Objective To establish a RP-HPLC method investigate the processing technique and mechanism of Eucommiae Cortex.Methods The RP-HPLC method was applied to simultaneously determining six ingredients,geniposidic acid,geniposide,genipin,chlorogenic acid,(+)-pinoresinol-di-β-D-glucopyranoside,and(+)-syringaresinol-di-β-D-glucopyranoside,in the different processed barks of Eucommia ulmoides.Results The valid method with good accuracy could be well used to study the processing technique of E.ulmoides;Besides,target ingredients in E.ulmoide were decreased within 6 h when they were processed.Conclusion Established RP-HPLC is a reliable method which could be used to research the processing technique of the barks ofE.ulmoides.Moreover,the result of this study could be provided with significant evidence of processed barks of E.ulmoides.

Objective:To investigate the competency of informed consent in new admitted schizophrenia pa-tients.Methods:Fifty-three patients who were consecutively admitted to the hospital and diagnosed as schizophrenia according to the International Statistical Classification of Diseases and Related Health Problems,Tenth Revision (ICD-10)were recruited and assessed in seventy-two hours.The competence of informed consent was assessed by the MacArthur Competence Assessment Tool-Treatment (MacCAT-T).The insight was assessed by the Insight and Treatment Attitude Questionnaire.The severity of illness was assessed by the Brief Psychiatric Rating Scale (BPRS).Correlations between the competence of informed consent and the factors mentioned above were discussed.Results:Forty-five (84.9%)patients had lower MacCAT-T scores,especially the scores of understand-ing dimension (1.58 ±1.85)and appreciation dimension (2.40 ±1.47).The competence of informed consent was significantly correlated with insight (r =0.73,P <0.01)but not correlated with age,education,duration of illness and BPRS scores.Logistic regression demonstrated that insight (OR =0.69)was a protective factor of competence of informed consent while thought disturbance (OR =1.39)was a risk factor.As to the involuntary admitted pa-tients,34.1% of them had competence of informed consent.Conclusion:The competence of informed consent in new admitted schizophrenia patients is widely impaired especially in understanding and appreciation dimension.The competence of informed consent is correlated with insight and thought disturbance.Voluntary or involuntary hospi-talization does not reflect the level of competence of informed consent.

Objective The expression and neuroprotective effect of meteorin in neurons and astrocytes after cerebral infarc-tion have yet to be clarified.This study was to investigate the expression and location of meteorin in the rat model of middle cerebral ar -tery occlusion (MCAO) and its neuroprotective effect against oxygen -glucose deprivation (OGD)-induced injury in cultured neurons or astrocytes. Methods Forty-two healthy adult male SD rats were randomly divided into 6 groups of equal number:sham operation and 6 h, 12 h, 24 h, 3 d, and 7 d MCAO.The cortical tissue was harvested for determination of the expression and location of meteorin by Western blot and immunohistochemistry as well as the meteorin expression in the neurons and astrocytes subjected to OGD. The neuroprotective effect of meteorin on the neurons and astrocytes was e-valuated by CCK8 and PI/Hoechst33342 staining. Results Com-pared with the sham operation group, the expression of meteorin was decreased after MCAO and reached the lowest level at 3 days ( P<0.01) , mainly in the neurons and astrocytes.Exogenous meteorin helped the survival of the astrocytes subjected to OGD, with the A value decreased in the groups of OGD ( 0.63 ) , OGD +meteorin
(0.78), and OGD+PBS (0.60) as compared with the sham control ( 1.51) (P<0.01), with no statistically significant differences among the former three groups (P>0.05).Concerning the effect of meteorin intervention on the OGD-induced injury of the astrocytes, the A value was significantly reduced in OGD (1.24 ±0.17), OGD+meteorin (1.51 ±0.30), and OGD+PBS (1.23 ±0.16) in comparison with the sham control (2.43 ±0.12) (P<0.01), lower in the OGD and OGD+PBS groups than in the OGD+meteorin group ( P<0.05) . Conclusion Meteorin is mainly expressed in the neurons and astrocytes after MCAO and it promotes the survival of the astrocytes with OGD-induced injury.

