Objective To prepare centromere-associated protein E(CENP-E)polyclonal antibody with specificity by using New Zealand white rabbits.Methods Prokaryotic expression plasmid of pHis-CENPEC410 was constructed by molecular cloning technique and then transformed into competent cells of E.coli BL-21 (DE3).HisCENPEC410 fusion protein was induced by isopropyl β-D-thiogalactoside (IPTG) and purified by affinity chromatography using Ni-NTA beads.The purified protein was used as antigen to immune New Zealand white rabbits to produce spccific polyclonal antibody of CENP-E.The antibodies serum was detected by immunoblotting and co-immunoprecipitation,and the purified antibodies were detected by immunofluorescene staining.Results The results of immunoblotting and co-immunoprecipitation demonstrated that the antibody serum was effective and the purified antibody could be applied to immunofluorescene test.Conclusions CENP-E polyclonal antibody with high specificity and sensitivity was obtained,which lay the foundation for the follow-up study of CENP-E.

Pharmaceutical care thinking is organic combination of the pharmaceutical and clinical thinking. Shang Han Lun contains very rich connotation of the clinical pharmacy, embodies the patient-centered and immediate intervention clinical pharmaceutical service concept in the progress of the diseases. This article sorted out and concluded pharmaceutical care thinking from the aspects of condition assessment and medication selection, medicine taking methods and notes after medication, which provided theoretical supports for the development of clinical pharmacy of TCM.

Objective To investigate effects and its mechanisms of hypertonic saline hydroxyethyl starch 200/0.5 solution on intracranial pressure and brain water content in rats with ischemic cerebral edema.Methods All experiments were conducted in the animal experimental center of Sun Yat-sen University.The 28 male Sprague-Dawle (SD) rats were randomly (random number) divided into hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group,control group and sham operation group,each n =7.Ischemic cerebral edema model was reproduced by middle cerebral artery occlusion (MCAO),followed by reperfusion after ischemia for 2 hours (If the moldel was not successful,other rats were operated to fill the missing models).Then reperfusion after ischemia 2 hours and received hypertonic saline hydroxyethyl starch and hydroxyethyl starch via tail vein at the beginning of reperfusion.The colloidal osmotic pressure (COP) and intracranial pressure (ICP) were evaluated on 0,2,6,12,18,24 hours after the surgery.The water content of the right hemisphere was measured on 24 h after the surgery.Results The ICP of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 2,6,12,18,24 h after the surgery.The ICP of hypertonic saline hydroxyethyl starch group was significantly lower than those of hydroxyethyl starch group and control group on 2,6,12,18 and 24 h.But there was no significant difference in ICP of the hydroxyethyl starch group compared with that of control group at all time points.The COP of hypertonic saline hydroxyethyl starch group and hydroxyethyl starch group were significantly higher than the control group and sham operation group at each time point; There was no significant difference in COP (mmHg) of the hydroxyethyl starch group compared with that of hypertonic saline hydroxyethyl starch group at all time points.The brain water content (BWC) of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 24 hours after the surgery [(81.24±0.36)％,(83.04±0.10)％,(83.14±0.41)％ vs.(78.37±0.37)％,all P=0.000],BWC of hypertonic saline hydroxyethyl starch group lower than these of hydroxyethyl starch group [(81.24±0.36)％ vs.(83.04 ±0.10) ％,P =0.000] and control group [(81.24 ±0.36)％ vs.(83.14 ±0.41) ％,P =0.000].There was no significant difference in BWC of the hydroxyethyl starch group compared with that of control group [(83.04 ± 0.10) ％ vs.(83.14 ± 0.41) ％,P =0.578].Conclusion Hypertonic saline hydroxyethyl starch solution could significantly ameliorate ischemic cerebral edema and reduce ICP,but the relationship between its elevated COP and reduced ICP has not been confirmed.

To enhance the capacity for extending and applying appropriate health technologies in rural areas in China,this paper proposes a supportive policy system that incoperates macro,average and microlevels. Thepolicysystemfocusesonorientation, incentives, regulationsand standardization,and its objectives and measures of each level are described.The policy system will contribute to the sustainable development rural health work.

