Objective To optimize the most favorable extraction conditions of Nutmeg. Method With the volatile oils extracting ratio of Nutmeg as optimization targets, the extraction factors affected the volatile oils extracting ratio were optimized by uniform design. Result The optimum condition for extraction was as follows:medicinal materials which screen mesh were 4～10, being soaked in 14 times water for 1 hours and distilled for 6 hours with vapour. Conclusion The extraction process of optimization is simple and practicable, and can provide the test foundation for production.

In order to develop ELISA method for measuring the degradation fragment of fibronectin (MAD2),hybridoma technique was used to obtain the monoclonal antibody (McAb)IgG 1 against MAD2 without cross reaction with fibronectin.The sera from 277 patients with hepatocellular carcinoma (HCC),76 with metastatic hepatic carcinoma (HMC),98 with alimentary canal carcinoma (ACC) and 156 with chronic liver disease(CLD) and 48 healthy subjects were assayed with ELISA method using this antibody.The examination showed that the mean value of MAD2 from the patients with HCC showed obviously significant difference compared with those of CLD,HMC,ACC and normal control groups ( p

OBJECTIVE To observe the efficacy of lemon eucalyptus oil on disinfection of indoor air.METHODS Fumigating method and air sampling method were used to evaluate the disinfection efficacy.Ultraviolet disinfection method was used as control.RESULTS Aerosol spraying disinfection with the lemon eucalyptus oil at a dosage of 10 ml/m3 followed by closing the room for 30 min can killed 95.4% of the natural bacteria of indoor air.Irradiation by 30 W ultraviolet lamp for 30 min under same conditions killed 89.7% of the natural bacteria of indoor air.CONCLUSIONS The efficacy of the lemon eucalyptus oil disinfectant at 10ml/m3 concentration in killing natural bacteria in indoor air fulfils the requirement of eligible disinfection and is better than the disinfection efficacy of ultraviolet irradiation.

Objective To investigate the inhibitory effect of hydroxy safflor yellow A ( HSYA ) on angiogenesis of H22 tumor-bearing mice and it's effects on the protein expression of MMP-3 .Methods After establishing the hep-atoma model for 24 h, the mice were randomly divided into control group , sorafenib group and HSYA group , the dose HSYA group received intraperitoneal injection at different dosages (1.125 and 2.25 mg/kg).The pathologi-cal changes were examined with HE staining , immunohistochemical staining and Western blot were applied to meas-ure the expression of angiogenesis related factor ( MMP-3 ) and we also detected the microvessel density with CD 34 . Results Compared with control group , the tumor cells proliferation and the new angiogenesis in HSYA group were suppressed .The expression of MMP-3 in HSYA group was significant reduced .Especially the dose of 2.25 mg/kg HSYA group ( P<0.01 ) , and tumor MVD-CD34 was also significantly reduced ( P<0.01 ) .But the effect is not better than sorafenib group .Conclusions HSYA may inhibit angiogenesis of tumor tissue in a certain concentration range and the anti-angiogenesis effect of HSYA may be related to inhibition of the protein expression of matrix met-alloproteinase-3 .

Objective: To study the construction and application of a vector with Cre recombinase recognition site lox66 for mouse HPRT gene targeting in embryonic stem(ES) cells. Methods: Using the HPRT genomic DNA fragment and synthesized oligonucleotides, pSP HPRT lox66 Neo was designed and constructed as a replacement vector by common molecular cloning techniques. After the linearized pSP HPRT lox66 Neo DNA was electroporated into ES cells, and transfected cells were cultured in G418/6 TG drug selection medium. The recombination efficiency of this vector was tested. Results: The main components of pSP HPRT lox66 Neo were a positive selection gene Neo, lox66, long and short homologous fragments of mouse HPRT gene and plasmid backbone. Twenty ES cell clones with HPRT gene inactivated were obtained. Conclusion: An effective replacement vector with Cre recombinase recognition site lox66 is constructed and applied to HPRT gene targeting in ES cells.

Objective To sum up the experience for the treatment of splenic trauma. Methods Data of 129 cases of traumatic splenic rupture from 1984 to 2004 were retrospectively analyzed. Results Conservative management was undertaken in 17 cases, splenorrhaphy in 16 cases, total splenectomy in 86 cases, partial splenectomy in 8 cases, and splenic autotransplantation in 23 cases. The results were all satisfactory. Conclusion In splenic trauma, treatment modality should be on adopted case to case basis according to the types of splenic rupture. Combined splenic salvage can be used on certain conditions.

ObjectiveTo investigate the correlation of MRI features and phenotypes and genetic mutations in Pelizaeus-Merzbacher disease.Methods Sixteen boys with clinical diagnosis of PelizaeusMerzbacher disease (PMD) were included in this study.Their ages ranged from 22 months to 9 years.They were examined by pediatric neurologists,and clinical classification was made according to the symptoms and physical signs.An experienced radiologist reviewed the cranial MRI images and analyzed the brain involvement,including pallidus globus,pyramidal tract,corpus callosum,cerebellar white matter,semiovale centrum,brain atrophy and ‘ tigroid sign’.ResultsThere were 8 patients with classic form,7 patients with transitional form and one patient with connatal form.They all showed diffuse delayed myelination in the white matter,with involvement of pallidus globus in 13 cases,pyramidal tract in 7 cases,corpus callosum in 11 cases,cerebellar white matter in 7 cases,semiovale centrum in 12 cases.Cerebral atrophy was found in 5 patients and eerebellar atrophy was found in one patient.Five cases depicted ' tigroid sign'.In patients with PLP1 gene point mutation,pyramidal tract and cerebellar white matter involvement showed a high incidence.Cerebellar white matter lesions were relatively frequent in children with transitional form and connatal form.In contrast,‘ tigroid sign' was often related to classic form,which indicated a better myelination and outcome.ConclusionPMD patients show distinct imaging features in their brains,which may be correlated with the phenotype and genetic mutation.