BACKGROUND: Human keratinocyte growth factor-2 (hKGF-2) has extensive physiological functions, which plays an important role in embryonic development, tissue-repairing, nervous regeneration, vascularization and development of tumor.OBJECTIVE: To clone hKGF-2 gene, obtain the expression of hKGF-2 in Escherichia coli(E.coli) and determine its bioactivity, so as to provide experimental basis for further investigation.DESIGN: Open experiment.SETTING: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention.MATERIALS: The experiment was conducted in State Key Laboratory of Viral Genetic Engineering, Institute for Viral Disease Control and Prevention of Chinese Center for Disease Control and Prevention. The temperature control expression vector pBV220 was constructed by State Key Laboratory of Viral Genetic Engineering; EcoR Ⅰ , BamH Ⅰ , T4 DNA ligase (Promega Co., Ltd.); The specific polymerase chain reaction (PCR) of hKGF-2 (Manufactured by Shanghai Boya Biotechnology Co., Ltd.); Heparin-Sepharose CL-6B (Pharmacia Company); PCR rapid purification kit,Trizol kits for total RNA extract, Kits for RT-PCR (GIBCO Co., Ltd.); Kits for rapid extraction of plasmid DNA (Boda Company); BL-21-codon plus compent cells (Stratagene Co., Ltd.).METHODS: High-expression strain BL-21 codon plus competent cells was used to express and purify initially recombinant hKGF-2 protein, and its activity was detected. RT-PCR was adopted to obtain hKGF-2 cDNA from lung tissues of naturally aborted fetus and clone it into pBV220 carri er plasmid. The hKGF-2 protein expressed in BL-21 codon plus competent cells of E.coli. Affinity chromatography and ion exchange chromatography were applied in isolation and purification, and the bioactivity of expression protein was determined in cell proliferation test.MAIN OUTCOME MEASURES: The length and sequence of cDNA segment in hKGF-2, the expression of hKGF-2 gene inE.coli and the purification of hKGF-2 activity.RESULTS: The segment of hKGF-2 cNDA was about 500 bp, and hKGF-2 protein highly expressed in BL-21, which had soluble expression in the supernatant. SDS-PAGE showed that the relative molecular mass was about 20000, and hKGF-2 protein could significantly promote the mitotic activity of NIH3T3 cells. The A value (490 nm) of hKGF-2 in the 1 μg/L, 5 μg/L and 10 μg/L groupswere higher than that in the blank control group, and the differences were significant (which were 0.174±0.022,0.220±0.029,0.306±0.050,0.066±0.004 respectively,P ＜ 0.001).CONCLUSION: hKGF-2 gene is successfully cloned, which highly expresses in BL-21 of the E.coli. Purified hKGF-2 protein can stimulate the proliferation of NIH3T3 cells and significantly promote its mitotic activity.

Objective:To establish an FPLC-MS/MS method for the determination of fluvoxamine in human plasma. Methods:The separation was performed on an Inertsil? ODS-SP column(2. 1 × 100 mm, 3 μm). The mobile phase was acetonitrile-2 mmol· L-1 ammonium acetate (45: 55, v/v) containing 0. 1% formic acid at a flow rate of 0. 3 ml·min-1 . Lansoprazole was used as the internal standard( IS) . Electrospray ionization ( ESI) source was applied and operated in a positive ion model. Multiple reaction moni-toring (MRM) model with the transitions of fluvoxamine m/z 319. 1→m/z 69. 8 and lansoprazole m/z 370. 2→m/z 252. 1 was used to quantify fluvoxamine and IS, respectively. Results:In human plasma, the standard curve was linear within the range of 1-100 μg· L-1 . The lower limit of quantification of fluvoxamine( LLOQ) was 1μg·L-1 . The intra-day RSD was less than 5%, the inter-day RSD was less than 10%, and the method recovery was 85%-95%. Conclusion:The method is simple, sensitive, accurate and reproduci-ble. It is applicable in the pharmacokinetic study of fluvoxamine for clinical pharmacokinetics and bioequivalence studies.

Objective:To establish an FPLC-MS/MS method for the determination of fluvoxamine in human plasma. Methods:The separation was performed on an Inertsil? ODS-SP column(2. 1 × 100 mm, 3 μm). The mobile phase was acetonitrile-2 mmol· L-1 ammonium acetate (45: 55, v/v) containing 0. 1% formic acid at a flow rate of 0. 3 ml·min-1 . Lansoprazole was used as the internal standard( IS) . Electrospray ionization ( ESI) source was applied and operated in a positive ion model. Multiple reaction moni-toring (MRM) model with the transitions of fluvoxamine m/z 319. 1→m/z 69. 8 and lansoprazole m/z 370. 2→m/z 252. 1 was used to quantify fluvoxamine and IS, respectively. Results:In human plasma, the standard curve was linear within the range of 1-100 μg· L-1 . The lower limit of quantification of fluvoxamine( LLOQ) was 1μg·L-1 . The intra-day RSD was less than 5%, the inter-day RSD was less than 10%, and the method recovery was 85%-95%. Conclusion:The method is simple, sensitive, accurate and reproduci-ble. It is applicable in the pharmacokinetic study of fluvoxamine for clinical pharmacokinetics and bioequivalence studies.