Objective To observe the ultrasonographic features of local chest wall tumor recurrence after mastectomy for breast cancer and its clinical and histopathological characteristics. Methods The ultrasonographic features, clinical and histopathological characteristics of 27 patients with local chest wall tumor recurrence after mastectomy confirmed histopathologically were retrospectively reviewed. Results The disease-free intervals of twenty-seven patients ranged from 3 to 129 months [mean (31.9±31.4) months]. Most of the recurrence(18/27, 66.7%) occurred within 3 years after mastectomy. The clinical manifestations were:7 cases (7/27, 25.9%) with regional skin redness and swelling, red rash or ulceration on chest wall associated with or without palpable mass, 20 cases(20/27, 74.1%) with chest wall palpable masses without obvious skin change. On ultrasonography, 2 cases showed diffuse inifltrative type with ill-deifned inhomogeneous hypoechoic lesion and skin thickening. And twenty-ifve cases(43 lesions) showed mass type with a lesion size range of 5.4-114.7 mm [mean (24.4±21.6) mm]. Among them, 32 lesions were located near to the operation incision scar, 36 involved muscle layer, 38 were hypoechoic, 31 had irregular shape, 24 had indistinct margin, and 31 had blood lfow signal. In addition, calciifcation, halo, and taller-than-wide shape were absent in all 43 lesions. Conclusions The tumor recurrence often occurred within 3 years after mastectomy in high-risk patients. Ultrasonographic feature of chest wall recurrent lesion is of great value in the diagnosis.

Background Sclera remodeling process in axial elongation is one of the main pathological mechanisms of axial myopia progression.Studies confirmed that transforming growth factor-β1(TGF-β1) participates in the sclera remodeling process,and Smad3 is one of TGF-β1 downstream signal gene transcriptive factors,so to explore its role in sclera remodeling process of myopic eyes is of great significance for pathogenesis and prevention research of myopia.Objective This study was to investigate the expressions of type Ⅰ collagen and Smad3,a TGF-31 downstream target,in sclera of form deprivation myopic (FDM) eyes and explore the impact of TGF-β1-Smad3-type Ⅰ collagen signaling pathway on collagen remodeling in myopic sclera.Methods Seventy-five 1-week-old guinea pigs were randomly divided into normal control group (25 guinea pigs) and FDM group (50 guinea pigs).Monocular FDM was induced by occluding the left eyes of guinea pigs in FDM group with translucent latex balloons for 2,4,6 weeks,respectively,and consecutive occluding for 4 weeks followed by uncovering for 1 week (4/-1 weeks).The refractive power was detected by retinoscopy and axial length was measured with A-type ultrasound.Immunohistochemistry and reverse transcription-PCR were employed to detect the dynamic expressions of type Ⅰ collagen and Smad3 protein ad mRNA in the sclera of guinea pigs with emmetropia and experimental myopia,ard the relationship between collagen Ⅰ and Smad3 levels was analyzed.Results The refraction was hypermetropic in both normal control group and FDM group before occluding of eyes (P＞0.05),and the hypermetropic power was gradually reduced over time in the normal control group.In the FDM group,the refractive power was gradually changed from (+2.09 ± 0.31)D before occluding to (-1.23±0.69),(-4.17±0.59),(-7.07±0.56) and (-4.30±0.58)D,and the axial length was increased from (5.93-±0.39)mm to (6.62±0.36),(7.30±0.34),(7.99--0.32),and (7.21 ±0.36) mm at weeks 2,4,6,and 4/-1 after occluding,respectively,indicating significant differences in refractive power and axial length over time in the FDM group from normal control group and self-control group (all at P＜0.05).The expressions of Smad3 and type Ⅰ collagen protein and mRNA in the sclera of the FDM group was significantly lower than those of the control group and self-control group in various time points (all at P＜0.05).The positive correlation were found in the expression of Smad3 on the myopic sclera with that of type Ⅰ collagen in both protein and mRNA levels (protein:r=0.993,P＜0.05;mRNA:r=0.954,P＜0.05).Conclusions The myopic power and ocular axis increase dependent upon occluding time,and the expressions of Smad3 and type Ⅰ collagen in the sclera are correspondingly weakened in FDM eyes.A consistent expression trend is found between Smad3 and type Ⅰ collage,suggesting Smad3 and type Ⅰ collagen participate in the regulation of sclera remodeling in myopia by TGF-β1-Smad3-Collagen Ⅰ signaling pathway.

Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

Objective To construct the recombinant plasmid that highly expressed human subcellular PTEN,in order to provide a basis for further study on its anti-tumor effect. Methods PTEN cDNA was amplified by RT-PCR based on mRNA of placenta.The PCR product was ligated into T-vector,and transfected into E.coli;the obtained T-PTEN plasmid was identified with restrictive digestion and sequencing.PCR was used to incorporte nuclear signal of localization(NSL) into PTEN when T-PTEN was used as template.Then the PCR product was ligated into T-vector,and transfected into E.coli,and T-NSL-PTEN plasmid was obtained.pcDNA3.1 and T-NSL-PTEN were ligated after digested with EcoRⅠand BamHⅠ,and transfected into E.coli,the recombinant vector pcDNA3.1-NSL-PTEN was obtained,and identified with digestion and sequcncing.Results The recombinant expression vector DUM-PTEN and PUM-NSL PTEN were identified by restrictive digestion and DNA sequencing.As expected,by EcoRⅠ and BamHⅠ digestion,it showed the band of 1 200 bp.The sequencing result showed the NSL was incorporated successfully.The recombinant pcDNA3.1-PTEN was obtained with 1 200 bp,the sequencing result showed that its sequence was same as target gene;the recombinant pcDNA3.1-NSL-PTEN was comfirmed by restrictive digestion and sequencing,and the NSL was incorporated successfully. Conclusion The recombinant expression plasmid pcDNA3.1-NSL-PTEN is constructed successfully which can highly express human subcellular PTEN.

AIM To study the effect of Gardenia jasminoides Ellis on serum IL-1? and TNF-? level of rheumatoid arthritis rats, as well as its mechanism. METHODS An experimental type Ⅱ collagen-induced arthritic rats was established. The inhibitory effects on paw edema were observed, and serum IL-1? and TNF-? levels were determined in experimental rats. RESULTS Compared with models, Gardenia jasminoides Ellis inhibited paw edema on rats significantly, and the levels of serum IL-1? and TNF-? showed markedly decrease by Gardenia jasminoides Ellis at high dose to medium dose( P

BACKGROUND:Common therapies for damage to the central tendon of finger extensor tendon include Matev, Carrol, Fowler methods as wel as residual central tendon flipping repair, but the therapeutic effects are no satisfied with bloated appearance, tendon adhesions, limited joint function. OBJECTIVE:To investigate the clinical effects of dorsal longitudinal driling and segmental tendon graft to repair old central tendon injury. METHODS:Eighty patients with old central tendon injury were randomized into treatment group and control group, with 40 cases in each group. In the treatment group, dorsal longitudinal driling and segmental tendon transplantation were given; while in the control group, Carrol, Matev, Fowler methods were chosen according to the individual conditions. Then, the therapeutic outcomes were compared between two groups. RESULTS AND CONCLUSION:The excelent and good rate was 85% in the treatment group and 65% in the control group, and there was a significant difference between the two groups (P < 0.05). Results from Valpar Component Work Samples showed that the number of cases adapting to the original work was 30 cases (75%) in the treatment group and 16 (40%) in the control group, and there was also a significant difference between the two groups (P < 0.05). The degree of proximal interphalangeal joint flexion was increased gradualy in the two groups at admission, at 15 days after hospitalization, at 1 day before discharge and at 4 months after discharge, and meanwhile, the degree of proximal interphalangeal joint dorsiflexion was reduced gradualy (P < 0.05). There were significant differences in the degree of proximal interphalangeal joint flexion and degree of proximal interphalangeal joint dorsiflexion between the two groups at 15 days after hospitalization, 1 day before discharge, and 4 months after discharge (P < 0.05). These findings indicate that the dorsal longitudinal driling and segmental tendon graft for repair of old central tendon injury can play an effective role in the recovery of articular flexion and extension function.