BACKGROUND:To construct tissue engineering cartilage would open up a novel way for the repair of cartilage damage in avoidance of the disadvantages of traditional therapeutic method.OBJECTIVE: To probe the techniques for the isolation of mesenchymal stem cells (MSCs) from bone marrow, as well as the in vitro differentiation into chondrocytic phenotype in a specific culture fluid.DESIGN:A complete randomized experimentSETTING:The Department of Traumatic Orthopedics and Hand Surgery of the First Affiliated Hospital of Guangxi Medical University, and Teaching and Research Faculty of Histology and Embryology of Guangxi Medical University.METHODS: The experiment was carried out at Guangxi Medical University between August 2002 and April 2003. Twenty SD neonatal weaning rats were selected. Bone marrow was aspirated from the bones of rat limbs and was isolated by gradient centrifugation in Percoll, and MSCs could be obtained in combination with adherent screening method, which were then cultured in DMEM-LG with 15% fatal bovine serum (FBS) in the incubator of 37℃ with 5% CO2 for 10-14 days. The passage cells were induced in DMEM-HG with 15% FBS (containing TGF-β1 10 μg/L, 10-7 mol/L dexamethasone, 50 mg/L VitC).MAIN OUTCOME MEASURES :The morphology, growth, as well as proliferation and specific expression of chondrogenic matrix of in vitro cultured MSCs due to specific induction.RESULTS: Totally 20 SD rats were observed and analyzed with no loss SCs grew in visible symmetric colonies, displaying a long-spindle shape,and the morphological characteristics of marrow-derived MSCs had no obvious changes during passage-culture, but its proliferation time was found from a shuttle fibroblastic appearance to polygonal shape, displaying posiHC staining of type Ⅱ collagen of cartilage specific matrix.bronectin adherent screening technique is a convenient, effective and practical method to separate and collect MSCs from rat bone marrows in chondrogenic phenotype when induced by a specific medium and can secrete cartilage specific matrix, and they can be the optimal seed cells for cartilage tissue engineering.

Objective To investigate the changes of nitric oxide content and inducible nitric oxide synthase (iNOS) activity in rat brain tissues after explosive craniocerebral wound and observe the influence of emodin on these changes. Methods After rats were subjected to electric detonators, emodin (10 mg/kg) was injected intraperitoneally at 10 minutes. Water content of brain tissues was measured by using dry-wet methods. The content of nitric oxide and the activity of iNOS were measured at 2, 4, 12 and 24 hours respectively after wound by means of nitrade reductase and colorimctrie method. Results The content of nitric oxide and the activity of iNOS kept at a high level (P<0.01). The content of nitric oxide and the activity of iNOS in model group were increased significantly and reached peak at 2 hours after wound. While the content of nitric oxide and the activity of iNOS in emodin group were significantly lower than that in model group (P<0.01). Conclusion Emodin plays a role in decreasing water content of brain tissues and inhibiting activity of iNOS and may exert a protective effect on brain injury during pathological course after explosive craniocerebral wound.

Objective To investigate the risk factors for early hypoglycemia and its relationship with prognosis of patients with cerebral infarction.Methods A total of 273 patients with cerebral infarction were divided into the normal blood glucose(NBG) and severe hypoglycemia (SHG)and mild hypoglycemia(MHG) groups in our hospital.Biochemical indicators,the National Institute of Health stroke scale(NIHSS)and mortality were compared between the three groups.According to prognosis,patients were divided into death group and survival group.The NIHSS score,blood glucose concentration and incidence of hypoglycemia were compared between death and survival groups.Pearson relationship between hypoglycemia and NIHSS score,and spearman rank correlation between hypoglycemia severity and mortality were analyzed.Results Levels of lactic acid (6.3 ± 2.8) mmol/L,creatinine(268.7 ± 63.9) mmol/L,urea nitrogen (13.8 ± 3.7) mmol/L,albumin (25.6 ±4.9) g/L,alanine aminotransferase (150 ± 19.7) U/L,NIHSS (22.3 ± 9.2) scores,and mortality rates (38.1 ％)were higher in severe hypoglycemia group than in both NBG group and severe hypoglycemia group[(lactic acid:4.7±2.3 mmol/L and 3.3±1.5 mmol/L),(creatinine 134.8±51.3 mmol/L and 78.7±40.8 mmol/L),(urea nitrogen 7.9±4.2 mmol/L and 7.7±3.3 mmol/L),(albumin 36.9±3.8 g/L and 35.6±4.3 g/L),(alanine aminotransferase 85.8± 18.3U/L and 46.3± 13.8U/L),(NHISS 14.6±5.9 scores and 10.5 ± 5.4 scores)and(mortality rates 20.8％,11.0％)] (all P＜0.01).There was a negative correlation between hypoglycemia and NIHSS score(r=-0.45,P＜＜0.05).There was a positive correlation between hypoglycemic severity and mortality (r =0.41,P ＜ 0.05).Multiple Logistic regression showed that creatinine and alanine aminotransferase were correlated with hypoglycemia and prognosis of patients with cerebral infarction(both P＜0.05).Conclusions Early hypoglycemia in patients with severe cerebral infarction is closely correlated with the liver and kidney insufficiency,and a severe cerebral infarction combined with hypoglycemia often indicate a poor prognosis.