Objective:To explore the repairing effects of 3D-printed nano-β-tricalcium phosphate (β-TCP) scaffolds loaded with vancomycin and bone morphogenetic protein-2 (BMP-2) for seawater -soaked tibial bone defects in rabbits. Methods:A total of 27 male New Zealand White rabbits were assigned to the normal group using a random number table method, with each group consisting of 9 rabbits. The rabbit tibial bone defect model was created using the osteotomy surgical method. Eight hours after operation, the wounds in the control group and seawater group were immersed in seawater for 2 hours, and those in the normal group were not immersed. After an observation period of 5-7 days, no significant redness or purulent discharge was observed in the wound appearance, then debridement was performed followed by corresponding implantations: the control group with gelatin sponges loaded with vancomycin and BMP-2, and the other two groups with 3D-printed nano-β-TCP scaffolds loaded with vancomycin and BMP-2. After filling the bone defects with the respective materials, all groups underwent layer-by-layer suturing of the wound, followed by disinfection with iodine and injection of gentamicin to prevent infection. The affected limbs were then immobilized using a plaster cast. The affected limbs were imaged using anteroposterior X-ray at 4, 8 and 16 weeks after operation, and the repair effects were evaluated using the Lane-Sandhu X-ray scoring system. At 16 weeks after operation, the bone defect tissues were collected for HE staining to observe bone tissue growth.Results:At 4 weeks after operation, the Lane-Sandhu X-ray score in the control group was significantly lower than that in the normal group [(2.8±1.1)points vs. (1.1±0.9)points] ( P<0.05), and that in the seawater group [(2.2±1.0)points] was not significantly different from those in the other two groups (all P>0.05). At 8 weeks after operation, the seawater group [(6.1±0.9)points] and the control group [(2.8±1.0)points] exhibited lower Lane-Sandhu X-ray score compared to the normal group [(8.2±1.0)points] (all P<0.05), and the seawater group showed a higher score compared to the control group ( P<0.05). At 16 weeks after operation, the control group [(3.8±1.0)points] exhibited a lower Lane-Sandhu X-ray score compared to the normal group [(10.0±1.3)points] and the seawater group [(9.3±1.2)points] (all P<0.05), while no significant difference was noted between the latter two ( P>0.05). At 16 weeks after operation, histological observations revealed varying degrees of bone tissue formation in three groups, with the normal group showing the best bone defect repair effect, followed by the seawater group. Conclusion:The 3D-printed nano-β-TCP scaffolds loaded with vancomycin and BMP-2 are effective for the treatment of seawater -soaked bone defects, which can promote bone tissue repair.

Objective To establish new classification criteria for early rheumatoid arthritis (E-RA) based on large samples of early inflammatory arthritis patients and to evaluate the value of this criteria in China.Methods Patients who had arthritic complaints with disease duration less than one year were enrolled.They were divided into RA group and non-RA group according to the clinical diagnosis by experienced rheumatologists.The clinical and laboratory parameters were analyzed and those with high sensitivity or specificity were selected as the new classification criteria.Statistical analysis was carried out by using t test,x2 test and Logistic regression.Results ① A total of 803 patients with early inflammatory arthritis were included in this study.Five hundreds and fourteen patients were diagnosed as early RA and 251 were diagnosed as other rheumatic diseases,and the diagnosis of 38 patients remained unestablished by the end of follow-up.② New E-RA classification criteria were established based on the parameters with high sensitivity and/or specificity.The sensitivity of the new E-RA criteria was 84.4％,which was higher than 1987 ACR criteria (58.0％),while the corresponding specificities were similar,which were 87.4％ and 93.6％ respectively.③ Compared with the complex scoring system of 2010 ACR/EULAR criteria,the E-RA criteria was more simple and practical.The diagnostic sensitivity and specificity of E-RA criteria were higher than those of 2010 ACR/EULAR criteria reported in the literatures.④ New classification criteria based on scoring system using Logistic regression analysis was established.The sensitivity of this criteria was 86.4％,which was higher than 1987 ACR criteria (58.0％).Conclusion The diagnostic value of the E-RA criteria developed in this study for early RA is better than 1987 ACR criteria,and is more simple than 2010 ACR/EULAR criteria.It may be used as a new classification criteria for early RA diagnosis.