived all showing good quality.After 10 months to 3 years follow-up,all of the six cases were thinked very good.[Conclusion]The reversed saphenous neurocutaneous concomitant vessel pedicled island flap treating soft tissue defects of the feet,the ankle and the lower one third of legs is easy to be performed.The flap has liable blood supply and does not sacrifice the major arteries.

Objective The microcapsules were prepared by using chitosan and sodium alginate as wall materials,and the controlled-release microcapsulas of interferon tau were hoped to be develop an oral interferon preparation.Methods The microcapsules were prepared by using syringe hand-made drop.In the process of dropping,speed and distance were the major factors which influence the form of microcapsules.Results Chitosanalginate microcapsules which was prepared simple and fast had high encapsulation rate and extended-release effect in intestine.Conclusion Chitosan-alginate microcapsules have the potential to be used to prepare interferon tau and other protein medicines.

Objective To study the optimum procedure of extracting ginsenoside from Panax ginseng by super pressure extracting technique (SPET). Methods SPET was used to extract ginsenoside from P. ginseng at the normal temperature. The optimum extracting process was selected by the orthogonal test. The contents of ginsenoside in P. ginseng were determined by UV-spectrophotometry. Results The optimum condition of SPET was as follows: when the solid dissolved in 50% ethanol, ratio of raw P. ginseng (g) and solvent (mL) was 1∶75, the pressure was kept at 500 MPa for 2 min; the high yield of ginsenoside was up to 7.76%. Conclusion SPET has a series of advantages, such as higher efficiency, shorter extracting time, and lower exhausting energy, at the same time, the impurity is little, etc.

Objective: To study the effect of intraarticular injection of crosslinked-chitosan in the treatment of knee osteoarthritis in rabbits. Methods: Thirty-two New Zealand white rabbits were randomly divided into 4 groups (groups A, B, C, and D; 8 rabbits in each group). The knee osteoarthritis models were prepared by anterior cruciate ligament transection in the left hind in groups A, B, and C. At 4 weeks after operation, the rabbits were received intraarticular injection of 0.6 mL crosslinked-chitosan in group A, 0.3 mL chitosan (once per 2 weeks, for twice) in group B, and 0.3 mL saline (once per 2 weeks, for twice) in group C. The rabbits in group D were treated with sham operation in the left hind, and received intraarticular injection of 0.3 mL saline (once per 2 weeks, for twice). At 8 weeks, the macroscopic observation, histological examination (HE staining, Safranin-fast green double staining, and Mankin score), scanning electron microscopy (SEM) observation, and immunohistochemical staining of collagen type Ⅱ were performed. Results: Macroscopic and SEM observations showed that the cartilage in group D was basically the same as normal and better than that in groups A and B, and the abrasion of cartilage in group C was the most serious. The histological observation results in groups A and B were slightly similar and better than those in group C, but not up to the structure of group D. The macroscopic score and Mankin score of groups B and C were significantly higher than those of group D ( P<0.05), and there was no significant difference between group A and group B ( P>0.05). Immunohistochemical staining results showed that the collagen type Ⅱ positive percentage of chondrocytes was significantly higher in group D than that in groups B and C, and no significant difference was found between group A and group B ( P>0.05). Conclusion: The crosslinked-chitosan can significantly improve the osteoarthritis of the rabbit knee, delay the pathological changes of osteoarthritis, and decrease the frequency of injection.

Objective To investigate the relationship between the secondary hypertension and pituitary microadenoma,and to explore the diagnostic value of magnetic resonance imaging(MRI) examination for pituitary microadenoma.Methods We analyzed the clinical information,laboratory examination results and imaging data in 21 hypertension patients complicated with pituitary microadenoma.Results The manifestations of the 21 patients were as follows:(1) The onset of hypertension was in young age,complicated with headache;(2) The results of 24-hour ambulatory blood pressure monitor(24h ABPM) showed the disappearance of circadian rhythm of blood pressure,and obvious increase of systolic/diastolic blood pressure;(3) The antihypertensive effect of antihypertensive drugs was not satisfactory;(4) Of adenohypophyseal hormones,adrenocorticotrophic hormone(ACTH) level in most patients and prolactin(PRL) level in a few patients were higher than the normal level,and the other hormones levels were normal;(5) The results of MRI examination presented adrenal cortical hyperplasia,adrenal adenoma and chromaffin tumor in some patients.(6) The results of MRI examination showed pituitary microadenoma in all of the patients.Conclusion For those middle-aged and young hypertension patients on whom antihypertensive effect of drugs is poor,MRI examination for adrenal gland and pituitary gland should be taken as the routine examination.MRI examination is the optimal imaging method for pituitary microadenoma,and supply evidence for the syndrome differentiation and treatment of pituitary microadenoma.

AIM: Site-specific functional neurons of brains were with different cellular morphology. It has not been fully understood whether the grafted neural stem cells could differentiate into the site-specific neurons. This experiment is to investigate the neuronal differentiation of the neural stem cells derived from a human fetal brain after transplanted into young rats' brains, to study the possibility of cell-replacement therapy for children's brain disorders with neural stem cells. METHODS: Experiments were performed at the Cell Laboratory of Naval General Hospital from April to July 2007. ①Human fetal brain tissues of 16 week gestation were provided by Department of Gynaecology and Obstetrics of Naval Hospital. Pregnant woman and family members signed an informed consent. Experimental intervention was approved by Hospital Ethical Committee. Fourteen clean brood young SD rats aged 10 days, irrespective of gender, were provided by Experimental Animal Center of Medical College of Peking University. Animal intervention met the animal ethical standards. ②The neural stem cell spheres were derived from the fetal brain tissues of 16 week gestation. The differentiation multipotency of the neurosphere was identified when cultured in a child's cerebrospinal fluid (CSF). The neurospheres cultured in vitro for 14 days were injected into the lateral ventricles of young rats of 10 days old. The rats were respectively killed at days 4, 7 and 14 after transplantation. The special immuno-fluorescent assays were performed using anti-human neurofilament (anti-hNF) to show the location and morphology of graft neurons. RESULTS: ①The typical floating neurospheres were obtained, with the potency to differentiate into neurons, astrocytes and oligodendrocytes. ②The neuronal differentiation of grafts was detected with the mixture of three monoclonal antibodies against human neurofilament. Four days after transplantation, the immune response positive cells lied within the granule cell layer of cerebral cortex were shown in the shape of granule cells, or within the pyramid cell layer in the shape of pyramid cells with long processes, and the interneuron-like cells also were seen. The Purkinje cells arranging in a monolayer were detected in the cerebellum. Compared the results at different time points, the location of grafts were the same. The graft cells were less and the processes were longer over time. CONCLUSION: The in vitro cultured neurosphere cells can migrate into brain tissues and differentiate into site-specific neurons in shape after transplanting into the lateral ventricles of young rats. It is suggested that the host brain tissue microenvironment played an important role in guiding the graft differentiation into neurons. The results have an important significance for understanding cell replacement of developing brain disorders.

Objective To detect apoptosis of transplanted lung adenocarcinoma cells in nude mice after 125I brachytherapy by 99Tcm-Annexin V combined with diffusion weighted MRI (MR-DWI).Methods Twenty-five BALB/c-nu nude mice models subcutaneously transplanted with A549 cells were divided into experimental group (EG,n=13) and control group (CG,n=12) by random number table method.One 125I seed with apparent activity of (24.8±6.3) MBq was implanted into each mouse in EG,while CG underwent cold seed implantation.Both of 99Tcm-Annexin V imaging and MR-DWI were performed within 7-10 d after brachytherapy,then all mice were sacrificed and tumor cell apoptosis was detected by terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) immunofluorescence,Survivin expression was assayed by SP.Two-sample t test,x2 test and Pearson correlation analysis were used for data analysis.Results Positive rate of cell apoptosis by 99Tcm-Annexin V imaging was 69.2％(9/13) and 8.3％(1/12) respectively in EG and CG (x2 =12.73,P＜0.01).The uptake ratio of 99Tcm-Annexin V,apparent diffusion coefficient(ADC) value and apoptosis index(M) of the tumor in EG were 2.91±0.85,(2.03±0.44)×10-3 mm2/s and (49± 18) ％,respectively,which were significantly higher than those of CG (1.26 ± 0.37,(1.29 ± 0.21) ×10-3 mm2/s and (11±4)％ respectively,t=5.930,5.452,7.606,all P＜0.05).Survivin expression in EG and CG was (46± 13) ％ and (15±7) ％ respectively (t =5.158,P＜0.05).The value of ADC was correlated with AI and uptake ratio(r=0.756,0.788,both P＜0.05).Uptake ratio was correlated with AI (r=0.754,P＜0.05),while Survivin expression was negatively correlated with AI (r =-0.772,P＜0.05).Conclusions Down-regulation of Survivin expression may play an important role in apoptosis induced by 125I brachytherapy.99Tcm-Annexin V combined with MR-DWI could effectively evaluate apoptosis of lung adenocarcinoma cells in a non-invasive way,thus it might be helpful in evaluation of early efficacy of 125I brachytherapy.

Objective To study the relationship between the mutation of the inverted repeat (IR) gene in the multiple transferable resistant (mtr) system and multiple antibiotic resistance of Neisseria gonorrhoeae. Methods The antimicrobial susceptibilities of isolated strains were tested. An agar plate dilution method was used to determine the minimum inhibitory concentrations. The target genes were amplified by PCR and subjected to sequencing. Results No mutation was found in the IR gene of either of 2 sensitive or 5 penicillin-resistant Neisseria gonorrhoeas strains. Among the 17 multiple-antibiotic-resistant strains, a strain with both azithromycin- and penicillin-resistance had T/A and T/A insertions, and another had A/T deletion. Conclusion Mutations in the IR gene of the mtr system of Neisseria gonorrhoeae might result in multiple antibiotic resistance.